"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

For those absolutely devoid of scruples, charity fraud is the field par excellance, in which you can simultaneously harvest kudos for your humanitarianism and make off with vast bundles of untaxed cash. Convictions for charity fraud are so rare as to be nonexistent, so any criminals operating in other fields of endeavor are incurring unnecessary risks.

"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:22 am

"GOOF" COSTS 65 YEARS TO INFINITY OF CANCER RESEARCH.
Excerpt From: "A Conspiracy of Cells: One Woman's Immortal Legacy and the Medical Scandal It Caused"
by Michael Gold
© 1986 Michael Gold

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Henrietta Lacks

1: Special Delivery

Jim Duff was carrying an odd little suitcase as he stepped out of the helicopter. It was a box of molded styrofoam, like a small ice chest you might take to a softball game to keep a six-pack cool. Except that this chest was made of thicker, more serious looking stuff.

Walter Nelson-Rees knew what was inside. It was the reason he had come out to the Berkeley heliport to meet Duff. The two men shook hands, and Nelson-Rees noticed that Duff was a bit edgy.

"We'd better go get a drink," said Duff.

When they had driven the few blocks to Oakland and found a bar, Duff gingerly placed the box onto the floor beneath their table and started his pitch. "Walter," he said in a quiet, earnest tone, "these cells are ... different. And you're going to have to handle them ... differently."

Of course the cells are different, thought Nelson- Rees. When had a bureaucrat from the National Cancer Institute ever flown from Washington, D.C., to California to personally deliver a chest full of dry ice and malignant tumor cells? There had been plenty of shipments over the years-hundreds, in fact-but air freight had always sufficed. The National Cancer Institute trusted the airlines, and when the airlines got the packages to Oakland, it trusted Nelson-Rees to handle the rest.

Tall, with a sharp nose and thin lips, Walter Anthony Nelson-Rees had a slightly aristocratic look about him and a manner that suggested he was sure of everything he did. He operated one of the best cell banks in the country, if not the best, and seemed quite aware of that. The officials of the National Cancer Institute knew it too. In fact, Nelson-Rees's cell bank existed largely to serve researchers working under the aegis of the institute. They were the bigshots, some of the best scientific minds in the country, and they needed the best weapons they could get to fight "The War."

"The War on Cancer," as the newspapers called it. The year was 1973. Richard Nixon wanted to be remembered as the president who brought cancer to its knees, and the National Cancer Institute, lavishly endowed, was his war department.

Yes, the institute wanted only the best for its boys, the choicest cuts of cancerous tissues from which they might extract the secrets of the disease, its cause and its cure. Nelson-Rees was the supply man. Cells from any organ taken from patients of any age, any race, either sex -- you name it, he had it. When a researcher managed to get a new kind of cell growing in a culture dish, a cell that looked as though it could be useful for cancer experiments, chances are the institute would have a sample sent to Nelson-Rees.

He was as fastidious as The Odd Couple's Felix Unger, which was the perfect recommendation for someone in charge of nurturing sometimes delicate cultures, keeping their identities straight, and protecting them from ... well, from anything unexpected. Nelson-Rees complained regularly to maintenance workers at his lab about their inability to get the air pristine, free of dust and microorganisms. And few of those who saw it will ever forget the scene following Nelson-Rees's discovery that a technician had used a hallway in the "clean section" of the cell bank to fold up his parachute after hours. "His spore-infested parachute!" wailed Nelson-Rees.

That's why he was one of the trusted keepers of the cells. He was a perfectionist.

Yet here was Duff, straight from a transcontinental flight with his cancerous carryon luggage, going on about how special these latest cells were and how careful Nelson- Rees was to be. "You're not to do anything with these, Walter, unless you get a prior okay from us. Just thaw them out, grow up a few separate populations, and store them away." Jim Duff was the man who watched over Nelson-Rees's lab for the institute. He was a natural bureaucrat with a bureaucrat's instinct to cover his rear flank. But he was also a friend to Nelson-Rees, who therefore nodded politely as he sipped his vodka and tonic.

"This is very important, Walter. There's more to it than just science." Duff paused to let the message sink in.

Inside the styrofoam case, within sealed plastic flasks, floating in a red nutrient bath at room temperature, some of the cells were growing and multiplying as the two men spoke. In a separate refrigerated compartment, duplicates of the active cells sat motionless in tiny glass bottles, suspended in a frozen state. There were six distinct cultures in all, a six-pack of human tumor cells, six new weapons for "The War."

And Duff was right; they were different. For one thing, they had come from the Soviet Union.

Richard Nixon not only dreamed of curing cancer, he also hoped to take the chill off America's relationship with the Russians. Toward both ends he negotiated an agreement with Leonid Brezhnev in May of 1972 that called for the two nations to cooperate in biomedical research. Soviet scientists were keenly interested in "The War," particularly any recent progress that their American counterparts, who were thought to be at least several years ahead, were willing to share. So in November the first delegation of American cancer researchers traveled to Moscow to present their Russian colleagues with, among other things, a set of thirty viruses. There were rat viruses and hamster viruses and wooly monkey viruses and gibbon ape viruses, all of which had been found to induce cancer in these animals.

By studying viruses that caused cancer in animals, researchers hoped to find out something about analogous viruses that might cause human cancer. In fact in the early 1970s, almost everyone who counted at the National Cancer Institute seemed certain that in only a few years someone on their payroll would find the virus responsible for cancer in human beings. So far, though, all they had turned up were viruses that produced cancer in animal cells. Even those few isolated from human cells turned out to be animal viruses that had somehow found their way into human tissue but were incapable of triggering cancer there.

Undaunted, the institute pushed ahead, spending $60 million a year in the search for the human tumor virus. Competition was stiff, to say the least. For to find the virus that causes human cancer would open the way to a vaccine, a shot, like the miracle vaccination that had been immunizing people against the plague of polio for the last few decades. Cancer would be on its way to extinction, and a Nobel Prize would be the least an appreciative world could do for the scientist who made it possible. The high-pressure race had already led to several premature claims of victory in the United States. One of the most recent "winners" of the Human Cancer Virus Sweepstakes even received official congratulations from Richard Nixon after the good news was leaked to the press. Some months later the alleged agent was determined to be another animal virus -- this one a mouse virus -- and the race was on again.

In any case, the National Cancer Institute figured it wouldn't hurt to give the Russians a few of these animal viruses. The collection was all done up in handsome gold lettering and presented with much pomp. It was such a hit among the Russian scientists that they felt they had to give the Americans something special in return. What was special about the cultures of human cancer cells delivered by the Russians a few weeks later was that all six had viruses growing within them, viruses that the Soviets suspected were the causes of the malignancies.

Well, as they say in diplomatic circles, it was quite something. Richard Nixon's scientific delegates were unanimously skeptical that the Russians, with their primitive equipment and lax laboratory techniques, could have come up with even one cell line that contained a genuine human cancer virus, much less six of them. No, it was too much to believe. And yet there they were, six cultures of cancer cells taken from six different patients, each culture carrying some kind of virus. Quite something. "My God, suppose they somehow stumbled onto it?" the Americans asked each other. It was simply too potentially valuable to ignore. Besides, even the purest of cynics on the American team knew there was more at stake here. These were Russians, this was detente, there were political ramifications. The Americans smiled and thanked the Russians. They packed the cells into their suitcases and brought them back home.

A couple of weeks later, Walter Nelson-Rees received a letter from the institute saying that samples of some virus-laden Russian cells, received in connection with the biomedical exchange program, would soon be delivered to his cell bank. Only four other American scientists had been given samples of the cells, and only they were authorized to experiment with them, the letter explained. As for Nelson-Rees, he was to keep some of the cells growing and to freeze the others as a safeguard against loss. The letter concluded: "No one is to be provided with these materials, or with any data acquired using these cultures, unless specific authorization has been obtained .... Dr. James Duff expects to visit you on January 10, 1973. He will hand carry the cultures to insure safe delivery."

Nelson-Rees thought it improbable that the viruses in these cells would prove to be human cancer agents. He was, however, pleased with himself and his lab. Once again the bigshots had come to his tidy cloakroom to check their fanciest hats.

***

When they had drained their drinks, Duff picked up the Styrofoam case and followed Nelson-Rees to the car. It was about nine o'clock in the evening. They drove through Oakland's harbor district to the Naval Biosciences compound. It was here, through a confusing web of agreements between government agencies and the School of Public Health of the University of California at Berkeley, that Nelson-Rees operated his cell bank. Before they entered the clean section where the cell culture work was done, where floors as well as counter tops gleamed like mirrors, Nelson-Rees asked Duff to remove his jacket and don a white lab coat. Then he handed him a pair of white nylon booties to keep the street dust on Duff's shoes from dirtying the floors.

Nelson-Rees recorded the arrival of the cultures in a big black logbook. He put the plastic flasks in a tall aluminum cabinet that looked like an oversized refrigerator but in fact was the incubator, where cells in culture grew in a tropical environment of 98.6 degrees Fahrenheit. The tiny glass bottles went into a cylindrical freezer of stainless steel -- about four feet in diameter and five feet tall -- that was filled with liquid nitrogen at 300 degrees below zero. Nelson-Rees slammed the lid shut, forcing a cloud of cold, white vapor into the room, and turned to Duff.

"Okay?"

"Okay."

Walter Nelson-Rees's obsessions went far beyond nylon booties. "In order to run a successful cell bank that stores thousands of different tissue cultures, that grows up and sends out as many as a dozen samples a day to researchers all over the country -- to do that," Nelson-Rees often explained, "one needs to know what one is working with. One has to know which cell is which." Identification, the art and science of being able to recognize the subtle differences between cells, was his stock-in-trade. All of which is to say that he could no more leave six unidentified cell cultures in his deep freeze and incubator than Felix Unger could sneeze into his hands.

A few weeks after Jim Duff told him not to do anything with the Russian cells, Nelson-Rees disobeyed orders. He sent samples of each culture to Ward Peterson, a colleague at the Child Research Center in Detroit, who specialized in performing certain biochemical tests. This was always the first step in his routine check of anything that came into the bank. While waiting for Peterson's results, Nelson-Rees examined the cells' chromosomes, the rodlike structures inside each cell's nucleus that carryall its genetic information. After several days of gazing through a microscope at scores of slides, he had convinced himself that indeed, these cells carried human chromosomes; they were human cells. If someone had said to him at that point, "Well of course the chromosomes are human. The Russians said they were all human cells," Nelson-Rees would have nodded and replied, "Mmm, of course. But one must be sure about these things."

The cells were from human patients, all right, but he had noticed something odd. Among its many chromosomes, every human cell usually has two that determine an individual's sex. The sex chromosomes come in two varieties, one shaped roughly like the letter X, the other like a Y. If a person's cells have two X chromosomes apiece, that person is a female. If they carry one X and one Y, the person is a male. What Nelson -Rees noticed was that there were no Y chromosomes among any of the Russian cells. How strange if by chance all six cultures happened to come from women. Of course, it was well known to tissue culture experts that cells growing for a long time in a laboratory environment could simply lose chromosomes now and then. Indeed, the Y seemed particularly vulnerable to getting lost in the shuffle of division. Possibly there were some males among the original Soviet cultures whose Y chromosomes had been lost along the way; that would explain things.

Still, it bothered him.

Peterson's results began coming in about a month later. He had been analyzing the cells' enzymes, catalysts that speed up chemical reactions within the body. While some enzymes are standard equipment for human beings -- nearly everyone, for instance, has the enzyme galactose-I-phosphate uridyl transferase that helps break down a sugar found in milk -- certain ones appear only in small segments of the human population. These enzymes, once observed and catalogued, can be used along with other characteristics to tell cells apart.

In the first Russian cell line he tested, Peterson found an enzyme called G6PD type A, a variant of a standard enzyme that helps to metabolize glucose, another basic sugar. The form of G6PD in most people moves relatively slowly through an "electrophoretic gel," a kind of Jell-O smear hooked up to a battery or transformer. Electrophoresis indirectly measures a chemical's electric charge by testing how fast a certain voltage can drag it through this gel. The point is that in addition to the common, slow-moving form of G6PD, there is another variety that moves faster, presumably because it has a stronger charge. It was known from studies of many cells that this variant, called type A, occurs almost exclusively in black people. And even among blacks it appears in just one out of three people. Peterson's discovery of this uncommon enzyme in one of the Russian cells was not in itself unusual; there are blacks in the Soviet Union. What was unusual was his finding it in the second cell line as well, and, as the months went on, in the third, fourth, fifth, and sixth.

