A Conspiracy of Cells: One Woman's Immortal Legacy and the M

Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:04 am

10: Provenance

Walter Nelson-Rees is driving away from Oakland, toward the East Bay hills and Orinda. Although the passenger has requested no tour, he is from out of town and Nelson-Rees cannot resist. "This street we're on was the old Broadway of Oakland. That's old Tunnel Road over there. The freeway on our left was built with all the fill they dug out of the hill up ahead to make the Caldecott Tunnel. On the right here is a new sports complex, a recreation area I guess you'd call it, that is in the process of being constructed .... "

He turns the car back in the direction of Oakland and heads downtown.

"This church over here on Castro Street used to sit one block that way on Brush Street. They actually lifted the entire thing and moved it for the freeway."

The visitor begins to speak, but -- "Originally, I think it was Greek Orthodox. Then some kind of congregational church, and then a synagogue. I believe it's Methodist at the present time. Though I'm not sure of that .... "

The man's incredible, thinks Nelson-Rees's prisoner, he even spells out what he doesn't know.

"These are all original little Victorian houses. Bret Harte lived and worked in this area. He had a place with his stepfather here on Fifth and Clay, his stepfather who was the model for the character of Colonel Starbottle in his short story "The Romance of Madrono Hollow." Do you know Bret Harte, the writer?"

The passenger hesitates, then nods, wondering whether a nod will bring momentary relief or only encourage his captor.

But it makes no difference. Nothing can stop Nelson- Rees now. He is free associating in high gear, interrupting himself at every turn, improvising around the one tune his brain is always humming: Origins. Histories. Where things came from and what they used to be.

It wasn't just buildings and roadways. He read postmarks, for instance, as if they were clues to buried treasure. If a letter arrived in his office without one, he would take out his red felt tip pen, circle the stamp boldly, and write "NO CANCELLATION!" across the envelope before filing it away. It really bothered him not to know where and when the thing had been mailed.

On napkin strips and pages tom from scientific journals, he often wrote up little summaries of conversations he'd had. Some of his notes of telephone calls even specified that he'd been called collect. These along with letters, telegrams, memos, conference agendas, lab notes, articles, and scrawled copies of questions he had asked speakers at certain meetings -- as well as the speakers' replies -- all of these were stuffed into thick, black notebooks that filled the bookshelves in his office. As one of Nelson-Rees's close friends once explained, "When Walter says something to anybody, he's got a piece of paper with his name on it, with the date, and what was said. It's there. You can criticize it. You can impugn it. You can say it's a lie. But it's there."

It was the same with his interest in art. Provenance is the term for a painting's history, and until he knew the provenance in full detail, Nelson-Rees felt he couldn't appreciate the work. When was it painted, where was the artist working at the time, who were his teachers, was it ever exhibited? It was probably under the force of this compulsion in 1953 that he had changed his very name, from Rees to Nelson-Rees, thereby specifying both parents, his own provenance. And so it was with cell cultures. If you didn't know everything about the cell line you were experimenting on, especially its true identity, then you didn't know anything.

So it was probably inevitable that Nelson-Rees would publish a second list of HeLa casualties. As it turned out, though, there were other good reasons.

Since the first article, the one that had "indicted" five cell lines as HeLa, he and Flandermeyer had discovered eleven more HeLa-contaminated cultures. When Nelson-Rees told the people who established these cultures that the cells were spoiled, many simply refused to believe him. It was not enough to deliver the bad news to these researchers and hope that, like Bob Bassin, they would do the honorable thing. As each of the eleven turned up, it became increasingly apparent to Nelson- Rees that he would have to warn the world himself through another published list.

***

SH3, the purported breast cancer culture established by Gabriel Seman at the M.D. Anderson Hospital, was one such case. Seman consulted a karyologist at Anderson who disagreed with Flandermeyer and Nelson-Rees's judgment that SH3 had HeLa chromosomes, the conclusion with which Miller had concurred. Seman also changed his story about the origin of the culture. After examining a photograph in his files, he concluded that the woman who donated the cells was not Caucasian as he had originally reported, but Mexican. Since the rare type A enzyme occasionally appears among interracial Mexican-American populations, according to Seman, this meant SH3 was not necessarily a HeLa culture.

Having satisfied himself that SH3 was a bona fide breast culture, Seman saw no reason to curb its use or warn colleagues. While Nelson-Rees was preparing his new list, which would blow the whistle on SH3, the cell line was already being used by breast cancer researchers at the Memorial Sloan-Kettering Cancer Center in New York and by a group in the Soviet Union, as well as by an investigator who worked separately from Seman at Anderson. Indeed Seman would soon publish his own report in the journal Cancer, officially announcing the availability of this new breast cancer culture to the scientific community at large. In the article, he would rule out the possibility of HeLa contamination.

***

ElCo was a cell line that Nelson-Rees chose to include on his second list for similar reasons. According to oncologist Roland Pattillo, the man who established it,  ElCo was yet another breast cancer culture. In his laboratories at the Medical College of Wisconsin, Pattillo had used the cells to test the effects of certain chemotherapy drugs before administering them to the woman from whom the cells had supposedly been taken. He also studied the reaction of other breast cancer patients to this "breast culture" using a skin test something like the ones used to check for allergies. These patients showed an immune reaction to ElCo as if their bodies had already been primed against it, suggesting to Pattillo that different breast tumors might have certain characteristics or antigens in common.

The only problem, Nelson-Rees told Pattillo after checking the culture, was that ElCo was neither a breast cancer culture nor representative of Pattillo's patient. It was a HeLa culture.

Pattillo protested that breast cancer patients would never have had an immune reaction to the cells unless they were genuine breast cancer cells. He would later concede, however, that he never tested that assumption by checking patients' reactions to HeLa cells. Pattillo also explained that he knew all about the infamous tumor of Henrietta Lacks, having been an associate in the laboratory of George and Margaret Gey soon after Mary Kubicek placed that fateful bit of tissue into the roller tube. But he said he had no HeLa cells in his own lab, so contamination was impossible.

Nothing in his lab was labelled HeLa, that was true. But he had been experimenting with another breast culture, Bassin's HBT3, and apparently HBT3 had dropped in unexpectedly on some of the other cell lines in the neighborhood. Nelson-Rees and Flandermeyer's analysis showed that ElCo contained the very same banded chromosomes as did HBT3 and its HeLa-contaminated cousins: the four standard HeLa markers and the newer pair, the Mickey Mouse and Zebra.

Pattillo just didn't believe it. He kept working with  ElCo as if it were a breast tumor culture and kept publishing results. "That's what really got the wheels rolling around here," Flandermeyer recalled. "That's the kind of thing that got Walter all fired up and publishing new lists and writing letters to editors."

But what worried Nelson-Rees more than Pattillo's reaction was the fact that HeLa cells were still circulating in their old HBT3 disguise. HBT3 had been on the first list in 1974. For the last two years he had been making as much noise as he could about HBT3 and the four other HeLa-contaminated cultures he had originally stumbled upon. And yet here was Pattillo claiming he had no HeLa in his lab -- although, yes, he did use a little HBT3 now and then.

***

Unfortunately, HBT3 was not the only active alumnus from the first list. Nearly everyone, it seemed, was still using MA160, the bogus prostate cell that by the mid-1970s had helped researchers waste an estimated $200,000.

MA160 was developed in 1966 through the collaboration of several scientists and a man named Monroe Vincent, a partner in the biomedical supply firm of Microbiological Associates, Inc. The scientists provided some of the technical expertise; Monty Vincent personally contributed the cells, a lump of tissue that had been taken from his own prostate. The lump was biopsied by his doctor who was concerned that it might be malignant. Fortunately, it turned out to be benign.

It also turned out to be a good source of culturable cells, or so it seemed. The cells from Vincent's prostate languished in the laboratory for a few months and then, touched by the miracle of "spontaneous transformation," were reborn as a bunch of frantic cancer cells. The first long-lived line of prostate cells, MA160 became a bestseller.

As it happened, though, the cells were exactly the opposite of what they were supposed to be. Not male, but female. Not prostate, but cervix. Not from a white donor, but a black one. MA160 was arguably the most tasteless of HeLa's practical jokes.

Stan Gartler was the first to nail MA160 as HeLa in 1968. He added it to his list of the original 18 HeLa-contaminated cultures based on its having the A type of G6PD enzyme. Vincent and his co-cultivators, however, dismissed the finding in their first official description of the cell line in Science in 1970. They theorized that Vincent might have had Negro ancestors from whom he inherited the rare, black enzyme. They added that the cells contained Y chromosomes, proof they had come from a male donor.

In 1973, however, Ward Peterson, Nelson-Rees's regular partner in Detroit, reported that he could find no Y chromosomes even in the earliest samples of MA160 available. Furthermore, having tested Vincent's own blood, he could say without a doubt that whatever exotic ancestry Vincent claimed, he had ended up with type B G6PD, the type expected for whites. "His" cell line was certainly not his anymore, but that of a black woman. In 1974, Nelson-Rees and Flandermeyer found that MA160 not only had the type A enzyme and was missing its Y chromosome, it also displayed banded marker chromosomes identical to HeLa's. Another group that had been working with the culture, a team at the Anderson Hospital, no less, soon came to the same conclusions.

It wasn't long before Mukta Webber, the Colorado cell biologist, reported that MA160 failed to produce prostatic acid phosphatase, a chemical normally manufactured by prostate cells. Webber also reported that despite its unprostatelike behavior, MA160 was still a popular research culture, describing and giving references to the six recently published "prostate" studies based on the cell line.

But that was not the end of MA160. Early in 1975 a West German scientist named Frederick Schroeder announced the establishment of a new prostate culture at a conference in Italy. Nelson-Rees, who happened to be attending, heard Schroeder describe the new cell line, EB33, as quite similar to the prostate culture MA160, which Schroeder also had been studying in his lab. In certain respects, EB33, like MA160, behaved strangely, according to Schroeder. Nelson-Rees began asking questions. A few months later, having analyzed several samples of EB33, he wrote Schroeder that his new prostate culture was definitely MA160, which was definitely HeLa.

And still MA160 and its alter ego endured. Many researchers went ahead and worked with EB33 just as others had continued to use MA160. Some, including Schroeder, used both cultures in their experiments, presumably to increase the validity of the observations. Here it was again, Nelson-Rees's nightmare come true. Researchers were not only using MA160 as if it were a bona fide prostate culture, they were exposing all the other cell lines in their labs to a HeLa culture in disguise. Which was why in addition to warning people away from the eleven newly spoiled cultures like EB33, ElCo, and SH3, Nelson-Rees decided his second list would have to remind them of the contaminated cultures he had publicized earlier.

In fact he decided not only to repeat the lines he indicted in 1974, but also to quote every report he could lay his hands on that branded any cell culture HeLa. Among others, he cited Gartler's original findings, recent studies by Ward Peterson in Detroit, and a paper soon to be published by researchers at the American Type Culture Collection. In the manuscript he submitted to Science accompanying this second list, he explained how widespread "secondary contamination" had become and admonished his colleagues, "HeLa by many other names can spell trouble."

The list, published in January 1976, was an impressive and distressing inventory. More than forty different research cultures had been commandeered by HeLa. If each culture had cost science a quarter-million dollars, as had the few for which damage estimates could be made, this list represented $10 million in losses. The evidence against each culture, organized in a detailed chart, was overwhelming too. Not only were these cells identical to HeLa on the basis of their C6PD enzymes and marker chomosomes, but many had also been tested for the enzyme PCM and for a few more biochemical traits called HLA antigens. These all matched HeLa's characteristics.

And, oh yes, there was one other piece of information under each cell line in Nelson-Rees's second list: a researcher's name. Any scientist who had supplied him with a culture that turned out to be HeLa was immortalized in what came to be known as Nelson-Rees's hit list. No one considered this an honor. A few felt deeply betrayed and never spoke with him again. And the ranks of those who thought Nelson-Rees was just out to make a name for himself grew considerably.

Jim Duff and some of the other bureaucrats at the institute wanted to know why he had to name names. "You keep pushing people," Duff complained. Why, they wondered, couldn't Nelson-Rees simply give the sample's "passage level" -- the number of times it had been grown out, cut up, and transferred to new flasks, which would give a relative indication of how close it was to the original culture-or identify it in some other way, and leave off the scientist's name?

Leave off the name? That, to Nelson-Rees, was like -- well, like asking the Post Office to forget about the postmark. Why not take down all the road signs and let drivers fend for themselves? Why not ask the telephone company to publish just the phone numbers and have people guess to whom they belong? To leave off the names would defeat the whole point of his list. He was trying to be as precise and complete as possible, to specify where the HeLa-contaminated cultures had come from and, by implication, where they had not come from. After all, he hadn't tested every sample of these cell lines. Not yet.

"After the 1974 paper," Nelson-Rees explained to Science News magazine, "some researchers analyzed cultures they had been using of the same type we 'fingered' and found them to be bona fide bladder carcinoma cells, or whatever. Therefore, the source of these cultures becomes an important piece of data. I felt obligated to state from whence these cultures came and let the other shoe drop where it may."

Of course, Nelson-Rees must have known that naming names was, as the magazine put it, "an action sure to be interpreted by some as unfriendly." But maybe he figured that in order to make any progress, the second list had to be more combative than the first.

"At this point," he told Science News, "I'm going to go hide."