"It is somewhat unexpected that all six cultures would lack the male Y chromosome," Nelson-Rees said to himself. "It is even more unlikely that all six cell cultures would by chance be from blacks. And the odds that all would also just happen to have the rare form of the G6PD enzyme are .... Well, it's almost impossible."

He decided to notify the bureaucrats in Washington.

Jim Duff, long-distance, was upset, though not as much as might have been forecast based on his original commandment on the special handling required for these cultures. Perhaps he knew Nelson-Rees well enough to expect his friend would scrutinize them and send them to Detroit despite the warnings. Or maybe the news left him stunned. In any case, he told Nelson-Rees the observations were unusual, but they pointed to no firm conclusion. No doubt it occurred to Duff that, if true, the findings might eventually prove to be embarrassing. He was inclined not to accept them, at least not until Nelson-Rees had checked and double-checked. "There's too much at stake here, Walter," said Duff. "Don't rock the boat. Don't rock the boat until you're absolutely, 100 percent sure."

The next call went to the man at the institute who had direct responsibility for the care of the Russian cells. He was a superior of Duff's, superior not only in rank but also in his ability to worry about matters of protocol, to evaluate every development in terms of how it might offend someone, anyone, or -- who knows? -- maybe even endanger his job.

"What do you mean you sent the cells out for testing?"

"I sent them out for the purpose of identifying them," said Walter Nelson-Rees.

"Those cells were not to go anywhere. What has this fellow in Detroit been doing with them?"

"Nothing, just analyzing them. You needn't worry." But the bureaucrat was worried, and one of his more intriguing worries was that the institute might get nailed for violating federal quarantine procedures. It was awkward enough that Nixon's delegation had smuggled the Russian cultures into the United States in the first place. Being bathed partly in bovine serum, the cells were a potential source of hoof-and-mouth disease, a scourge then rampant in the Soviet Union. Authorities with the U.S. Department of Agriculture had become quite agitated when they heard about the cells' unofficial entrance into the country. They had calmed down slightly when the cancer officials permitted inspections of the few labs authorized to work on the cultures. But now, if news leaked out that Nelson-Rees had been shipping the stuff wherever he pleased ....

"Doctor Nelson-Rees, you were told specifically not to distribute those cells. How could you have sent them to Detroit?"

"I've just told you. I sent them to Detroit to be identified."

By the time the conversation had circled back around itself a few times, it was clear to Nelson-Rees that the bureaucrat was not interested in hearing about his findings. He then called Wade Parks, an institute virologist and one of the privileged researchers allowed to examine samples of the Russian cells. Parks said he had tentatively identified the virus from one of the cell lines. Unfortunately, it appeared to be a monkey virus -- another confounded animal virus. This one, Parks said, had never been known to do anything related to cancer or to be anywhere interesting. Not only that, preliminary checks of the other cell cultures suggested they might all be carrying the same worthless virus. The implication was that the Russian cells were duds. The elusive human tumor virus, it appeared, remained elusive.

But Nelson-Rees was more interested in the cells themselves than the viruses they held. What with his own findings and now this from Parks, there was something obviously strange about these cell lines. All lacked the Y chromosome, all carried the rare enzyme, and now all contained the same virus. The theory taking shape in his mind was both obvious and preposterous; he had to try it out on someone. "Has it ever occurred to anyone that these might be all the same cells?" he asked the virologist.

He would not have been surprised to hear Parks dismiss the idea. After all, how could they be the same cells? Each culture had come from a different research institute in the Soviet Union. Each was seeded by a bit of tissue taken from different patients with different kinds of cancer. How could they now be identical? And what kind of cell would they all be? Parks, however, said none of this. Instead he said that the same thought had occurred to him.

Nelson-Rees couldn't say -- not precisely, anyway -- how all the cell lines had become one. Nor could he say what kinds of cells were in the six cultures originally. But he was almost sure of what was growing in them now. He even knew the name of the woman to whom the cells, all of them, belonged. But he needed proof.

As it happened he and an assistant had just learned a technique that could help him get the proof. It was a method of applying purple stain to a cell's chromosomes. The chromosomes absorbed the color in selected areas and ended up looking like barber poles. Instead of neat stripes of red and white, though, they had bands of purple running across them at irregular intervals. The banding patterns were unique to each chromosome, like fingerprints, which made cells eminently more identifiable. If a researcher knew the banded fingerprints for various cell lines, he could use them to identify an unknown cell with far more certainty than by merely determining the lack of a Y chromosome, for example, or the presence of a rare enzyme.

Nelson-Rees and his assistant spent several months preparing the Russian cells, staining the chromosomes, photographing the fingerprints, and checking them against the fingerprints of their prime suspect. When they were through, the conclusion was inescapable. Maybe the bureaucrats didn't want to hear it, but they got another phone call. It was bad news, reinforcing Parks's impression that the cells were probably worthless to the quest for the human tumor virus. But it went beyond that.

These were not distinct cultures of cancer cells from six different patients in the Soviet Union. They were all the cells of an American who in her entire life had probably not been more than a few miles from her home in Baltimore, Maryland.

A housewife with four children, this woman had been stricken with cancer at the age of thirty. She died in 1951, more than twenty years before a group of smiling Russians proudly presented a group of smiling Americans with six of their most promising cell cultures.

Her name was Henrietta Lacks.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:23 am

2: The Seed That Took

Who? According to the medical records, Henrietta Lacks was born somewhere in Virginia on 18 August 1920, the daughter of John and Eliza Pleasant. Her mother died while delivering her tenth child; Henrietta was very young at the time. Under the section titled "Personal and Social History," the records say that Henrietta was the mother of four and that she had a happy home life. Her major anxiety was due to a ten-year-old epileptic daughter who could not speak. As for education, Henrietta had gone as far as seventh grade. She drank beer only occasionally. Her husband David worked at the Sparrow's Point shipyards and earned an adequate income. They rented a five-room house with coal heating on New Pittsburgh Avenue in Baltimore.

Under "Past Health" the description is medically unremarkable: As a child she had chicken pox, measles, and periodic trouble with her tonsils. As an adult she had occasional headaches and often suffered a stuffed-up nose, probably the result of a deviated septum. In short, the records describe a young woman who had been quite well and relatively free of cares until January of 1951, when she noticed a pink discharge spotting her underclothes.

By 1 February, the day Henrietta Lacks made her way through the winter rain to the women's clinic of the Johns Hopkins Hospital, the discharge was blood red.

The gynecologist who examined her was baffled. He quickly found the source of the blood; it was a puffy lobe of tissue about an inch in diameter on the left side of her cervix. But as to what it was exactly, this experienced physician couldn't say. All the cervical tumors he had encountered were pale, almost white, because they lacked an ample blood supply. In fact most were so starved for blood that they developed ulcers. Not this thing; it was shot through with vessels and so full of blood that it seemed to glow a deep red. There were no signs of ulceration either. It was a mean and hardy thing, whatever it was.

The gynecologist took Henrietta Lacks across the hallway to the venereal disease clinic, thinking it might be a lesion caused by syphilis. But the technicians there could find no trace of syphilis bacteria within her cervix. Still puzzled, the gynecologist cut off a small section of the lump and asked a pathologist to examine it under a microscope. Henrietta Lacks received the verdict that same day: an unusual form perhaps, but this dark red growth of tissue was unquestionably cervical carcinoma, a malignant tumor of the cervix.

Eight days later she returned to the hospital for her first session of radiation therapy. A surgeon stitched a tube containing radium capsules to the wall of her cervix. It remained there for twenty-four hours, bombarding the tumor and the rest of her abdomen with high-energy gamma rays. After a second operation to remove the tube, the surgeon wrote his report: "Patient feels quite well tonight. Morale is good and she is ready to go home." His records also note that during the earlier operation, he had sliced off two tiny pieces of cervical tissue -- one normal, the other from the tumor -- and had given them to a researcher in the Hopkins medical school named George Gey.

George and Margaret Gey were husband-and-wife pioneers in the field of tissue culture. George, who stood well over six feet tall and had a broad chest, was an idea man and a tinkerer. Among his many inventions was something called a roller tube, a sealed glass vessel in which living cells were periodically bathed in a nutrient solution as the tube was slowly rotated. He blew the glass for the first tube himself and rigged up a primitive rotor using the pendulum of a clock. It became a standard in the field. A zealot when it came to the care and feeding of cells, George Gey once bought an entire boxcar full of powdered soap for washing culture dishes; it seems the local market had switched to a newfangled detergent that sometimes left residues harmful to cells, and George wanted to make sure he'd never be forced to use the stuff. Although a native of Pittsburgh, he had a country boy's air. He spoke plainly and with warmth as readily to cab drivers as to scientific dignitaries. He ran his lab like an informal college of tissue culture, sharing equipment and knowledge with any student or scientist who wanted to learn. And for a few hours every Wednesday, he went fishing.

Margaret, trained as a surgical nurse, was the meticulous director of day-to-day operations in the laboratory. As long as everyone did his share, she was an amicable boss. If necessary, however, she could play the role of the stern head nurse. She saw to it that the cultures were fed on schedule, that the glassware was sterilized according to specifications, that the records were kept in order. George was regularly out of the lab, hunting for funds and lecturing, but Margaret was always there, working long days and weekends, all of it without pay.

Together the Geys were trying to put some science into the folk art of growing cells. If reliable methods could be devised to keep cells alive in a roller tube and to trick them into thinking they were still inside a body, then scientists could learn firsthand about human biology without having to experiment on human beings directly. The workings not only of healthy cells but also of sick ones could be revealed. That possibility -- of capturing human diseases under glass and dissecting them for their underlying causes -- was what lured the Geys and several other research teams into the business of culturing tissue. Their greatest hope was to establish and study long-lasting cultures of the most dread human disease, to have, as some put it, "a tumor in a test tube."

But it was slow, difficult, sometimes grizzly work. In order to give their cells a proper feeding solution, for example, they had to find the raw materials themselves. Several times a week Margaret went to the Hopkins hospital's maternity ward to collect one ingredient believed to stimulate cell growth: blood from human placentas. One of the maternity nurses would press a button that set off a buzzer in the Geys' lab, signaling that a fresh placenta was being put aside for them. Margaret soon arrived, cleaned off the sac's umbilical cord, and, plunging a fat syringe into one of the cord's larger blood vessels, pulled out as much as 50 cubic centimeters, about a third of a cup. Back at the lab the serum component, the slightly yellowish fluid containing proteins, was removed from the blood and combined with what the Geys called beef embryo extract. This was the ground-up remains of a three-week-old cattle embryo, collected periodically from a cooperative packing house. Because the recipe also called for chicken plasma, every so often the Geys visited a nearby poultry factory. They usually went about dawn so that few workers would be around to witness the spectacle. Margaret's job was to pull back the wing, swab the area around the ribs with alcohol, and hold the animal still while George poked the syringe directly into its heart. They took 50 cubic centimeters from each chicken, and most walked right off the table and back to the yard. They had a deal with the owner to buy any bird that didn't survive. On such occasions Margaret cooked the unlucky animal for dinner that evening.

As for the cells to be grown in this elixir of plasma and serum and embryo mash, the Geys had to go out and collect them as well. Every day George scanned the hospital's list of upcoming operations and procedures, looking for interesting sources of tissue. He or Margaret or one of the technicians stood in the appropriate operating room holding a few petri dishes ready for a scrap. Then they raced back to the lab, placed the pulpy fragment on a clot of chicken plasma, added the remaining parts of their cell food, and hoped that it would take.

It was discouraging. No matter how clean their glassware, no matter how potent their nutrient solution, no matter how careful their technique, cells simply weren't comfortable growing outside the human body. The Geys had had considerably more success with animal cells, including some that had survived for years now. But most human cells shriveled and died right away. Some held on for a few weeks, coaxed and coddled by Margaret's diligence, and then gave up. It was a testimony to the Geys' abilities that by early 1951, they had managed to keep a few lines of human cancer cells alive for several months. It was a testimony to their determination that they kept trying to establish new ones.