Naturally, he did nothing of the kind.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:06 am

11: Another Run-in With Relda

Nelson -Rees was browsing through the April 1977 issue of the Journal of the National Cancer Institute when he caught the scent of something mildly suspicious on page 863.

Now some people would have said he went looking for bad news, especially after that hit list with the names on it. But the way he saw it, he just wanted to make sure the reports of good news were genuine. And starting on page 863 of the institute's journal there appeared to be very good news indeed, a report of some promising findings about lung cancer.

Richard Akeson of UCLA medical school had found several specific antigens associated with the cells in a lung tumor culture. Antigens are like biochemical dog tags, tiny nameplates on a cell's surface that can be recognized by the body's immune system, though in the case of cancer the system seems unable to successfully attack even when it recognizes the enemy. While the discovery was preliminary, Akeson wrote, such tags might eventually help doctors diagnose this particular kind of lung cancer more effectively, perhaps earlier, and help them monitor its progress. Furthermore, though Akeson didn't actually mention it, there was always the hope that if tumor-specific antigens could be found for certain cancers, they might serve as signposts to be followed by cancer- killing drugs or other selective assaults to be developed in the future. Such was the promise of an antigen found to be unique to a particular kind of tumor cell.

Nelson-Rees knew from experience, however, that antigens specific to a certain type of cell -- and only that type -- were practically impossible to find. And, of course, finding a unique antigen wasn't enough. You also had to be certain of the tumor cell's identity in the first place, so that later when you saw that antigen on the cells of an unidentified culture you could say, for example, aha, we must have a dishful of Russian bladder cancer here. Given the proclivities of biologists to mix things up, Nelson-Rees figured, any tissue-specific claim was worth a closer look.

According to the "Materials and Methods" section of Akeson's report, the lung tumor cell line was called 2563 and had come from Litton Bionetics, Inc., in Kensington, Maryland. Following that bit of information was a footnote that directed Nelson-Rees to the bottom of the page. There amid the fine print, quite unexpectedly, he bumped into none other than Relda Cailleau. It seemed that 2563 was another name for MAC-21, the lung culture that had triggered the hotel lobby screaming match in Montreal two years earlier. Despite Cailleau's assurances at the time that MAC-21 was not HeLa, the matter was left very much unresolved in Nelson-Rees's mind. Now that someone was basing important new claims about lung cancer on this twenty-year-old culture, he decided the question had to be settled.

He ordered samples of 2563 from Litton Bionetics that summer, and by September the verdict was in. According to Ward Peterson's analysis, the culture's G6PD enzyme was type A, the same as HeLa's. Although Cailleau had taken the original MAC-21 cells from a fifty-three- year-old man, Nelson-Rees and Flandermeyer saw no male Y chromosomes. And in nearly every cell, they found abnormal chromosomes whose shapes and banding patterns matched three of HeLa's best-known markers. Nelson-Rees wrote a detailed letter to John C. Bailar III, editor of the Journal of the National Cancer Institute, asserting that 2563 had been contaminated by HeLa and was not at all characteristic of lung tumor cells. He asked that the letter be published as soon as possible, saying his results were "an important addition to the increasing knowledge of what does and does not constitute organ-or tissue-specific antigens."

Three months passed with no word. Nelson-Rees called the journal, and someone there explained they had sent copies of his letter to Akeson and Cailleau so that they could write rebuttals to be published along with his letter. Akeson's reply had come in, a cautious concession that the "lung cancer cells" were apparently HeLa, but they were still waiting for Cailleau's.

Convinced as ever that the cells were pure, Cailleau had pulled out the one remaining ampule of MAC-21 in her possession and had it analyzed by Sen Pathak, a chromosome banding expert at the M.D. Anderson Hospital, and Michael Siciliano, the local enzyme man. She was "disappointed" as she later described it, when Pathak reported seeing HeLa's marker chromosomes in MAC-21 and Siciliano said not only was the G6PD HeLa's type, but thirteen other enzymes he tested matched as well.

Disappointed, but undaunted, Cailleau still insisted that MAC-21 had been a genuine lung cancer cell for many years, and if it was HeLa now, well, that was somebody else's doing. She explained to her colleagues, as she would afterwards to anyone who asked about MAC-21, that she had shared the cells with other researchers in San Francisco before coming to Houston. When she moved she took with her two different batches: one of live samples from those researchers and another of frozen samples that only she had used. Then in the early 1970s some fool technician had failed to keep the freezer stocked with dry ice, and she lost seventeen years' worth of cell cultures, including the bona fide MAC-21 cells. Her only remaining samples were those that she had had no direct quality control over.

"The problem," Cailleau said, "is that often contamination of cells is a secondary or a tertiary thing done by associates or technicians, and the originator knows damn well he or she didn't contaminate them. In my hands, there never has been a HeLa contamination." In other words, yes, the MAC-21 cells she had now were indeed HeLa, but it wasn't her fault. The same was true for the samples she had sent a few years earlier to a variety of colleagues, including one who must have passed them on to Litton Bionetics under the name 2563, some of which had ended up in Richard Akeson's lab.

All this Cailleau explained in a letter to Bailar at the Journal of the National Cancer Institute in January 1978. Painful as it must have been, she also telephoned Nelson- Rees -- collect -- and laid out the whole mess for him. She sounded uncharacteristically subdued, Nelson-Rees thought, even a bit sad. As pleased as he was to hear his suspicions confirmed, he kept his I-told-you-so's to himself.

Still, it would be another six months before anyone but Nelson-Rees, Akeson, Cailleau, and Bailar had any inkling that MAC-21 and 2563 were not what they were advertised to be. Scientists who had read Akeson's report on lung tumor-specific antigens would have had an entire year to follow his lead without any indication it might be a dead end. Finally Nelson-Rees's letter appeared in the journal's June 1978 issue. It was followed by a letter from Akeson who said that based on Nelson-Rees's work "it seems reasonable to presume" the cells he had thought were from a lung tumor were actually HeLa cells. He added, however, that some of the antigens in these cells did seem to resemble those seen in genuine lung tissue, concluding that further studies were needed to sort through the confusion. Following Akeson's letter was a one-paragraph statement from Cailleau conceding that the single culture of MAC-21 she had checked was HeLa. "I am certain that the original MAC-21 was obtained from a mucoid adenocarcinoma of the lung and remained uncontaminated for several years," she wrote with a trace of controlled testiness. "The addition of HeLa cells to our cultures occurred later and the transfers were carried out by several people."