Mary Kubicek, however, was running low on determination. A twenty-one-year-old technician just out of college and newly trained by the Geys, Mary was frustrated by her current assignment: an attempt to establish a culture of cervical cancer cells. Mary was shy and slightly insecure; that made her doubly careful and especially hardworking, even measured by the high standards of the Gey lab. Nonetheless after trying samples of cervical cancer from a dozen different patients, all she and her fellow technicians had to show were dead cultures. When George Gey announced around noon on 9 February that he had dropped off yet another cervical cancer biopsy in Mary's work area, she didn't attend to it right away. Uncharacteristically, she lingered a few minutes at the lunch table and finished her sandwich. What's the difference, she thought. It'll just be another useless attempt.

The tumor fragment was red, roughly square, less than half an inch on a side. Gey explained that it had been taken from a patient in the women's clinic just before she underwent radiation therapy. Following the usual procedure, the tissue was code-named with the first two letters of the donor's first and last names: HeLa. Mary cut away the decaying, brown tissue around the edges and sliced the remaining chunk into tiny cubes. She pipetted a few drops of chicken plasma into several roller tubes and placed four cubes of tissue in each tube. She waited five minutes for the plasma to stiffen into a clot, and then added the rest of the feeding solution. Finally, she placed the tubes into a rolling rack inside the laboratory's incubator.

Had Mary been staring through the glass doors of the incubator, she wouldn't have seen the early signs of life. It happens so slowly at first and so sporadically. Maybe it was a matter of hours, maybe the better part of a day passed. At some point, the cells in each little island of tumor began to quiver and dance ... and multiply. Where there had been one, there were now two. Where there had been two, now four, now sixteen, now thirty-two. In a few days, the signs of growth were visible: around each cube a translucent band of new cells was taking shape. On the fourth day Mary had to remove the burgeoning bits of flesh from the roller tubes, carve them up into smaller pieces, and transfer the cuttings into additional tubes.

Every other human cell line in the Gey lab had eventually weakened and faltered. Yet as the months passed the HeLa cells showed no such vulnerability. They just kept growing, doubling their number every twenty-four hours "spreading like crabgrass!" was how Margaret Gey described it. At one point George Gey compared the performance of the cancerous HeLa cells to the performance of the normal cervical tissue taken from Henrietta Lacks: the tumor cells were growing ten to twenty times faster. It was too early to start celebrating, and George Gey wasn't the sort to stop and congratulate himself anyway. But there was no doubt about it, these cells were different.

***

The tumor within Henrietta Lacks was different too, though the doctors didn't know it at the time. Most cervical tumors -- especially those found at an early stage, as was Henrietta's -- were easily beaten back by radiation. Most patients were still alive five years after therapy. So after several more radium treatments, the doctors gave her one month of X-ray therapy and hoped that would be the end of it. "No symptoms referable to the pelvis," wrote one who examined her in May. "Cervix is normal in size, mucosa red and smooth, cervix freely movable. Good radiation result. Rx: Return in one month." In June he wrote: "Patient feels fairly well, but continues to complain of vague lower abdominal discomfort. Cervix appears perfectly normal. No evidence of recurrence. Rx: Return in one month."

By late July Henrietta's side aches could no longer be described as vague. Now they were extreme and radiating down into the groin. Not only that, something was constricting the area around her bladder so severely that her kidneys began to swell with urine. The doctors also found a large, stony mass of tissue on the inside of her pelvis and another tumor in a lymph gland. With shocking speed the cancer had reappeared and spread so extensively that the doctors considered it inoperable: "For this reason, we are giving the patient a second course of deep X-ray therapy purely as a symptomatic therapy."

She was admitted to the hospital on 8 August, ten days before her thirty-first birthday. For the next twenty-two days she ran a constant fever of 100 to 102 degrees; she also began vomiting regularly. Despite the X-ray treatment, tumors were filling her abdomen. The treatment was halted.

In mid-September, with the fever, pain, and nausea continuing, she developed uremia, a buildup in the blood of poisonous waste products normally eliminated by the kidneys and bladder. The doctors tried to insert a catheter tube through the mass of tumors that choked her bladder, but failed. They gave her transfusions, replacing her own polluted blood with a fresh supply. Her intestine, however, was also blocked and her abdomen was beginning to expand.

On 26 September one of the doctors looked over her order sheet, the long list of various drugs, procedures, and treatments that had been prescribed to try to save Henrietta Lacks. At the bottom of the list, he scrawled: "Discontinue all medication and treatments except analgesics." All he could do was try to relieve the pain. On 29 September, Henrietta Lacks became disoriented, apparently confused about where she was and what she had been going through. She stopped breathing at fifteen minutes after midnight on 4 October 1951.

It had been only eight months from the time the small red patch was first discovered in her cervix to the day it killed her. For cancer of the cervix, the doctors said, it was some kind of a record.

***

No one in the Gey laboratory laid eyes on their unfortunate benefactor until 4 October when George Gey noticed a listing for the autopsy and went to observe. Because he wanted a few more samples of the remarkable tumor cell, he asked Mary Kubicek to meet him there and collect them.

The Hopkins autopsy room was buried in the basement of another building. Mary had never been there before. In fact she had never been to any autopsy facility or to a morgue or to anything like that. She made her way anxiously through the maze of dim underground hallways that connected the laboratory building to the other basement.

The room had high ceilings and a bare stone floor. At the far end Mary saw a body on the table. A pathologist was hunched over it, at work, and Gey stood nearby. The dead woman's arms had been pulled up and back so that the pathologist could get at her chest. Even from a distance Mary could see that the body had been split down the middle and opened wide.

She walked to the table, sidestepped one of the outstretched arms, and held out her petri dishes. As she waited she gaped at the greyish white tumor globules that filled the corpse. It looked as if the inside of the body was studded with pearls. Strings of them ran over the surfaces of the liver, diaphragm, intestine, appendix, rectum, and heart. Thick clusters were heaped on top of the ovaries and fallopian tubes. The bladder area was the worst, covered by a solid mass of cancerous tissue. "Bladder pushed to anterior abdominal wall," wrote the pathologist, "Almost entirely replaced with tumor."

Mary's eyes wandered down toward the corpse's feet and suddenly she was overcome. The toes. They were painted with bright red nail polish, and a dainty job it was. It suddenly made this carved-up cadaver real. All the laboratory experimentation had never hinted at the tragedy of this disease. But here, she thought, over here on the table is the proper demonstration. Here is what cancer does.

The pathologist sliced pieces of tumor from different organs and dropped them one-by-one into the dishes in Mary's hands. It seemed to Mary that he took forever. Finally, she fled: across the expansive stone floor and out of the autopsy room, through the catacombs, and up the stairs from the basement. Back at the lab she concentrated on the task at hand, cutting up the tissue and placing the pieces into roller tubes. In the bright familiar surroundings, her horror quickly faded. The image of the delicately polished toenails, however, lingered. In the years that followed, that sight came back to her often.

As for the cadaverous cells, they would not grow. The uremia had made it impossible for anything to live within the body of Henrietta Lacks. The cancer's sabotage had been so effective, it killed not only its host, but also itself.

That's not quite right, of course. Part of what had been Henrietta Lacks's cancer was not dead. Some of the cells had escaped their own poisonous wreckage eight months earlier on the edge of a surgeon's knife, abetted by the tissue culturing skills of Mary Kubicek. Dining on clotted chicken plasma, chopped beef embryo, and the blood from human placentas, the surviving cancer cells of Henrietta Lacks were living quite comfortably -- thriving, in fact -- in glass tubes in George Gey's lab.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:23 am

3: HeLagram

In the early 1950s many Americans would have named cellophane science's niftiest invention. To the small community of researchers trying to grow human cells in their laboratories, however, the truly great breakthrough was the HeLa cell. At last, here was a cell with staying power, the first durable piece of a human being that could be watched up close and tinkered with. No more racing to finish an experiment before a sickly culture wheezed and fell dead. This cell would last, not just through one series of tests but through dozens, for months, maybe for years.

In fact George Gey and two co-workers at the University of Minnesota showed that HeLa cells were so rugged they could survive a 2,500-mile trip through the mail. They sent twenty-nine live cultures by air, rail, and truck from Minneapolis to Norwich, New York, and back; all but one returned in fine health. A cell line that held on in the lab was what everyone had been hoping for, of course, but one that could endure handling by the U.S. Post Office was a blessed miracle.

Soon it seemed every biomedical scientist in the country was either sending or receiving a HeLagram. Gey started it by mailing samples of HeLa cells to a few close colleagues, who grew up some extra cultures and sent them to their friends, who did likewise. When the frenzied demand for HeLa outpaced this informal network, a number of laboratories set up full-scale production lines and began passing around HeLa cultures the way McDonald's shovels out its burgers and fries. Mary Kubicek heard that one shipment of HeLa was being carried by backpack into Chile and another was on its way to Turkey. In a few years HeLa cells would even travel into space aboard the Discoverer XVII satellite.

Cancer researchers craved HeLa most of all because it was their long-sought tumor in a test tube. They watched the cells react to a battery of toxic chemicals and photographed the weirdly shaped chromosomes. Scientists interested in the general workings of human cells clocked the rate at which HeLa cells multiplied and studied their production of proteins. Virologists, who found that polio viruses multiplied a millionfold just two to three days after infecting a HeLa culture, made HeLa their major new tool for studying the viruses. A year later they knew enough about polio to produce the first successful vaccine.

And, of course, every scientist who wanted to learn the methods of tissue culture insisted on starting with HeLa. The newcomers had been told how frustrating much of the work would be, how rarely a piece of tissue would yield a sustainable cell line. But with HeLa, they knew they couldn't lose. The advent of HeLa was not only a boost for the beginners, though. It also seemed to change the luck of the researchers who were struggling to establish other human cell lines.

It was as though George Gey had broken biology's four-minute mile. Once he had shown it was possible to produce a strong and long-lived cell line from human tissue, his fellow cell culturists went back to the lab with new confidence and enthusiasm -- and damned if they weren't able to do it too. One scientist successfully cultivated a hardy strain of human liver cells. Another started up a cell line from a bit of amniotic sac. Then came a culture from a tumor of the larynx, followed quickly by sturdy lines of embryonic kidney cells and adult heart cells and cancerous blood cells. True, the cultures didn't always bloom right away. But eventually, a few days or a few weeks after being seeded, somehow they all began growing at a strong and steady pace. For the next ten years, from the early 1950s to the early 1960s, the science of cell culture flourished as well.

***

A few of the old guard saw the calamity coming. With so many new cells in circulation, and with no means of positively identifying them, they knew there would be mixups. So they tried to head off the problem. Some learned to tell cells apart by the shapes of the chromosomes, others by how antibodies reacted to particular cultures. The methods were crude, but they worked well enough to show the scientists they had been right to worry.

One of the first cultures they checked was a human line that one day, for no apparent reason, lost its susceptibility to polio. Their identification tests suggested the cells were no longer human but had somehow been replaced by mouse cells. Then the same thing happened to a monkey cell line. Soon they found human cells growing in what should have been pig, duck, and mouse cultures. And there were mouse cells growing in rabbit cultures and rabbit cells where monkey cells should have been.

Most cell culturists were still celebrating the renaissance. But by the late 1950s many samples of the useful cell lines -- both the old, established animal cultures and some of the new human ones -- had lost their identities. An investigator who needed a normal monkey cell for his experiments could no longer be sure he wasn't working on a tumor cell from a human larynx. How could he interpret results when he couldn't even say what he had been experimenting on?

The reasons for the mess were obvious to the old guard. In a word these new people were sloppy. They were probably mislabeling cultures. Hell, they were probably using the same pipettes to feed different cultures, inadvertently picking up a few cells out of one dish and dropping them into another. If the first cells were stronger, they would grow over the second culture and replace it. Such faux pas slipped by unnoticed in many hectic labs where accurate record keeping had become a lost art. In addition, cell swapping was rampant. And when one researcher traded materials with another, he also traded mistakes.

Well, things simply couldn't go on this way, declared the veterans who had uncovered the confusion. They decided to rescue the field by setting up a central cell bank, a Fort Knox for cell cultures. Not just any cell cultures, you understand, only those with clearly defined characteristics and carefully documented histories. No shadowy pasts allowed, no vagabond cells that had wandered from one nameless lab to another. With the help of the National Cancer Institute, which was also beginning to worry about the quality of cell cultures, the group announced in 1962 that the nation's "reference cells" would be housed at the American Type Culture Collection, a private supplier of biological materials in Washington, D.C. Workers there would maintain these purebred cultures and distribute them to any researcher who wanted the very best. Over the next four years, the cell bank filled its refrigerators and incubators with more than two dozen high-quality cultures. It looked as if the science of tissue culture had backed away from the brink of chaos.