Around the time of this embarrassingly public cell mix-up, Nelson-Rees received a draft report from a microbiologist at the National Cancer Institute. It was an extensive review of breast cancer cultures available to experimenters, and, as Nelson-Rees was so experienced in telling good cells from bad, the microbiologist asked him to critique the report before it was submitted for publication. Oddly enough, there were several cultures listed whose technical descriptions were missing. The cover letter explained these were Cailleau's cultures, and Cailleau had asked that all information on them be deleted from any copies of the draft that Nelson-Rees might see. Nelson-Rees was amazed-first that Cailleau would have the gall to dictate the handling of someone else's manuscript and, second, that the author would knuckle under. When Nelson-Rees called to find out more, the microbiologist said that Cailleau had been "adamant on this point; abusive, in fact."

Obviously Cailleau had been more than just miffed by her latest dealings with Nelson-Rees. She was determined to make the MAC-21 incident the last time he would ever stick his nose into one of her cell cultures.

***

A few weeks later, the Tissue Culture Association asked Nelson-Rees to evaluate a stack of abstracts, or summaries, of research reports submitted for the association's upcoming annual meeting. He found one of the abstracts particularly interesting. It described an unusual sample of breast tumor cells that exhibited a surprising number of characteristics similar to HeLa's. In fact, many of its chromosomes looked like HeLa's banded markers, the authors said. Yet they knew for certain the cells were not HeLa because they had come directly from the patient -- out of a pleural effusion, a sample of liquid taken from the patient's chest cavity, to which the tumor cells had spread. The cells in the effusion were analyzed immediately; they had not sat around the lab and had a chance to become contaminated. The implication was that in this case the standard techniques of identifying a HeLa cell might well have led to a false diagnosis. These bona fide breast cancer cells might have been wrongly branded as having been overtaken by HeLa.

The abstract had been submitted by three scientists at the Anderson Hospital: Sen Pathak, Michael Siciliano, and Relda Cailleau.

Nelson-Rees almost laughed. He doubted the claim, of course, but was intrigued at the same time. In part because he wanted a chance to examine the supporting data, he recommended that the report be accepted and presented at the meeting. He then wrote to Pathak, the karyologist, asking to see photographs of these curious chromosomes.

When the photos arrived two months later, eagle-eye Flandermeyer looked and looked but he couldn't see any known HeLa markers among the banded chromosomes. Nor could Nelson-Rees. They sent copies to K.S. Lavappa, the chromosome expert at the American Type Culture Collection, who wrote back that there certainly were some strange-looking chromosomes in these cells, but none of them were HeLa markers.

Two weeks before the annual meeting, at which Pathak was scheduled to discuss this cell line, Nelson- Rees received yet another manuscript, this one from Bailar, who asked him to review it for the Journal of the National Cancer Institute. It was a much more detailed report on the very same cell line. In addition to Pathak, Siciliano, and Cailleau, T. C. Hsu, head of the Tissue Culture Department at Anderson and described by some as the guru of cytogenetics, had put his name on the manuscript, adding considerable credibility to it.

What the earlier abstract had only implied, this twenty-five-page paper announced loud and clear: that the standard tools of HeLa hunting -- enzyme tests and chromosomal analysis -- were of questionable value. What's more, it suggested that every HeLa case Nelson-Rees had ever published was now in doubt. In short, it looked like the gang from Anderson was indicting the indicter himself.

"The HeLa markers have been extensively used by investigators to identify ... cell line contamination," the authors wrote, citing references to three of Nelson-Rees's published reports, including the latest hit list. Because they found chromosomes similar to HeLa markers in this tissue sample that was clearly not HeLa, they concluded that "marker chromosomes alone are not unequivocal evidence for identifying cellular contamination."

Wait a minute, thought Nelson-Rees. From what he and Flandermeyer and Lavappa could see, there were no HeLa markers in these cells. Sure, maybe there were a few that looked vaguely similar. Maybe someone with an untrained eye might be confused. But there was not a single marker chromosome in the photographs that a qualified karyologist should mistake for HeLa's. And even if there were, Walter Nelson-Rees would never label a cell culture HeLa based solely on marker chromosomes. There were also the enzymes to consider.

But the folks from Anderson observed that twelve out of fourteen isoenzymes tested in their breast cancer cell were identical to HeLa. "The suggestion, then, that cell lines showing only three enzyme characteristics in common with HeLa be considered de facto strains of HeLa may be overly simplistic," they wrote, offering footnotes to two more of Nelson-Rees's reports.

Overly simplistic{ Nelson-Rees couldn't believe this stuff. He'd never claimed that enzymes alone are enough. But if the marker chromosomes match and you pick the proper enzymes to check, then three can be plenty. The single finding of type A G6PD, for instance, in a culture thought to be from a white donor is more than enough to tell you something is awry.

Ah, but the "HeLalike" cells reported in this manuscript had come from a black woman who carried the A form of G6PD. How convenient, thought Nelson-Rees.

As he scanned the rest of the report, he noticed the authors neglected to point out that many of the other enzyme variants they tested were common to 90 percent of the human population, including the late Henrietta Lacks, and therefore of relatively little value in distinguishing their breast cancer cell from HeLa. And by a strange coincidence the two enzymes that turned out different from HeLa's happened to be the very last two they tested, even though one of them, called PGM3, had become a standard test for HeLa, and therefore likely to be among the first checked by anyone really interested in finding out if he had a HeLa cell or not.

The handling of the enzyme data was not the only thing Nelson-Rees found suspicious. He called Flandermeyer in and they pored over the chromosomal mug shots Pathak, Cailleau, and colleagues had used to prove their case. In an hour or so they were both shaking their heads, astonished.

The mug shots were photographic composites, constructed from bits and pieces of various photos of various cells in this breast cancer culture.

When you're comparing banded chromosomes from two different tissue samples -- a known HeLa culture, say, and one you suspect may be contaminated with HeLa -- there's only one correct way to do it, according to Nelson-Rees and most other karyologists. After searching hundreds of photos you begin to recognize the same complex of markers, the same group of defective chromosomes, appearing in every one of your suspect cells. You then pick a single cell in which this group displays its banding patterns clearly, cut out the defective chromosomes, and put them next to markers from a HeLa cell you've analyzed and photographed. In a given side-by-side comparison, the mug shots must come from only two individual cells. If you have to pick one chromosome from this suspect cell and another from that in order to find ones that match those from the HeLa cell, then you're reaching.

Apparently that's just what the Anderson workers had done. After a close look at all the photographs, Nelson- Rees and Flandermeyer were convinced that the comparison photo didn't display mug shots of chromosomes from only two cells, one from the breast culture and one from a HeLa line, but from at least seven, four different breast cells and three HeLas. What's more, the HeLa mug shots appeared to have been taken from another researcher's published report, not a culture the Anderson workers had studied themselves. In fact, they had cut out and reassembled so many mug shots in order to make their comparison look convincing they had even pasted in the image for one of the breast chromosomes upside down and reversed.

So this was what Pathak was going to unveil at the annual meeting. And as if that wasn't enough, the Anderson gang wanted to publish the entire argument in the Journal of the National Cancer Institute as well. Nelson- Rees was astonished that anyone would go to such lengths to discredit him.