Yet as careful as they were, the founders of the bank were for years haunted by doubts. Most of their screening tests determined only what kind of animal a cell line came from. They could tell mouse from human, but they couldn't easily tell most human cells apart. What they needed were markers, chemical or structural fingerprints that were readily recognized and unique to each cell line within a species.

In 1966 a Seattle geneticist named Stanley Cartier stood up at a scientific meeting in Bedford, Pennsylvania, and offered the tissue culturists just what they needed. In appreciation, the tissue culturists practically ran him out of town.

***

As part of his study of human genes, Cartier had been looking for long-lasting cell lines that produced certain isoenzymes, enzymes that are present in every human cell but may vary in style from person to person. One of the isoenzymes of interest to Cartier was C6PD, the glucose metabolizing enzyme that comes in two styles, type A and B. Another was PCM, available in types 1, 2, and 1-2, a combination of styles analogous to the blood type AB.

Cartier first analyzed the PCM in a few of the established human cell lines and was surprised to find the same form of the enzyme in each: type 1. He tested a few more and then a few more. He stopped at eighteen. Now it is true that in an average population many people would have type 1 in their cells, but the statistics require that nearly a third of them should not. That was what bothered Cartier. When he tested the cultures for the C6PD enzyme, again they were all the same: type A. That meant that every cell line he checked had come from a black person. Most of the established cell lines, however, had reportedly been cultured from Caucasian patients. Only HeLa was known to have come from a black.

Because HeLa was the earliest successful cell line, used in virtually every lab before the rest of these eighteen cultures came along, Cartier drew what he thought was an obvious conclusion.

Oddly enough he didn't think his serendipitous finding was all that important. After all, it offered no new scientific principle or insight. The main point to him was that these variants of different enzymes could be used for telling human cells apart; they were practical tools for sorting out existing mix-ups and preventing future ones. In fact the title of the paper he submitted to the Bedford conference was "Genetic Markers as Tracers in Cell Culture." But then Stan Cartier was a geneticist, not a tissue culturist.

"My Cod, they're going to tear you limb from limb," said a friend when Cartier arrived at the meeting to deliver his report. "I can't believe what you're saying."

Cartier said it nonetheless. He stood up in the convention hall of the Bedford Springs Hotel and said that his tools for distinguishing human cell cultures demonstrated that most of them weren't different at all. He said that the eighteen cell lines he tested, samples of which were now in the vaults of the nation's new cell bank, were really just the ever-popular HeLa cells. He added, almost incidentally, that researchers who had experimented on these cervical cancer cells believing they were liver, or blood, or bone marrow, or anything else had better reconsider their findings. "The work is open to serious question," said Cartier, "and in my opinion would be best discarded."

The tissue culturists were not pleased to hear this, particularly from a geneticist, particularly since they had spent the last ten or fifteen years studying samples of these cells, thinking they were looking at many distinct forms of cancer and at normal tissue from lots of different organs. Even the founders of the cell bank who had suspected trouble found it hard to believe. And to the researchers who had actually created the cultures on Cartier's list of spoiled goods, who had toiled for years and suffered repeated disappointments before they finally got those cultures to take root, his findings were impossible. They began hurling skeptical questions.

Just how did he know the cells hadn't been taken over by HeLa in his own laboratory?

They were all analyzed as soon as he received them, Gartler answered.

But hadn't some been sent to him frozen?

We can analyze frozen samples, said Gartler.

Well, isn't it possible for a cell to change its enzyme type -- from G6PD type B to A, for instance -- and still remain the same in all other ways?

Gartler sighed. These people were obviously not geneticists. He explained that there are indeed some characteristics in a cell that may readily change. In a developing embryo, for example, a cell that had been relatively nondescript may, in the process of "differentiating," turn on various genes that manufacture different kinds of proteins. But there is only one gene in each cell that controls the production of the G6PD enzyme; that gene specifies either type A or type B, and it is always turned on.

What about random mutations?

Gartler estimated that the chances of a mutation reshaping the gene for type B into a gene for type A would be less than one in a billion. Even if so unlikely a mutation took place in a single cell, why would that single cell overtake the rest of a culture, Gartler asked. And why should such unlikely occurrences have happened in all eighteen cell lines?

Leonard Hayflick stood up. A highly respected cell biologist and an officer of the Tissue Culture Association, which was sponsoring the conference, Hayflick was also the originator of a cell line known as WISH. WISH was one of the cultures that Gartler had discredited. It was a culture grown from a scrap of amniotic sac. In fact, Hayflick told the attentive audience, WISH had been taken from the amniotic sac in which his daughter Susan came into the world. WISH stood for Wistar Institute, where Hayflick was working, and for Susan Hayflick. Since Hayflick and his wife were both Caucasian, the claim that WISH showed a genetic trait found only in blacks was a tad awkward.

In perfect deadpan, Hayflick announced, "I have just telephoned my wife, who assured me that my worst fears are unfounded."

The crowd thought that was hilarious, and the tension eased. But Gartler wasn't so sure that Hayflick was genuinely jovial. And when the laughter died down, the eminent cell biologist dismissed the geneticist's conclusions, saying simply they were very difficult for him to accept.

Then rose Harvard University's Robert Chang, another luminary of cell biology and a trustee of the Tissue Culture Association. Chang was the creator of one of the most popular of human cell lines. The Chang liver culture was used extensively in studies of liver function.

Whatever culture Gartler claims to have analyzed, said Chang, it wasn't a culture that came from Chang. "I have never sent him any cell line, and I don't remember ever having corresponded with him."

Gartler explained that one of his samples of Chang liver had come from a co-worker at the University of Washington in Seattle. The other came directly from the cell bank at the American Type Culture Collection. In fact, six of the eighteen cultures he examined had come straight from that storehouse of only the best and most carefully screened cultures. The important point, said Gartler, is that while there may well be some genuine samples of these cultures at certain laboratories, there are others in active use that are impostors. Unless experimenters can tell the bona fide from the bogus, he said, much of the research done on these cultures is in doubt.

It looked to Gartler as if Chang was contemplating murder -- or suicide -- but all he did was sit down.

More skeptical questions, more icy speeches. The session finally ended at noon, Gartler escaped from the room, and the tissue culturists changed their tactics. Now it was a war of isolation. They ostracized him for his wild and insolent claims. Through most of lunch he sat alone. One of the few who dared to join him was a young researcher from California. As a technical advisor to the cell bank, the researcher was as disturbed as the others about Cartier's findings. At the same time, though, he admired Cartier for sticking his neck out, and he told him so. The researcher's name was Walter Nelson-Rees.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:23 am

4: Out of Thin Air

It wasn't until two years later that Gartler's incredible conclusions were confirmed. By 1968 two independent research teams had applied his methods to all the human cultures deposited in the cell bank at the American Type Culture Collection. Out of thirty-four cell lines, they found twenty-four to be HeLa.

How could it have happened?

The most likely explanation was the same combination of sloppiness and opportunity that in the late 1950s had shuffled mouse cells with human cells and duck cells with monkey cells. HeLa had a number of advantages that helped it pull the trick off on a disastrously large scale. Since it was the first useful human cell line, it was the most ubiquitous: wherever technicians were careless, there were always a few HeLa cells nearby to take advantage. Because it was also one of the most vigorous cultures known, it could easily take over weaker cultures if given half a chance.

HeLa was so tenacious, in fact, that it probably didn't need to wait for a lab worker to use the same pipette on different cultures. A startling series of experiments reported in 1961 by Lewis Coriell, one of the old guard who had helped start the reference cell bank, had shown HeLa could literally appear out of thin air. Coriell, working at the Institute for Medical Research in Camden, New Jersey, found that merely pulling a stopper from a test tube or dispensing liquid from a dropper could launch tiny airborne droplets containing a few HeLa cells. When the drops landed on open petri dishes holding live cultures, the HeLa cells began growing so feverishly that in three weeks they overwhelmed the original cultures.

To some it had seemed an unbelievable observation. But now it looked as though much the same thing must have happened to all the human cultures that appeared in the 1950s soon after HeLa. Those cells had probably been as weak and hard to cultivate as the ones that had been tried in the pioneering days; they were easy victims for HeLa. A few of the more cynical scientists suspected there had never been anything but HeLa in those new cultures. The cells from the original tissue samples had probably died immediately, leaving a culture dish of nutrients ripe and ready for the next HeLa cell that happened by.

Either way, for Coriell and some of the other experts, HeLa's surreptitious spread explained a few puzzling observations that cancer researchers had made in recent years. One such enigma was "spontaneous transformation," a mysterious process by which benign cells suddenly turned malignant. There it was right in the dish, the very nut of the cancer problem: healthy cells, going along in a calm and orderly way, abruptly burst into unbridled growth. Not only did they grow faster, they were no longer bound by the normal cells' lifetime limit of fifty to sixty divisions. The transformed cells ignored their biological clocks and continued doubling without end.

The weird thing about spontaneous transformation was that until the late 1950s and early 1960s, it was never known to occur in human cells. Cultures of rat, mouse, hamster, and other animal cells had been spontaneously transforming for years, but never a human cell culture. Then suddenly it was happening all the time. Not only that, these spontaneously transformed cells grew rings around many cells taken directly from patients' tumors. Nobody could explain why normal cells that turned malignant in the lab should be more aggressive than cells that had become malignant while in the body, but for the moment scientists were delighted to have all these tenacious new cultures.

Much later it became clear that these transformations were not spontaneous at all, but had been triggered by outside agents. In the case of the nonhuman cells, chemicals in the nutrient medium, oxygen in the air over the cultures, even fluorescent lighting in the laboratory were eventually found to inflict genetic harm that can turn normal cells cancerous. As for the human cells, most "transformations" appeared to be nothing more than takeovers of the cultures by the feisty HeLa cells.

Spontaneous transformation was not the only myth HeLa created about the nature of cancer. Researchers had observed that cancer cells shared many fundamental characteristics, and there had begun to emerge a unifying theory: all cancer cells grew relatively quickly and had the same basic nutritional requirements; they seeded new tumors when inoculated into the cheeks of hamsters; many had abnormally shaped chromosomes; and most carried the same surface antigens, proteins on the outside of the cell that stimulate the body's immune system. Like winning lemons in a casino full of rigged slot machines, these traits kept coming up one after another in dozens of cell lines the scientists thought had come from dozens of cancer patients. The truth was they had been studying one line of cells masquerading as all the others, and the common traits they saw were those of a single tumor, the one that killed Henrietta Lacks.

"They described a lot of things they thought were being produced by intestine and kidney and other cells," recalled Coriell years later. "There was a lot of data in the literature that was just wrong, just a lot of wasted time because they were all working with HeLa."

Cyril Stulberg of the Child Research Center in Detroit, like Coriell one of the deans of cell culture, came as close as anyone to assessing HeLa's effect on this early period of cell biology and cancer work. He made the remark in a letter to Nelson-Rees many years after Cartier's HeLa revelations. "I didn't realize then what the succeeding years would bring," Stulberg wrote. "Naturally, at the time, I was very defensive because I saw 15 years work go down the drain."

But Stulberg, Coriell, and the other veterans faced up to the calamity and began to clear away the wreckage. As the architects of the fledgling central cell bank, they were reluctant to simply throw out the twenty-four HeLa-contaminated cultures. HeLa, after all was a sturdy line and these individual strains had various quirks and characteristics that made them particularly useful to certain fields of research. The solution, they decided, was an addendum to the cell bank's catalogue in 1968 warning that the twenty-four lines were actually HeLa and should be used as such. Moreover, they required even more detailed descriptions of new lines deposited in the bank and recommended not only Cartier's enzyme tests but any other promising techniques of identification as well.

This time, they hoped, they would put HeLa contamination and all the other chaos behind them. And, indeed, as the 1960s came to a close, it appeared that the cell biologists had recovered from Stan Cartier's bombshell. Cartier himself went back to studying genetics, having done quite enough for cell culture.

***

As for what made HeLa so damned stubborn in the first place, the scientists could only speculate. In 1970 two researchers dug out the original biopsy slides of Henrietta Lacks's tumor and uncovered one possible answer. The pathologist at Johns Hopkins in 1951 had described her illness as the most common type of cervical cancer, involving "epidermoid" cells, which form a skinlike covering over the cervix. He had made a mistake. It was clear to the researchers who reviewed the slides twenty years later that the tumor was made up of "immature glandular" cells. Such tumors were thought to be far more aggressive in the body. That would explain the speed with which Henrietta Lacks's cancer killed her. Maybe HeLa's widespread contamination of other cell lines demonstrated that immature tumors are aggressive in culture as well.

Another theory held that though HeLa started out as a pretty scrappy scavenger of petri dishes, years of breeding in laboratories all over the world had made it even scrappier. As investigators subjected HeLa cultures to different stimuli -- X rays, for example, and viruses -- single cells must have changed, some becoming still stronger, others turning delicate. The fragile variants would have died off, leaving only those most fit for life in the lab. A corollary of this theory was that new strains of HeLa would have emerged with personalities the original HeLa cell never had. Though it wasn't yet generally known, there were indeed new subfamilies of HeLa cells making the rounds.

One of these novel strains was quietly touring Europe in the late 1960s and early 1970s. It had started as a standard HeLa culture, probably sent by an American scientist to friends abroad, who then passed it around. At one of the labs it visited, this HeLa culture contaminated a culture of monkey cells, killing off the monkey cells but inheriting a colony of viruses that had infected them. It first surfaced in 1973 when a group of West German scientists reported the surprising presence of monkey viruses in a human cell line. But by then, it had traveled extensively, no doubt hopping in and out of culture dishes under a variety of aliases. When researchers tried to reconstruct the wanderings of this HeLa variant many years later, all they could say with certainty was that it must have slipped into the Soviet Union at some earlier point -- because it was this strain of HeLa cells growing in six separate cultures that the Russians handed back to the Americans in 1972.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:23 am

5: In the Purple Palace

It wouldn't be right to say that the officials of the National Cancer Institute were surprised when Nelson-Rees delivered the bad news in the spring of 1973. Given the sorry state of Russian research, it came as no shock; that was the scientific point of view. Being bureaucrats, however, the institute people saw this as a political mess fraught with ironies that would delight the press and other antagonists. It was bad enough that the earliest fruit of Nixon's scientific detente was the discovery that the Russian cultures were good for nothing. But for the cultures to have been spoiled by the cells of some posthumously hyperactive woman from Baltimore, cells that had probably slipped into the Soviet Union as the result, however indirect, of some American researcher's largesse many years earlier -- that was too much. No, they were not surprised. They were mortified. If true, the news of Henrietta's Russian appearance would be a diplomatic nightmare.

Well, then, maybe it wasn't true.

They urged Nelson-Rees to check his results. They cautioned him not to say a word to anyone. They suggested this could jeopardize the future of the entire bio-medical exchange with the Russians. And reminding him that he had broken the rules by sending the cells to Detroit without permission, they extracted a written apology from him. But along with the apology Nelson-Rees sent the evidence, summaries of the many tests and retests. By early fall, the bureaucrats had no choice but to begin quietly discussing how best to break the news to the Russians. The Soviet delegation was due to visit the institute's headquarters in Bethesda, Maryland, in a few months, just as the American entourage had visited Moscow a year earlier. One of the main events scheduled was a review of what the Americans had learned about the Russian cell cultures.

In October Nelson-Rees received a phone call from John Moloney, assistant director for viral oncology at the institute and the leader of the American delegation to Russia. Never before had Nelson-Rees received a phone call from Moloney. Moloney was just one step down from Frank Rauscher, the director of the institute. And Rauscher, unlike the directors of other U.S. health institutes, was appointed by the president himself. It was a little like a sixth-century English footsoldier getting a call from Sir Lancelot.

Moloney said he would appreciate it if Nelson-Rees would come to Bethesda to make a presentation to both institute officials and the visiting Russians. The topic would be the "possible HeLa origin" of the Soviet cells.

"Of course, we must keep this information from becoming offensive to our Russian collaborators," he added. "Especially to Doctor Zhdanov." Victor Zhdanov was Moloney's counterpart, the senior scientist on the Russian side who had organized the presentation of the cell cultures.

Nelson-Rees felt honored by the request. But as the excitement of the phone call wore off, as he looked over his notes of the conversation, he also began to feel vexed. "Keep this from offending the Russians." It occurred to him that no one was genuinely interested in his work. As far as Nelson-Rees could tell, Moloney's deputies had for the past few months concerned themselves only with the awkward ironies and procedural infringements that it had stirred up. Now even Moloney's personal invitation seemed to him largely a warning that we must not insult our comrades in science.

Come on now, thought Nelson-Rees, this discovery resulted from a dandy piece of detective work. Not only that, it meant that HeLa contamination was still a problem -- in fact it appeared to be a worldwide problem. What about a little acknowledgment? What about a little fanfare? What about my publishing it in the scientific press if you folks aren't interested? Or is the protection of Russian egos more important than spreading the word that Henrietta Lacks is still at large?

But in a letter to Moloney a few weeks later Nelson- Rees did not express himself quite so crassly. He outlined the material to be covered in his talk and added simply, "I would like to submit these results for publication as soon as you feel that the necessary amenities have been extended to all parties concerned."

***

On 12 November 1973, Nelson-Rees found himself in conference room number 10 at the headquarters of the National Cancer Institute in Bethesda, ten miles north of Washington, D.C. The classiest of the institute's meeting places, it was nicknamed "the Purple Palace" for the mauve carpeting and upholstery that adorned its outer lobby. A forty-foot table of heavy oak dominated the room. The table was oval and had a band of black leather along its edge upon which conferees could rest their elbows. There were microphones to amplify voices and earphones to offer simultaneous translations. The audio technicians and translators sat in a soundproof booth behind a darkened window set in one of the solid oak walls. The ceiling was made of wooden slats arranged in a waffle pattern. The carpet was thick and bluish gray. The chairs were black leather recliners with wheels.

On 12 November, for the meeting of the U.S.- U.S.S.R. Joint Subcommittee on Research in Oncologic Disease, the table was bedecked with miniature flags. Thumbelina versions of Old Glory and the Hammer and Sickle stood side by side, sticking up out of wooden hockey pucks in front of each namecard and water pitcher. On one side of the table sat the Americans, John Moloney and Nixon's appointed anticancer crusader Frank Rauscher at the center. On the other sat the Russians, arranged around Victor Zhdanov. Walter Nelson- Rees sat behind the American side of the table in a kind of raised gallery, the bleacher seats. About an hour into the meeting, Moloney looked over his shoulder and motioned for Nelson-Rees to come forward.

"We have examined the chromosomes and G6PD mobility patterns of six cell lines received in our laboratory in January 1973 from the Soviet Union via Bethesda," Nelson-Rees began. He was nervous. Like a kid putting a lit match to a pack of fire crackers, he was both looking forward to and dreading the explosions he planned to set off around him.

"Quinacrine-fluorescence indicated the absence from all cells of a Y chromosome." (Translation: We couldn't find a single Y chromosome in any of these cells. Although there are other explanations, this suggests all the cultures came from females.)

No reaction. He could see Victor Zhdanov staring at him: crew cut, mustache, and spectacles, but no expression on his face.

"G6PD mobility for all lines was of type A variant, characteristic of HeLa cells." (All six cultures contain the rare form of the G6PD enzyme, a form found only in the black population. Isn't that odd? Every cell seems to have come from a female who was black.)

Maybe someone coughed. Several of the American delegates may have nodded, their faces blank.

Nelson-Rees placed an enlarged black-and-white micrograph of chromosomes on the easel at the head of the table. Chromosomes labeled as coming from the Russian cells were arranged in vertical columns next to chromosomes from known HeLa cells. Barber pole patterns were clearly visible on all the chromosomes. "A trypsin-Giemsa technique for chromosome banding revealed marker chromosomes common to all cell lines, as well as to our culture of HeLa cells and to HeLa cells reported in literature." (As you can readily see, the fingerprints of the Russian chromosomes match those of the HeLa cells. The cells are identical.)

Dead silence.

It was as if he had been up there telling a long, complicated joke and had neglected to deliver the punch line. "Oh-huh, and then what happened?" several faces seemed to say.

There were others who refused to look at him. They riveted their eyes on the table before them, as if they were ashamed of something, as if they were hoping he would just finish the talk and spare them anymore of this crude, unspeakable subject.

There were also one or two faces fighting back the urge to smile at the whole humorous mess these Russian cells had been, but those faces were turned away for fear of breaching the solemnity of the occasion.

Nelson-Rees couldn't believe it. Maybe they simply failed to understand the data. Many of the Russian delegates were not well versed in the techniques of tissue culture. Yes, perhaps the significance just escaped them. Or maybe the Americans were still peeved that he had overstepped his bounds in checking out these cells and had risked unleashing a hoof-and-mouth plague on Detroit: Then again, maybe the whole lot of them had decided the most diplomatic way of handling his discovery was to ignore it.

Nelson-Rees had been girding himself for an explosion, and there wasn't even a burp. It was embarrassing. He felt suddenly silly up there at the easel in front of all these zombie-eyed scientists. Moloney thanked him for the presentation, and the meeting of the U.S.-U.S.S.R. Joint Subcommittee on Oncologic Disease was adjourned for lunch.

***

It was Victor Zhdanov who finally asked, "What does it mean?"

Zhdanov, Ne1son-Rees, Moloney, and a few others had gathered after lunch in Moloney's office. It was quickly apparent to Nelson-Rees that the earlier session had been ceremonial. Now the Russians would have a more private and informal setting in which to discuss these curious findings. Under other circumstances Ne1- son-Rees might have been loudly indignant for being set up as part of the flag show in the Purple Palace. But he was so anxious to describe his work to anyone honestly interested that when Moloney asked him to repeat the highlights of his talk, he did so cheerfully. Then, when he had finished, Zhdanov asked his question.

"First of all/' said Ne1son-Rees, "it means you are not in control of the situation. You do not know what cells these are. You seem to have no control over where they come from or where they are going. But more importantly these cells are one and the same, and the viruses are one and the same, and it just isn't right to be calling them six separate isolates of human cells, each carrying a virus with potential human cancer value."

This time there was no stony stare. Zhdanov was visibly upset, though it is hard to say precisely what was racing through his head. He may well have been picturing the long list of Soviet research reports that were based on examinations of these cultures. In effect, Nelson-Rees was saying that most of the Russians' work on viral cancer in the last few years was questionable at best. Whatever he was thinking, Zhdanov quickly sought to take some advantage out of the catastrophe. "If you know so much about cells and can do so good," said the Russian, "why don't you come to Moscow? Show our people. Talk about what techniques you have."

"I'd be delighted," shot back Nelson-Rees, "if you promise I can get back out of the country."

The Russian laughed loudly.

Nelson-Rees joined him.

Suddenly it was one big happy meeting. Nelson-Rees jostled and joked further with Zhdanov. With some of the other Soviet delegates, he discussed opera and ballet, including the work of Rudolf Nureyev, a Soviet defector. If a few of the American bureaucrats were still sore about Nelson-Rees's independence in the handling of the Russian cultures -- and miffed anew at his accepting this invitation to Moscow without consultation -- they didn't let on. Finally, thought Nelson-Rees, the world is regaining its sanity.

***

The Journal of Virology soon proved him wrong. Now that the "necessary amenities" had been extended to "all parties involved," Nelson- Rees was set to publish his work. The friendly conclusion of the Bethesda conference had made him optimistic; the joint committee had gone so far as to formally agree it was "extremely important" that the findings be published. He prepared a manuscript and submitted it to the journal, which had just printed Wade Parks's conclusion that the Russian cells contained only monkey viruses. Certainly the journal would want its readers to know that the cells themselves were not what the Russians had claimed either.

The people at the journal didn't see it that way. "First of all it seems to be a gratuitous attack on the Russians," wrote one of the two technical judges of Nelson- Rees's report. "Secondly (and more importantly) it really is a footnote to history and not a study with new scientific implications .... I would hope ... someday it would appear as a footnote." The other judge also suggested that the finding be presented only as a short blurb, and he preferred that it be presented in somebody else's journal.