For the next few days he drove Flandermeyer and the others hard. He demanded a detailed analysis of the mug shots showing that they weren't HeLa markers despite the photographic sleight-of-hand and offering alternative descriptions for each chromosome. When that was done, Nelson-Rees sat down, poison pen in hand, to draft a statement he planned to deliver at the annual meeting following Pathak's presentation. When it was done, he took it to Flandermeyer, who recommended toning it down. They removed a few of the more offensive passages, including a final statement Flandermeyer felt was particularly undiplomatic. Nelson-Rees then returned to his office and wrote the venom back in.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:07 am

12: Showdown

As soon as Nelson-Rees arrived at the Denver Hilton Hotel, where members of the Tissue Culture Association were gathering for their twenty-ninth annual meeting, a friend came up and said, "The Anderson group is out to get you."

Privatization of National Cancer Policy

In February 2002, Senator Dianne Feinstein introduced the National Cancer Act of 2002. Co-sponsored by 30 bipartisan Senators, including Majority Leader Tom Daschle and Hilary Clinton, the Bill was a radically different version of President Nixon's 1971 Act that launched the National Cancer Program. The Bill added $1.4 billion to the $4.6 billion 2003 budget authorized by President Bush, extra funds coming from the new Federal cigarette tax increase, and a further 50% annual increase to 2007, reaching a grand total of $14 billion. Feinstein said her goal was to "form our new battle plan to fight cancer." The legislation was referred to the Committee on Health, Education, Labor, and Pensions, then chaired by Senator Judd Gregg.

This Bill established a national network of 20 "translation" centers to combine basic and clinical research, and to commercialize promising findings. It also mandated insurance coverage for cancer screening, smoking cessation, genetic testing, and quality care standards, while making no reference to prevention.

Regrettably, this well-intentioned Bill surrendered the National Cancer Program to special interests. The legislation was strongly criticized by survivor coalitions, headed by the Cancer Leadership Council, and the American Society for Clinical Oncology. Of major concern, the Bill displaced control of cancer policy from the public to the private sector, the federal NCI to the "nonprofit" ACS, raising concerns on conflicts of interest and special interests. Dr. John Durant, Executive President of the American Society of Clinical Oncology (ASCO), awarded the Society’s 2002 Presidential U.S. Cancer Fighter of the Year, charged: "It has always seemed to me that this was an issue of control by the ACS over the cancer agenda. They are protecting their own fundraising capacity . . ." from competition by survivor groups. Not surprisingly, the authoritative U.S. charity watchdog, The Chronicle of Philanthropy, warned against the transfer of money from the public purse to private hands. The ACS is more interested in accumulating wealth than saving lives.

These conflicts of interest extended to the personal. The Legislative Committee co-chair, DeVita, was Board Chairman of CancerSource.com, a website promoting the ACS Consumers' Guide to Cancer Drugs. As disturbing, DeVita, and Dr. John Mendelsohn, Director of the NCI's M.D. Anderson Comprehensive Cancer Center, were consultants and board members of the publicly traded cancer drug company, ImClone Systems, Inc. Mendelsohn was also a board member of Enron, besides serving on its Audit Committee; Enron was a generous and long-term supporter of the M.D. Anderson. In May 2001 television and radio interviews, DeVita expressed enthusiasm on cancer drugs that targeted “EGF” receptors. However, he failed to disclose his annual $100,000 consulting fees from ImClone which was then actively seeking FDA approval of its targeted cancer drug Erbitux. DeVita also insisted, contrary to NCI’s own data, that the overall incidence of cancer had been decreasing at a steep rate every year since 1990. In May 2002, Dr. Samuel Waksal resigned as president and CEO of ImClone. One month later, he was arrested on charges of criminal conspiracy, securities fraud and perjury, and civil damages for insider trading, and was subsequently indicted on charges of insider trading, bank fraud, forging a signature and obstructing a federal investigation.

In the September/October 2002 issue of The Cancer Journal, an article by its co-editor DeVita, “A Perspective on the War on Cancer” was prefaced by the following disclaimer: “No benefits in any form have been or will be received from a commercial party related directly or indirectly to the subject of this article.” However, as pointed out in a November 15, 2002 letter (by the author) to the Journal’s other co-editors, Drs. Samuel Hellman and Steven Rosenberg, this disclaimer was inconsistent with DeVita’s conflicts of interest relating to the CancerSource.com web site, and his ImClone consulting fees. The editors of the Journal responded that it “takes matters of conflict of interest and disclosure very seriously,” but nevertheless declined to publish the letter.

-- American Cancer Society: More Interested in Accumulating Wealth Than Saving Lives, by Samuel S. Epstein, M.D.


The following morning, the inflammable Relda Cailleau ignited at the mere mention of his name. She was at a meeting of the association's executive council, where she learned that Nelson-Rees had been elected vice-president of the association and was to be officially installed the next day -- and she couldn't contain the fire and fury.

"My god," she blurted out. "How could he have been elected vice-president?"

She told her startled fellow council members how Nelson-Rees had attacked her work and that of many well-respected scientists, how his methods were antagonistic and his motives destructive. "His approach is always, 'I've got to prove I'm right and you're wrong,'" she railed. "My God, the man is paranoid!"

Cailleau urged the council to demand his resignation, a difficult task as Nelson-Rees didn't yet hold the office. But she had made her point, several times over, and set the stage for the main event later in the day.

Nelson-Rees was set himself. Earlier he had told Bob Stevenson, "I'm really going to throw the fat on the fire." "Think about it," Stevenson answered.

Some 200 people packed into the Hilton's Vail Room that afternoon, considerably more than were originally expected for the session on mammary tissue. They had heard there was quite a show in store. Some said it might really get ugly. They came not necessarily to root for one side or the other, but -- like decent townsfolk gathering on rooftops and peering out of windows at a gunfight in the street -- just to watch.

The paper by Pathak, Cailleau, and Siciliano, entitled "Fresh Pleural Effusion from a Patient with Breast Cancer Showing Characteristic HeLa markers" was scheduled third. The audience sat restlessly through the first and second talks. Then, about thirty minutes into the program, the chairman read the title of the paper, and Pathak, the young Indian-born chromosome expert, came forward to deliver the talk. Cailleau was sitting in the audience about halfway to the back of the room. Nelson- Rees was about ten rows behind her. Siciliano had stayed home in Houston.

Pathak seemed apprehensive. He spoke a bit shakily in that British-educated, Indian accent, painfully formal and polite. Whenever he mentioned a colleague, it was always by title and full name, pronounced in one breath as if it were all a single word: "We examined chromosomes in cells obtained by DoctorReldaCailleau .... Isoenzymes from DoctorReldaCailleau's cells were analyzed by DoctorMichaelSiciliano."