Nelson-Rees was stunned and furious. Were these people -- these people who are working scientists devoted to communication among colleagues -- were they as worried as the bureaucrats about the politics of unpopular findings? It was the same incredible blindness. They couldn't see what was so obvious to him: the scientific community must be warned. The word must be spread that Gartler's list seven years ago was not the end of the story and that the cells of Henrietta Lacks are still on the loose!

When Parks heard the news, he wrote to the journal's editor, gently recommending that he publish some form of Nelson-Rees's paper. The editor sent a note to Nelson-Rees saying that having thought about it further he "would not be averse" to publishing a much shorter version of the report.

He would not be averse.

Nelson-Rees was not the sort of fellow to tell anyone to roll up a manuscript and stuff it, but he came close to making the suggestion to the editor of The Journal of Virology. Seething, he withdrew the paper and sent it to The Journal of the National Cancer Institute. The judges there were not as hostile, though several had trouble grasping the importance of the paper. It was finally accepted in June of 1974 and appeared in print three months later, almost two years after the Russian cells arrived in the U. S. and a year after the meeting at the Purple Palace.

The whole experience left Nelson-Rees sour. He had been wrong to think that anyone wanted to hear this bad news, except, ironically, the Russians. The funny thing is that the entire frustrating misadventure tempted him to jump in with both feet. It was almost enough to start him off on a personal crusade. Almost.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:24 am

6: Keeper of the Cells

Growing up in Germany in the 1940s, Walter Rees was an outsider. He had been born in Havana and lived there until his parents separated when he was nine years old. Walter's mother then took him and his older brother to Karlsruhe, a town on the Rhine River. Living in Germany was the only way they could claim a small inheritance, left to his father, that would finance the boys' schooling. Walter's father, a German who sold farm equipment to sugarcane growers and later owned the Pepsi-Cola bottling plant in Havana, decided to remain in Cuba.

Being the sons of a German man, Walter and his brother were considered by the Third Reich to be German themselves. The term was Auslandsdeutscher, an outland German. It was an appropriate description for a young German boy who spoke only Spanish and English.

Nine months after Walter and his brother began classes at a strict, religious boarding school in southwestern Germany, Hitler invaded Poland. The Rees boys, like all the other children, joined the Tungvolk, the cub scouts of the Hitler Youth. Young Walter didn't enjoy the marching and the patriotic singing the way many of his classmates did. When he heard about plans for a military camp out in the country that summer, he announced that he wasn't going. An older boy, one of the group leaders who held the title of Fuhrer, beat him severely for presuming he had any choice in the matter.

Walter was a good student. He learned to be disciplined and neat. He could do anything they asked of him; he just didn't feel a part of it. And he was happiest when he escaped. Whenever he could, Walter ran off to a section of the Black Forest that surrounded his school. There in the woods he and a friend built tree houses and pretended they were exploring the dark continent. Sometimes Walter would tell stories out of adventure books he had read or describe the real-life dreamland of Cuba.

No one spat at Walter as they spat at the little Jewish boys who were forced to wear Stars of David, and he was thankful for that. But as the war intensified and the Americans entered to fight Germany, he felt a prejudice against him and his family just the same, a sense that everyone knew they didn't belong. His mother, having been born in Havana during the U.S. intervention that followed the Spanish-American war, was considered to be an American citizen. Once when she was travelling to Messkirch, an official swept through the train, demanding to see identification papers. That was not unusual in wartime Germany. When he arrived at her compartment, however, the man smiled and said, "That's all right, you needn't show us anything. We knew you were riding with us, Frau Rees." They had been watching her.

Walter's mother taught him to be polite, helped him discover art and music, and encouraged him to be very good at whatever he did. She never said it quite this way, but what Walter heard was: be perfect. Show them that even if they won't accept you, you can be just as good as they are. Years later when Walter was in college, his mother tried to kill herself with sleeping pills. He thought that somehow he had disappointed her. "That led me to want to excel at many things," he told a friend. "I wanted to do everything at least as well as the best around."

Walter was sixteen in 1945 when the American forces arrived to occupy Germany, though to him it seemed more that they were liberating the country. After years of rationing, suddenly there were chocolates and cigarettes and fresh fruit. What's more, there was no school. Walter got a job with the 485th U.S. Army Medical Collecting Company, a kind of mini-hospital that had set up near his home in Karlsruhe. He translated orders, typed letters, and washed and sterilized medical equipment. Although he had never been particularly interested in science, he decided then to become a doctor. After the war Walter returned to Havana, finished high school, and left for Emory University in Atlanta.

He dropped his medical ambitions after fainting in a dentist's office while a friend was having a tooth pulled -- the sight of the blood had been too much for him -- but he was already hooked on the study of plants, insects, and cells. He would sit at a microscope mesmerized as the cells of various animals doubled and redoubled before his eyes. Grasshopper cells put on the best show because they had the biggest and most visible chromosomes. That was what he fancied most, the chromosomes, those mysterious strands of genes that threaded themselves into intricate knots, emerged in orderly pairs of original and perfect duplicate, fell into formation across the center of the cell, and then marched to opposite ends, cueing the cell to split in two. In the spring of 1951, a few months after Henrietta Lacks's cervical tumor cells were put into culture, Walter Rees graduated with a bachelor's degree in biology. A year later, having earned a master's in cytology, also from Emory, he joined the U.S. Army. He had decided to remain in the United States, and the only way to quickly become a citizen was to enlist.

After three years at Dugway Proving Ground in Utah, working in the Army's biological weapons program, he received his citizenship, changed his surname to include his mother's family name, Nelson, and headed west, enrolling in a doctoral program in genetics at the University of California at Berkeley. There, under the guidance of Spencer Brown, a world renowned researcher who later became president of the International Genetics Federation, Nelson-Rees turned his idle fascination with chromosomes into serious study. Brown, known to some as Mr. Chromosome, taught Nelson-Rees the importance of finding and memorizing the chromosome patterns in certain cells, especially the oddball patterns. Look for the oddballs, said Brown, they're the clues to something interesting and useful.

Brown was an impatient man with fanatically high standards for laboratory technique. As exacting as Nelson- Rees had always been about his work, he found a new level of perfection in Brown's style, and he knew he'd never last if he didn't assume the same zealous approach. He found the challenge exhilarating, and he excelled. He became the favored member of Brown's retinue of graduate students. He actually looked forward to exams so that he could show his stuff. He began to talk of the beauty and elegance of a well-prepared slide of chromosomes. And he became known for leaving parties at ten o'clock to check a fly colony, count mealy bugs, or do some other chore in the lab, where he would end up spending most of the night. It was no hardship for him to be so devoted to his experiments. The truth was he felt there were better things to do than to dull his brain cells with alcohol and small talk.

When he had completed his doctorate in 1960, the chairman of the cell culture department at Berkeley's School of Public Health invited him to join a new lab they were setting up in Oakland to support the national cell bank, the one at the American Type Culture Collection in Washington, D.C. At first he applied his skills to the identification of animal cells by their chromosomes. In the late 1960s, when "The War on Cancer" heated up and officials at the National Cancer Institute wanted to rebuild the wreckage that Stan Gartler had left behind, they asked the Oakland lab to initiate and collect new human cell cultures to be used in the institute's most ambitious endeavor, the viral cancer program. That was where the money was. That was where the action was. And there was Nelson-Rees, quietly working his way up to the top of the new Oakland cell bank.

By the early 1970s, the institute was relying more heavily on the Oakland lab, and on its new director of cell production, Nelson-Rees, than on the American Type Culture Collection. If his people didn't have what the nation's top cancer researchers asked for, they went out and got it. Thymus one day, spleen the next, and tumor tissue from Asian and American Indian patients the week after. These institute scientists also depended on Nelson-Rees to check the identities of cultures they obtained from other labs. Those familiar with the Oakland operation knew it was Nelson-Rees's meticulous manner that made it so successful, so reliable.

"Walter's one major attribute," the institute's Jim Duff once said, "is perfection."

Wade Parks, the virologist who helped analyze the Russian cells, put it this way: "Walter's a little blunt sometimes. But he's never wrong."

There was, however, a joke about the status of projects such as the Oakland lab, projects that were funded by the institute but were not conducted by institute personnel as part of the in-house program. They were called the outhouse programs. Nelson-Rees used to repeat the joke goodnaturedly, though he did feel a distance between himself and the project leaders. There was no doubt that they knew who he was and appreciated his work, but in many ways he was still an Auslandsdeutscher.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:24 am

7: Mug Shots

Officially they called it the Cell Culture Laboratory of the University of California at Berkeley, but in fact it was located in Oakland. Not in Oakland's fashionable residential hills but in the western flatlands, the marine industrial district. This was the part of town you saw first as you drove from San Francisco across the Bay Bridge: the storage tanks, the warehouses, the big Rustoleum red containerships at the docks, and, looming over it all, the gigantic metal cranes like a herd of mechanical dinosaurs foraging among the cargo at the water's edge. You knew you were in Oakland's harbor district when the roadside billboards stopped trying to sell you Johnny Walker Red and started pushing Tasco industrial valves.

The cell culture lab was housed in a former mess hall within a three-square-mile supply compound run by the U.S. Navy. There were no signs of Berkeley's rolling green lawns, the quads where students gather to picnic and play music, or the majestic marble buildings of higher learning. Here the office buildings were gray with dirty windows. Huge anchors and propellers were strewn around aluminum warehouses. In the alleyways stood dingy yellow forklifts. Scores of cargo containers, each the size of a Greyhound bus, were stacked high behind chainlink fences topped with barbed wire. Down the road was an outfit that hauled these "piggybacked" containers by rail and truck. It was called the Southern Pacific Golden Pig Service.

And yet inside the white building that held the cell culture lab there was no hint of the surrounding clutter. There was order and, through a certain set of double doors, there was cleanliness beyond imagination.

You couldn't go through the doors -- they were locked -- but you could peer through their windows into a bright, gleaming hallway. To get to that hallway you had to step into what looked like a large closet next to the double doors. Inside, a wooden bench ran from one side of the closet to the other, blocking your way. Where you entered, the floor looked like most institutional floors; its pattern of charcoal gray swirl was dulled by a hazy buildup. But just past the bench, it was a polished gem, the proverbial "floor you can eat off." On the wall was a large black-and-white sign:

Observe Change Rule
Never Come into Laboratory with Street Shoes Alone


The sign hung next to a series of shelves with forty-eight cubby holes, each labeled with the name of a lab worker, each holding a pair of shoes. Staffers had to change out of their street shoes and into a pair they wore only in the lab. For visitors there were white nylon booties in small, medium, and large.

The trick was to sit on the bench, put on a clean shoe (or slip a bootie over a dirty one) and swing it over to the clean side; then repeat the process for the other foot. Lab workers found the maneuver quite natural. But visitors often had trouble, accidentally bringing a clean bootie down onto the dirty side, in which case a new bootie was called for; or landing a dirty shoe on the clean side, which required a quick swabbing of the deck; or nearly falling off the bench in an attempt to avoid doing either.

The door in the clean section of the closet led into the gleaming corridor. Several individual lab stations extended along the hallway's left side. At the near end was the assay area, a large open space where newly arrived cells were logged in, examined without opening the flasks or dishes that held them, and stored until they could be analyzed more thoroughly and in the security of one of the "clean rooms" a little farther down the hall.

Each clean room was really a room within a room. The anteroom, about the size of three telephone booths, was where two technicians in surgical masks, caps, gowns, and gloves prepared for the task at hand, readying equipment, squirting alcohol over bottles of nutrient solution to kill bacteria, removing sterile wrapping on the glassware they planned to use. They then entered the pristine inner sanctum, closed the sliding door, and set to work: perhaps feeding a cell culture, transplanting portions of it into other flasks, or replacing the liquid medium with a fresh supply -- changing its diapers, as the crew described it. The inner room had its own source of filtered air that kept the pressure there higher than in the rest of the lab, preventing airborne undesirables from drifting inside. When the door was opened, the air always rushed out of the room, never in. The technicians worked in pairs because it eased the load of monotonous chores. Open the flask, withdraw 10 ccs of medium, close the flask, label a new flask, open it, dispense 10 ccs of medium .... Working in pairs they could also keep an eye on each other. They were not to talk unless absolutely necessary; in spite of masks, talking increased the chance of spitting out microorganisms that might endanger the cell cultures. To eliminate the risk of cross-contamination, they worked on one cell line at a time, re-sterilizing the whole show before another line was brought in. It was an expensive, time-consuming procedure. They stayed in there for two to three hours at a time. But then obsessive care was what made Oakland so special.

When work on a particular culture was finished, any materials to be discarded were placed in pans full of disinfectant and taken to the far end of the hallway to the autoclave, a little chamber of hell that subjected its contents to lethal temperatures and pressures. All glassware was sent to be washed and rinsed in scalding, deionized water. Every working surface in the clean room was then wiped down with alcohol. Just in case some scrap of life remained at large in the room, and to kill off anything that might creep in later on, one of the technicians flipped a switch on the way out, bathing both compartments in ultraviolet light. On the door to each clean room was a sign that said in red letters:

KEEP CLOSED
Admittance to authorized personnel only.
Visitors and personnel not assigned to this area
contact W. Nelson-Rees.


On the other side of the corridor were the cramped quarters where Nelson-Rees and his co-workers mulled over the results of the lab work. Bob Flandermeyer had the office next to Nelson-Rees. He did most of his mulling while hunched over a table, pawing through a pile of photographs like a man searching for the right piece in a jigsaw puzzle.

***

Chromosomes, chromosomes, you could always find Bob Flandermeyer at that table sorting through hundreds of black-and-white enlargements of chromosomes. Long, straight chromosomes; short, stubby chromosomes; chromosomes that looked like bow ties, black ants, and licorice twists; and all wearing stripes, the irregular barber pole stripes known as banding patterns. Slowly, deliberately, ploddingly, Bob Flandermeyer would cast his big soft eyes over each photograph, studying the shapes and patterns.

Flandermeyer was a bear of a man with blond hair and a wide, friendly face. He spoke softly and a little ponderously. His laugh, loud and sharp, was startling punctuation to his sleepy speech. In early 1973, he left his job as a technician in a neighboring Navy laboratory. He had been working on a study of how bacterial diseases spread among troops, which had little to do with cell biology. Nevertheless, Nelson-Rees hired him to help apply the technique of chromosome banding to the identification of cells. Two other lab assistants before Flandermeyer had tried and failed to make it a practical system. The work was too tedious for his predecessors, but Flandermeyer mastered it.

The first part of the technique was nothing more than what karyologists, chromosome analysts, had been doing for years. The idea is to catch the cells in a stage of their growth cycle called metaphase, when the chromosomes line up across the center of each cell in orderly rows. You do that with colchicine, a drug derived from crocuses. Colchicine prevents the formation of tiny fibers that normally attach to the chromosomes and drag them to opposite ends as a cell prepares to split. It halts the chromosomes in their lineup like dancers in midstep. Then you add a solution to make the cells swell, which spreads the chromosomes apart so they don't overlap, and then an alcohol "fixative" to kill the cells. The next step is to attach a drop full of cells onto a microscope slide and flatten it into a layer thin enough that everything will be in focus for the camera. One popular method of flattening was the five-foot drop: holding the dropper at eyelevel, you aim at a group of slides arranged around your feet. From that height, a drop spreads itself into a broad, thin film, and if you're lucky the film ends up covering a slide instead of your shoes or the linoleum. For those with more flamboyant tastes, there was the crepe suzette technique: a match is put to the slide, igniting the alcohol fixative, which burns away in a few seconds, dramatically reducing the volume of liquid in the drop. Once the sample is properly flattened, you mix in a little Giemsa stain and place the slide under a microscope. The chromosomes show up in silhouette, dark blobs without detail. Based on their shapes, a good karyologist can tell the species of the donor: man, mouse, or whatever. By studying the shapes and counting the number of chromosomes, he can also tell if the cells are abnormal. Victims of Down's Syndrome, for instance, have one more than the usual human complement of forty-six chromosomes.

That was conventional karyology.

In the new banding technique, before staining the cells, you dip a slide full of them into trypsin, an enzyme that eats away parts of the protein coat over the chromosomes. Then, instead of painting the entire outer coating of the chromosome, the Giemsa stain is absorbed into it, but only in certain spots, creating discrete bands of darkness next to blank areas. The problem is that the trypsin and the stain have to be at particular concentrations and temperatures, and the slides have to be dunked into each solution for precise periods of time. Even if you figure out the proper combination, the technique doesn't work unless you catch the cells at just the right moment in the metaphase stage. For some reason, the enzyme can't eat through the chromosome's coating otherwise.

Flandermeyer studied several days with a group of researchers who were using the technique at the University of Indiana. He returned to Oakland and began experimenting. Time and again he would prepare a bunch of slides, dip them in trypsin while counting off the seconds, then dunk them in the stain, only to find the familiar dark blobs where he had hoped to see barber poles. That meant he had exposed the cells to the enzyme for too long, allowing the stain to seep in everywhere. Or had he got the temperature wrong? Other times, when he didn't keep them in long enough or failed to catch the cells in metaphase, no readable patterns appeared.

Finally he hit on a reliable recipe for producing good, clear bands. He stained stacks and stacks of slides, then sat down at the microscope and scanned one after-another, searching for the most photogenic cells. He photographed hundreds of them, printed up enlargements, cut out each banded chromosome, and mounted them all on white cardboard, grouped according to their banding patterns. After that the real work began.

First he had to memorize the patterns of normal human chromosomes. Every healthy cell has two copies of chromosome numbers 1 through 22, one from each parent, plus either a pair of Xs in the case of females or an X and a Y for males. So there are twenty-four different types of chromosomes, each type absorbs the stain in a characteristic set of bands -- its mug shot -- and Flandermeyer had to know each one on sight. He stared at the photographs for weeks. Through brute force of will he developed an eye for all twenty-four. But because these were normal chromosomes, present in every human cell, they were really no help in telling cell lines apart -- not directly. He needed to know the normal chromosomes only so that he could recognize the misfits, the ones with funny shapes and weird banding patterns.

The misfits, the theory went, resulted from random errors in the duplicating process. Such errors occur frequently in tumor cells, where the usual mechanisms of replication and growth run amok. The lower arm of a number 5 chromosome might break off and attach itself to a severed upper arm of a number 3, for example. This unlikely goat with a lion's head, passed on to daughter cells, becomes a unique marker of that particular tumor and of any cell line derived from it. The odds were slim that an identical misfit would arise by chance in another cell line. And it was close to impossible for one cell line to randomly create a set of three or four misfits that matched those in another culture. Only cells with a common origin would have the same aberrant marker chromosomes.

For Nelson-Rees, who had always been partial to chromosomes, and who had learned the value of misfits from Mr. Chromosome himself, these markers were among the more powerful means of identifying human cell lines. When the National Cancer Institute made the Oakland lab its hot new repository for human cultures, he decided that his quality control arsenal had to include a method of recognizing markers. But conventional staining wouldn't do it. To spot a marker you have to be able to make out which pieces of normal chromosomes had combined to form it. For that level of detail, you need to see the bands. That was why Nelson-Rees needed Bob Flandermeyer. And that was why Bob Flandermeyer, having learned every normal banding pattern, began staring at photographs of the abnormal markers, forcing his mind to associate each strange patchwork pattern with the cell line it had come from. The first markers he learned to spot were the four that other researchers had found in HeLa cells.

My God, he's slow, thought Nelson-Rees as he watched Flandermeyer pick his way through the photographs of unidentified cell cultures, looking for markers he had already memorized and memorizing the ones he came upon for the first time. "Ein Stier," Nelson-Rees had once called him, "an ox."

There was considerable art to the method, though. Flandermeyer knew that a banded chromosome's appearance often depended upon the angle from which he photographed it and whether it happened to be drooping or twisted or contorted in some other way. There was a lot of visual judgment involved, and Flandermeyer was not one to jump to conclusions.

Nelson-Rees and some of the other researchers who had learned to read banded karyotypes with Flandermeyer's help would often ask him to confirm their own assessments. He was, after all, the expert. On these occasions, Flandermeyer would stare at the photographs endlessly as the rest of them paced about. Finally, when he had convinced himself that yes, here was a marker chromosome in a suspect cell line, a marker that he recognized -- but only when he was dead sure -- he would smile broadly and say, "Well, lookie here."

With those words, Flandermeyer had helped Nelson-Rees finger the Russian cell cultures as HeLa contaminants in the fall of 1973. Like a dogged assistant detective, he had pored over mug shots of the Russian chromosomes for hours until -- "Well, lookie here" -- he found all four of HeLa's known marker chromosomes. The Russian case had been their first practical application of the banding technique. Now, a few months later, Flandermeyer sat in his office, methodically sorting through the mug shots of two more cell lines of questionable identity. One was a line of breast cancer cells called HBT3, HBT for human breast tumor; the other was a culture of human embryonic kidney cells designated HEK.

Nelson-Rees had become suspicious of the two cultures several weeks earlier after walking in on a conversation between Adeline Hackett and Trudy Buehring, two researchers at the cell culture lab who worked independently of his cell bank. Hackett and Buehring were talking about this puzzling culture they had come across while studying the common structures of breast cancer cells. Spread out on the counter in front of them were photographs of cells from a number of established breast cultures. The pictures, taken through an electron microscope, showed that one of the cell lines, HBT3, had none of the features found in the three or four others.

"They just don't look like breast cells," Buehring was saying. "They don't look like normal breast cells, and they don't look like cancerous breast cells."

"Isn't that strange?" said Nelson-Rees. Then he remembered that he too had seen something funny in HBT3, almost a year earlier, when a California researcher under contract to the institute sent him a sample for analysis and deposit in the bank. Flandermeyer had not yet joined the staff and the only means of checking chromosomes was the conventional staining. Nelson-Rees's examination confirmed that HBT3 cells were of human origin, and that was as far as it had gone.

Except that he had seen an oddly shaped chromosome. His conventional staining had shown no detail, but the outline of the thing clearly resembled two little ears. Someone had called them Mickey Mouse ears. And come to think of it, he had seen a somewhat similar set of mouse ears in another cell line three years before that, in 1970. They were in a line of embryonic kidney cells called HEK. Maybe this oddball cell of Hackett and Buehring's doesn't look anything like a breast cell because it is actually from a kidney, thought Nelson-Rees.

He hurried off to pull the old sample of HEK out of the deep freeze and get Flandermeyer working on a comparison of the two cultures. The banding technique would show for certain whether the ears in one were the same as the ears in the other.

***

"Well," said Flandermeyer after an eternity, "lookie here."

The banding patterns of the mouse-eared chromosome in HBT3 precisely matched the patterns of the one in HEK. But that wasn't the end of it. There was another abnormal chromosome in HBT3, a new marker that Flandermeyer had never seen before, a long and boldly striped thing whose twin brother was smiling out at him from HEK as well. Two markers in common made an even stronger case that these were the same cell line. But ... what's this? Something else? Uh-oh, lookie here: four more weirdos that Flandermeyer recognized instantly.

Nelson-Rees was amazed. Buehring and Hackett were flabbergasted. It appeared that HBT3 and HEK had been mixed up, as Nelson-Rees had suspected, but neither cell line retained its original identity. Both cultures were now HeLa, actually a previously unknown substrain with two new markers, but HeLa just the same. Instead of a cancerous breast culture and a normal kidney culture, what they had stumbled onto -- and what many investigators interested in breast cancer and kidneys were no doubt wasting their time studying -- were those familiar cervical cancer cells of Henrietta Lacks.

They were still reeling two weeks later when Flandermeyer emerged from a long session of mulling over mug shots to deliver more shocking news. Cell line HBT39B, yet another culture of supposed breast cancer cells that had been sent in for a routine check, also displayed the same six marker chromosomes.

It was like that point in every horror movie when the characters know there's no escape. True, there were no Saint Bernard-sized lumps of HeLa cells blocking the exits and gnawing through the telephone wires, but this horror story was for real.

First the Russian cell lines and now this. Three cultures from American scientists, cultures effectively selected at random, all taken over by the runaway cells of "our lady friend," as Nelson-Rees had started calling Henrietta Lacks. It reminded him of a remark made a few years earlier by a group of Johns Hopkins researchers who were marveling at HeLa's tenacity. "HeLa," they had written, "if allowed to grow uninhibited under optimal cultural conditions, would have taken over the world by this time."