Pathak described the findings in a straightforward way: These cells taken directly from a breast cancer patient showed chromosomes that were quite similar to HeLa's characteristic marker chromosomes, he said. And thirteen of fifteen isoenzymes checked were identical to HeLa's. (They had tested one additional enzyme since writing the draft report Nelson-Rees had seen.) There were no direct references to Nelson-Rees, no outright challenges to the usefulness of his techniques as there were in the written version of this talk that had been sent to the Journal of the National Cancer Institute. Still, to some members of the audience, especially to Nelson- Rees, the insinuation was clear.

When Pathak had finished, Nelson-Rees raised his hand. The session chairman, having heard that trouble was afoot, quickly called on someone else. Nelson-Rees decided he would not be denied again. When that question was answered, he simply stood up and without the aid of a microphone began to read his statement.

"As a referee of this abstract for the program committee, I questioned the use of the adjective 'characteristic' in the title, followed by 'similar' in the text. Dr. Pathak subsequently sent me a photograph of the 'marker' chromosomes stating he 'was puzzled regarding this observation.' I am puzzled as well, not with the observations but with the way they are being presented."

The chairman tried to cut him off, explaining that the presentation had gone over its allotted time.

"John," said Nelson-Rees as he moved from his seat up towards the front of the room, "I'm going to finish this."

He went on to explain that as a reviewer of the manuscript submitted to the cancer journal, he had looked over all the photographic evidence the authors used to prove their case. He said he discovered they had "fished about" in several different cells from this breast tumor to find their so-called HeLa chromosomes, a highly irregular procedure.

"We have carefully studied many different cell lines and observed a good number of marker chromosomes, but we've never had to create a collage such as has been done here in order to sensationalize a point."

The audience was aghast at such a flagrant breach of etiquette. One simply didn't use such language. Perhaps a point would need to be confirmed; maybe there was a chance for an alternative interpretation. But publicly accusing learned colleagues of fishing about for data and sensationalizing their results -- Nelson-Rees might just as well have been spoon-launching mashed potatoes at a formal dinner.

"As a matter of fact," Nelson-Rees proclaimed, "without stating so and without giving credit to the published work, the authors lifted HeLa marker chromosomes from the work of Heneen which best matched their altered chromosomes and presented them in the manuscript apparently as their own observations." He held up the Swedish journal Hereditas that contained the report he claimed they had "lifted" from and added that despite such pilferage, the chromosomes they presented are not convincing matches.

There was a sudden disturbance in the audience. Relda Cailleau was becoming agitated, muttering loudly to the people around her, but Nelson-Rees pressed on.

"Aside from the above-mentioned irregularity in scientific procedure, it appears that some normal chromosomes were erroneously classified as markers and vice versa, and at least in one instance a chromosome's image was used twice, albeit after reversal of the negative."

Nelson-Rees explained that he had sent photographs of the "so-called characteristic" HeLa markers to Lavappa for a second opinion and quoted Lavappa's written reply: "'There is no trace of the four distinct HeLa markers, which are well characterized and have appeared in many publications.'" And then it was time for the finale, the part that he had typed out in capital letters.

"I suggest you stick to pleural effusions," he said to a dumbstruck Pathak at the podium, "and leave the HeLas to us."

A momentary silence.

Pathak broke the trance and added a touch of absurdity to the scene by thanking Nelson-Rees profusely. Nelson-Rees dropped 150 copies of his statement in a pile at the back of the room and left. Suddenly pandemonium broke out over the Vail Room. There were five more speakers scheduled for the afternoon, but half the audience was rushing for the exits.

Relda Cailleau made her way over to the stack of statements, grabbed about thirty of them, and headed out. In the hallway she worked the crowd, offering copies of Nelson-Rees's attack as proof the man had no business reviewing manuscripts for technical journals (Just look at how he violated the reviewers' oath of confidentiality!) or serving as vice-president of the Tissue Culture Association (Was such rude, unethical behavior fitting for an officer of our group?).

Pathak returned to his hotel room, where he remained for most of the last two days of the conference. He would later explain his absence by saying he was deeply shaken by the wrongful accusation and public humiliation he had suffered. In fact, he was so distraught that he refused to do any research on the chromosomes of tumor cells for an entire year. Long after the incident, Nelson-Rees offered an alternative explanation: "A little dog -- or even a big one -- who has peed on the living room rug and is found out always hides underneath the couch for a couple of hours."

At the moment, however, Nelson-Rees was fresh out of quips. His feistiness was suddenly spent. He was exhausted and, he noticed, quite alone. As he moved through meetings the rest of the day, people avoided him, looked away disapprovingly when he approached, and whispered about what a disgraceful performance he had given.

One of those who attended his performance and had been especially shocked was Colonel Albert Leibovitz. A tissue culturist formerly with the U.S. Army and now at the Scott and White Clinic in Temple, Texas, Leibovitz had at one time respected Nelson-Rees's crusade to clean up the field, or so he said. Recently, though, it looked to Leibovitz like Nelson-Rees was on a maniacal rampage. His means were extreme. And what he had done this afternoon could only be described as scandalous. "It is so unethical to be a reviewer of a paper and use confidential information to tear its authors down," Leibovitz declared.

Leibovitz, of course, may have felt some special sympathy for the Anderson crew, having had a recent tussle with Nelson-Rees himself. The matter involved eight cell lines Leibovitz had established: colon, bladder, and six other kinds of cancer, all of which had apparently been taken over by a single line of colon cancer cells. Although it was not a case of HeLa contamination it was still a large mix-up, and Nelson-Rees felt it ought to be publicized immediately. When Leibovitz balked, preferring to double-check the findings and investigate the source of the contamination further, Nelson-Rees tried to publish a letter in the Journal of the National Cancer Institute, warning other scientists away from these cells. Leibovitz was furious. Certainly Nelson-Rees had been right to want to get the word out, but he was jumping the gun for no reason, Leibovitz believed, and worse, he had broken an implicit confidence -- just as he had in attacking the report by the Anderson group.

Later that afternoon at the general membership meeting of the Tissue Culture Association, it was Colonel Leibovitz who stood up and with great indignation urged that the association create an ethics committee to deal with one of its members who, at a presentation earlier in the day, "committed unscientific acts without presenting proof to support his statements." This person's verbal assault was all the more inappropriate, the colonel explained, because he had been the confidential reviewer of a manuscript covering this particular presentation.

Seated out in the audience, Nelson-Rees kept a faint, nervous smile on his face as the members discussed the need to discipline the unnamed outlaw and eventually voted to have Leibovitz draft a resolution for consideration at the next meeting of the executive board.

That night Bob Stevenson took Nelson-Rees out for a drink and a little lecture. It was another version of the friendly advice Stevenson had regularly offered: Diplomacy gets you farther. When you do something, you do it properly, or you risk losing credibility.

On the other hand, Stevenson added, he was only too glad somebody was stirring things up. He grinned his mischievous grin and said, "It's always nice to drop a turd in the punchbowl once in a while."

***

Then came the letters. In a few weeks Keith Porter, the president of the association, was hip deep in testimonials of outrage, shock, and dismay:

I am writing to express my concern about the incident that took place in Denver. Nelson-Rees has abused his privilege as a reviewer and impugned the integrity of Ors. Pathak et a1. I hope the Executive Board will treat this matter with the utmost seriousness, because I believe the credibility of the Board, as well as the good reputations of Ors. Pathak, Siciliano, and Cailleau are at stake.

I am but one of many who was shocked and offended by the serious and unethical activities of Walter Nelson-Rees.

Dr. Nelson-Rees behaved ungentlemanly and indiscreetly. He appeared to lack the objectivity and open-mindedness of character important for a good scientist.

Leibovitz's resolution also arrived, four pages documenting Nelson-Rees's crimes. "It is immaterial whether his findings are right or wrong," the statement read and urged that he be censured, stripped of his vice-presidency, and banned from reviewing manuscripts for the association.

It was all very well, all this righteous wrath, but as far as Porter could see, there was no way to satisfy it. Nelson-Rees was guilty of no malfeasance, and even if he were, he was not in office when he so distressed everyone; that was the day before he was officially installed. When the officers, including Nelson-Rees, met a few months later, Nelson-Rees agreed to apologize to Pathak in writing, but only for the manner in which he "disagreed" with Pathak's report. The president hoped that would satisfy the offended.

It didn't come close to satisfying Relda Cailleau, who wrote the president four months after the incident:

Dear Dr. Porter,

If he [Nelson-Rees] and other members of the Council consider that his apology for having "offended" Dr. Pathak is enough to close the subject, they are mistaken. His unethical conduct is not mentioned and it is the only subject of importance for the T.C.A. and for any reputable scientist .... If he cannot be removed from office or forced to resign because the T.C.A. constitution has no provision for such a contingency, he must at least be made to acknowledge his unethical conduct (which incidently is not limited to this occurrence) .... He should no longer be allowed to review articles without some safeguard to the people whose reputations he tries to destroy. The only way this can be done is to inform the membership as well as the editors of the various journals in which his comments have appeared.


Porter answered:

Dear Relda:

... Unofficially, I would urge you to forget the incident. I have heard some very abusive attacks of one scientist on another at scientific meetings and they did not become the business of the society or institution sponsoring the meeting to settle. The TCA cannot undertake to censure, punish or even reprimand its members for "unethical" behavior. As I have said before, Walter was duly elected and cannot be removed from office without going back to the membership of the Association. I do not think that you could influence the required plurality.


What amazed Nelson-Rees was that nowhere amid the outcry over his Denver recital was there any discussion of the merits of his argument, no effort made to determine whether the results reported by Pathak and company were trumped up, as Nelson-Rees had charged. They had claimed that his methods for fingering a cell line as HeLa were in doubt and, by extension, his hit lists and other reports were wrong. Well, Nelson-Rees couldn't stand being told he was wrong. And it incensed him that no one cared to consider the substance of his rebuttal.

After the Denver meeting, he sent off a stinging critique to Bailar at the Journal of the National Cancer Institute, restating and amplifying his argument. The Pathak report, he wrote, should not be published unless its" entire concept is altered." He questioned its" sincerity and merit" and accused the authors of resorting to "photographic trickery" in their presentation of the chromosome evidence. As for the enzyme data, he wrote, it may well be true that thirteen out of fifteen enzymes tested were the same as HeLa's; but two of them, the two most useful for identifying HeLa cells, were different. Hence the system holds up. No one would mistake these cells for HeLa, as Pathak and Cailleau's gang had claimed.

Like his speech in Denver, however, these criticisms were ignored. The paper, barely revised from the original version, was published in February 1979.

***

When they were asked about it years later, the Anderson researchers insisted they had never set out to discredit Nelson-Rees's years of crusading, as he had somehow imagined. It had all begun as a simple study of chromosomes in fresh tissue samples. Pathak wanted to find out if the defects observed in chromosomes of cultured cells resulted somehow from extended growth in a laboratory dish. Or did cancer cells taken fresh from a patient show abnormal chromosomes as well? Cailleau had been supplying him with fresh pleural effusions. And one day they simply stumbled across this unusual bunch of cells containing chromosomes similar to HeLa markers.

T.C. Hsu, the senior cytogeneticist in the lab, thought the finding raised an important point in the current climate of HeLaphobia. In most cases, said Hsu, perhaps 98 percent of them, a cell that has HeLa markers is probably HeLa. But these markers may not be unique to HeLaj they may be created by some general mishap of cancer. So there may be a few cases in which a cell carries HeLalike chromosomes and yet is something quite different.

"The findings so shocked Nelson-Rees, he thought it rocked the foundations of all his work," explained Hsu. "But it didn't really rock his foundations."

"We were simply urging caution when diagnosing cells as HeLa," added Pathak, "not disagreeing with all his results."

Still, in their published report, and in follow-up articles and letters to certain colleagues, Hsu, Pathak, and Cailleau repeatedly warned against relying on chromosomes alone, referring to Nelson-Rees by name and suggesting that he had foolishly done just that -- when he never had.

As for their need to create a collage, breaking with the traditional means of comparing chromosomes in cell lines, they explained that fresh tissue samples are always a mess, carrying all kinds of bodily by-products that are eventually filtered out of a long-term cell culture. So in a fresh pleural effusion, it is difficult to take clear photographs of all the marker chromosomes within a single cell.

"The composite was prepared in order to show an example that was clear-cut to the untrained eye," said Cailleau.

As Flandermeyer often pointed out, though, the identification of chromosomes entails a great deal of judgment. "There's an art to it, and if you're not careful you can incorporate your bias," he once said, thinking of the Denver affair. "You can make things conform to what you expect, especially if you pick around, choosing one chromosome from one cell and another from the next."

The presence of specific enzymes inside a cell is much less ambiguous, and therefore far more resistant, to an investigator's bias. On the other hand, the choice of which enzymes to look for and the order in which they are checked can create certain impressions.

Stephen O'Brien, an enzymologist at the National Cancer Institute and an ally of Nelson-Rees, was puzzled by the Anderson group's need to check fifteen enzymes in their breast cells before finding two that differed from HeLa's. Had he been doing the experiment, O'Brien said after reading the report, he would have analyzed perhaps six, and the two mismatches would have been among the first.

But according to Siciliano, the enzyme expert who collaborated with Hsu, Pathak, and Cailleau, the fifteen enzymes they checked were the standards, the ones they checked routinely. It just so happened the two that turned out to be different from HeLa's were tested last.