The director of some other cell bank might have thrown out every culture of HBT3, HBT39B, and HEK, and left it at that. But to Nelson-Rees it didn't make any sense to merely note the problems, keep the cell bank pure, and let the calamities unfold elsewhere. Being the perfectionist that he was, Nelson-Rees couldn't stand the thought of all that error and confusion, not to mention the time and money undoubtedly being squandered by HeLa's victims. Being the keeper of the cells for the institute, the man armed with the latest techniques of cell identification, he felt almost duty bound to sound the alarm, to track down the fugitive cells of Henrietta Lacks, and to set things straight.

This was the quiet beginning of the crusade. There was no formal declaration. Nelson-Rees never called the troops together to say, "The war is on," and map out strategy. What he did, quite simply, was throw the entire operation into high gear. He began by tracing the path of the first breast culture. HBT3 had come to Oakland from a researcher at the California State Health Department, who, Nelson-Rees discovered, got it from a scientist at the Centers for Disease Control in Atlanta, who got it from Robert Bassin, an institute scientist, who originated the cell line in his Bethesda laboratory. Nelson-Rees wrote to all three. "Fully realizing the embarrassment to the originators of these cell lines and/or to the investigators from whom I obtained them, I would be very pleased to discuss this matter in detail with you to get at the source of this contamination, if indeed this is what is shown," he wrote. "I welcome and in fact must insist on further analysis of these and 'related' cell lines . . . ." It was like a note from the school nurse informing the parents that little Darlene had VD, and it drew the kind of reaction you'd expect.

Bassin was the first to respond. Being a careful scientist, he was well aware of the threat of HeLa contamination, which is why he had kept HeLa and all other human tumor cells out of his laboratory when he established HBT3 in 1972. Furthermore, his lab was housed in building 41, the institute's Emergency Virus Isolation Facility.

Building 41 was cut off from the world. No agent from the outside environment could leak in to jeopardize the purity of the experimental conditions there; none of the viruses or other nasty things they handled inside was able to leak out. There were no windows in building 41, no doors that opened without special clearance. The flow of air throughout the building was carefully controlled. According to institute legend, the high security so impressed a young medical intern who worked there one summer that it later moved him to write a science fiction novel about a deadly germ from outer space, The Andromeda Strain. In fact the book's author had never set foot inside building 41, but the facility's hermetically sealed atmosphere inspired those kinds of stories. It also made Bassin confident that Walter Nelson-Rees didn't know what he was talking about. Bassin telephoned to tell him so.

First of all, Bassin argued politely, it is very difficult to prove scientifically that two things are the same. It's simple to say they are different, of course. All you have to do is find some characteristics they don't have in common. But the fact that a few funny-looking chromosomes appear in both HBT3 and HeLa doesn't prove they are the same cell. Bassin questioned Nelson-Rees about this method of examining banded chromosomes. Could he be sure the markers matched identically?

Quite sure, said Nelson-Rees, adding that HBT3 was also carrying the A type of the G6PD enzyme, the type carried by HeLa cells.

That didn't prove a thing, Bassin came back, since the woman whose tumor established the HBT3 line was of northern Mediterranean extraction -- Greek or Italian. Although it happened very rarely, type A had been known to show up in these populations. And even if the culture of HBT3 in Oakland truly was HeLa, wasn't it perfectly possible that the ones he worked with in Bethesda were bona fide? Maybe the California scientist who sent the cells in to be checked had contaminated them with HeLa in his own lab. Or maybe it happened in Atlanta.

Perfectly possible, Nelson-Rees agreed, which was why he needed to examine a culture from Bassin's personal supply.

The following day, a shipment of breast cancer cells took United Airlines flight 57 from Washington, D.c., to San Francisco. A messenger delivered them to Nelson- Rees, who passed them on to Flandermeyer for analysis. Nelson-Rees also shipped a sample to Ward Peterson in Detroit for the G6PD testing. As he waited for the results, he continued writing letters, making phone calls, notifying and debating researchers who were connected with the three contaminated cell lines. Like Bassin, none of these researchers had been working with HeLa, or so they claimed. None put much credence in the method of chromosome banding. And those who conceded there might be a problem unanimously pointed the finger elsewhere.

Ernest Plata, the man who had originated HBT39B, the other breast culture, had considerably more in common with Bassin than any of the others. He had started his cell line a few months after Bassin had got HBT3 growing. And he had done it just down the hall from Bassin's lab, in the institute's windowless fortress.

Well, well, what a coincidence, thought Nelson- Rees. It was obvious to him that a HeLa culture was running around building 41, masquerading as at least one other cell line. And while the HeLa cells couldn't leak out the carefully monitored vents or the air-locked doors, they were far from trapped. Every so often Bassin, Plata, or perhaps some other unwitting accomplice of Henrietta Lacks would wrap up a few samples of contaminated cells and mail them off. Nelson-Rees would have to have the test results to be sure, of course, but it looked as though the National Cancer Institute was distributing HeLa cells, under various false names, all around the country.

The last member of the triad, HEK, Nelson-Rees could not trace back to its source. It had been established ten years earlier by a laboratory that no longer existed. Although it had been widely used in research, there were no records of the donor's race, sex, or any other characteristics that could have been checked by his chromosomal and biochemical techniques. The earliest cultures he could find were in the hands of researchers at Pfizer Laboratories in Maywood, New Jersey, who had had them since December 1964. He asked for a sample.

By February 1974, two months after Nelson-Rees and colleagues had found the first indications of this three-way contamination, they had called in and analyzed two or three specimens of each cell line. In every one of them, Peterson had detected type A G6PD. In everyone of them, including Bassin's personal supply of HBT3, Flandermeyer had found the marker chromosomes -- the four traditional markers as well as Mickey Mouse and the one with the bold stripes, which they had named the Zebra.

Because this strain of HeLa had developed two markers not present in other known HeLa cells, Nelson-Rees and Flandermeyer decided that it must have been evolving on its own for some time, perhaps in an isolated environment such as an institute laboratory. Personally, Nelson-Rees suspected that this variant of HeLa had first contaminated HEK, the old-timer, and then gone on disguised as HEK to spoil HBT3 and HBT39B. But such reconstructions were a secondary concern at the moment.

It was time to get the word out. In the few months it had taken to track these three lines, they had come across two other popular cultures that were contaminated with HeLa -- one a line of prostate cells, the other a culture of liposarcoma, a tumor of fatty tissue. There seemed to be HeLa contaminants everywhere they turned, though few people aside from the researchers Nelson-Rees had contacted had any hint of trouble. Nelson-Rees was still pushing, unsuccessfully, to publish the very first HeLa mix-up they had uncovered, the case of the Russian cells. Originally he had wanted to use the Russians' misfortune as a warning that HeLa might yet be alive and lurking around American laboratories as well. That warning was now well behind the times. These latest findings about five American cultures demanded some kind of all-out emergency alert.

"Anybody interested in working with characterized cell lines of bona fide purity of origin would be interested in this article," he wrote to Philip Abelson, editor of the journal Science. "Knowing that cell cultures presumably derived from human embryonic kidney, human breast carcinoma, human prostate tissue, and human liposarcoma cells are indeed derived from a human cervical carcinoma would certainly change the course of a number of research projects now in progress, and alter the interpretations in many publications already in existence involving these cells."

Science sent his manuscript to their technical reviewers, one of whom criticized Nelson-Rees's writing style, though he said he was sure that the findings were correct. The other said, "The main message of this paper is extremely important: that a surprisingly high proportion of cell lines are not what they are purported to be." In view of the rapidly increasing use of cell lines in research, the reviewer added, it is vital that Nelson-Rees's message be widely disseminated. From those comments, the editors at Science somehow decided that the report didn't quite meet publication standards. No thank you, they wrote back to Nelson-Rees.

How's that? Not interested in publishing the news that five cell cultures widely used in cancer research today are not what they're supposed to be? Was it happening again, just as it had with the Russian cells -- this dead silence, this dumb stare, this gaping lack of interest in what he knew to be findings too incredible to ignore? Not this time. No, this time the sheer shocking momentum of his results couldn't be stopped. Rumors were already circulating. At the urging of several scientists and friends who had heard them, Nelson-Rees re-submitted the manuscript to Science.

It was then that Bob Bassin did a brave and unusual thing. After studying Nelson-Rees's banding data, and having performed a few tests of his own, Bassin conceded that his HBT3 cells might well be HeLa. This was no private confession. Bassin wrote to twenty researchers around the world to whom he had sent samples of his cell line, informing them of the bad news and asking that they send copies of his letter to anyone they had shared the cells with. Among other things, it was a graphic illustration of how far such an error might perpetuate itself.

Bassin also sent a copy of the letter to Nelson-Rees, who forwarded it to the people at Science with a note saying, "You will, no doubt, appreciate the need for our publication." This time they appreciated it. The manuscript was approved and rushed into type. Nelson-Rees dictated a revised introduction and checked the galleys by telephone.

A few weeks later he was in Miami, attending the annual meeting of the Tissue Culture Association, when Science mailed out the issue carrying his report. A desperate fellow approached him at the pool of the Hotel Deauville, where the conference was being held. The man looked deeply troubled, as if he had just learned that tomorrow after breakfast the universe would blink out of existence.

"As I left the lab today," the man said blankly, "I saw the Science article. I just couldn't believe what I read."

Nelson-Rees had no idea how to respond. The lost soul turned and wandered off behind the lounge chairs and umbrellas.

Suddenly people were stopping him in the hallways to ask about chromosome banding and dropping by the dinner table to check a point about sterile procedures. The next day the place was positively buzzing about HeLa contamination, and the conference organizers asked Nelson-Rees to deliver an impromptu talk on his work. He didn't hesitate.

At last, he had got someone's attention.
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"Goof" Costs 65 Years to Infinity of Cancer Research: Excerp

Postby admin » Wed Mar 09, 2016 3:24 am

8: Spreading the Word

The daily press knew how to handle this story. So what if Science buried Nelson-Rees's report in the back pages under the stodgy title "Banded Marker Chromosomes as Indicators of Intraspecies Cellular Contamination." The newspapers, properly horrified, played it on page one with headlines more to the point:

CANCER WAR SET BACK
GOOF COSTS 20 YEARS OF RESEARCH

A line of human tumor cells used by laboratories around the world for more than 20 years may have invalidated millions of dollars worth of cancer research, according to a scientist's report .... As a result, says the author, Dr. Walter A. Nelson-Rees, checks are in order for dozens of laboratories engaged in cancer research. -- Los Angeles Herald Examiner


DEAD WOMAN'S CANCER CELLS SPREADING

Dr. Walter Nelson-Rees, one of the most experienced cell biologists in the world ... has reported that many cell lines are by no means what they are thought to be by the laboratories handling them. -- Miami Herald


A SHOCKER FOR SCIENTISTS

"The main situation has probably existed for years," said the main author of the report, Walter A. Nelson-Rees, a highly respected researcher. ... Nelson-Rees said the contaminating potential of the HeLa cells is well known, but that sufficient precautions against it have apparently not been taken. -- San Francisco Chronicle


All this publicity made no sense to a number of scientists. Why was Nelson-Rees taking bows now when Stan Gartler had dropped the original bomb in 1966?

Part of the reason was that Nelson-Rees's paper was printed in Science, one of the few technical journals that nonscientists, particularly reporters, find accessible. One section, prepared by the journal's news staff, was actually written in English, and in the 7 June 1974 issue, the section carried a story that translated Nelson-Rees's article beautifully. "If Nelson-Rees is right," wrote Barbara Culliton, "a lot of people may have been spending a lot of time and money on misguided research. If, for example, you are studying the properties of human breast tumor cells, hoping to find features that distinguish breast cells from others, and are, all the while, dealing unknowingly with cervical tumor cells, you've got a problem." That was plain enough even for a newspaper reporter to understand, and to embellish and bang out for the morning edition.

But what really made Nelson-Rees a media star was the dramatic background of his shocking results: "The War." In Gartler's day, HeLa contamination had been the dirty little family secret of the tissue culture crowd. Its broader impact was not obvious. In 1974, however, "The War" had been officially declared and raging for several years. Everybody knew that the nation's most brilliant medical experts were at this very moment working feverishly against the scourge of cancer. It was a national priority.

Nelson-Rees's message made this large and serious effort seem a little silly. Sure, the institute was spending millions of dollars sending its brave recruits over the top against the enemy. But it turns out our boys were shooting with blanks!
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