Like his colleagues, Siciliano insisted the report was not intended to discredit Nelson-Rees or to knock the foundation out from under his published findings. "I don't think there was any stuff in his published papers that's refutable," he said.

Why, then, the comment that Nelson-Rees's use of only three enzymes is overly simplistic?

"Most of the cells on his lists came from white donors," explained Siciliano, "so testing for only one enzyme, G6PD, was enough. But I was concerned for the future as more blacks donate tissue cultures, as they're doing in institutions like ours."

***

Motives, alas, are slippery fish. It is difficult for anyone to know what the Denver blowup was really all about. Anyone but Nelson-Rees, that is, to whom the truth was obvious.

"This group of people was trying to destroy years of careful work and promotion of the use of good cell lines, " he said after the incident. "It was just a case of this being put together to cause embarrassment to me.

"You know, I didn't 'impugn their reputations,' as some said of my rebuttal at Denver. I called them absolute shits. They were the ones who were unscientific and unethical, not me."
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:08 am

13: Even the Best of labs

One of the standard comebacks to Nelson-Rees's appeals was, "Ah, but HeLa threatens only the second-string team. Only the minor league players are sloppy enough to mix up their cells. For those of us who know what we're doing, this is all quite irrelevant."

In October 1978, Jonas Salk, developer of the Salk polio vaccine, founding director of the Salk Institute -- where some researchers practice philosophy as well as science -- medical giant Jonas Salk stepped forward as one large exception. He let it be known that HeLa cells had taken over one of his cell lines.

Salk had actually learned of this HeLa takeover years earlier, but hadn't spoken of it publicly until that October during a meeting of several hundred cell biologists and vaccine experts at the Lake Placid Lodge in New York. The eminent Dr. Salk was describing a series of experiments he had conducted on terminal cancer patients in the late 1950s. The idea was to try to switch the patients' immune systems into combat mode by challenging them with an injection of foreign cells. The foreign cells came from a line of monkey heart tissue, the same line that supplied the cultures in which he had grown polio viruses to develop his famous vaccine. Salk believed these monkey cells had certain antigens, certain biochemical identification tags, in common with cancer cells. He hoped that by activating the patients' immune systems against the monkey cells, he might trigger an attack against the cancer as well.

Strangely enough, Salk told his attentive audience, where he injected the cells, some of his patients developed tumors. The distinguished scientist-statesman flashed a few slides of human arms carrying pea- and almond-sized abcesses, and he said calmly that in retrospect he believed those cells may not have been his harmless monkey heart cells at all, but HeLa cells.

Well, a nervous murmur filled the auditorium and there were a couple of gasps as Salk's colleagues tried to envision him making the rounds with Henrietta Lacks hiding in his hypodermic, a determined Jonas Salk shooting up dying patients with an extra dose of overactive cancer cells.

Salk added quickly that the ugly little knots dissipated in no time, most of them in three weeks, never to return. Apparently he hadn't helped most of the patients with these injections -- although one man did undergo a remarkable remission and live another eighteen years -- but neither had he given them any more cancer than they already had. And that was the whole point of his talk: to report that even when injected directly into human beings, HeLa cells do not cause cancer.

A rather bizarre conclusion to present, perhaps, except that this was a conference about the making of vaccines. In particular it was about the use of cells in making vaccines. For the last few decades, there had been a running argument in this crowd over whether so-called continuous cell lines -- that is, established cultures that continue to reproduce themselves apparently without end -- are safe "substrates," safe soil, so to speak, in which to grow viruses to be used in vaccines. Or was there a chance the mysterious forces that give these cultures their immortality might also trigger runaway growth -- cancer, in effect -- in the recipients of vaccine prepared in those cells?

People like Salk argued there were adequate methods of separating a vaccine's active ingredients from the cells they were prepared in, so that it didn't matter what kind of cells were used. In fact Salk believed that even if vaccines weren't filtered at all, even if whole cancer cells were injected directly into a human test subject, they would be rejected by the body and do no harm. Of course no researcher would try that experiment. Not intentionally anyway. But twenty years earlier he had done it accidentally, Salk told the meeting, and only later did he realize it. He had injected HeLa cells into a few dozen patients, and it hadn't bothered them a bit.

Some of those who heard this remarkable talk later played it down, saying it came as no surprise. Virologists especially and people who were in on the early vaccine work said they had heard reports long before October 1978 that Salk's monkey heart cell showed some HeLalike characteristics. Nevertheless, that Salk had injected cervical tumor cells straight into human beings was disturbing to at least some of the conferees. There were also a few who thought Salk was awfully glib to conclude there was no danger in this simply because the lumps on patients' arms disappeared. Who knew what these HeLa cells had been up to inside each subject's body? The conference organizers apparently found this part of Salk's talk so unsettling they advised him to skip it in the written version to be submitted for publication. Sure enough, when the collection of reports was published the following year, Salk's paper made no mention of the inadvertent human experiments.

Nelson-Rees was one member of the audience who was openly shocked. To him, Jonas Salk had always been a renaissance scientist: brilliant, sophisticated, driven. That this "fantastic creature," as Nelson-Rees once described him, could be tripped up by HeLa tarnished the image considerably. On the other hand, here was a perfect demonstration of what Nelson-Rees had been preaching all along. Without eternal vigilance, HeLa can drop in on even the best of labs and the biggest of names.

Nelson-Rees, of course, had to be sure. Salk had said only that other scientists had reported his "monkey cells" to be HeLa; Salk had never checked them himself. So when it came time for questions, Nelson-Rees stood up and, as a few in the audience groaned and shook their heads, offered his services. If Salk still had samples of the twenty-year-old cultures, Nelson-Rees said, they could be clearly identified.

Salk graciously accepted Ne1son-Rees's very public offer.

The following week Nelson-Rees dropped in on Salk at his institute in La Jolla, a sprawling six-story concrete labyrinth built around a courtyard, all of it set upon high cliffs overlooking the Pacific. He was shown into Salk's office, a wood-paneled sanctum on the fifth floor. Gulls and hang-gliders rode the clouds just outside the window. Ne1son-Rees chatted briefly with Salk, picked up several samples of the cell line CH, cynomologous heart, and left for Oakland.

So, it does happen even to the first string, thought Nelson-Rees two months later, even to the titans of science. Salk's monkey cells showed no signs of monkey chromosomes. But, as Ne1son-Rees said in a letter to Salk, they did contain five of HeLa's "well-publicized marker chromosomes" as well as the type A G6PD enzyme.

Salk wrote back thanking Nelson-Rees for his work. He added that he looked forward to follow-up discussions.

***

But the follow-up discussions never took place. Salk's reaction, in fact, wasn't very different from many of HeLa's less illustrious victims. Not that he fought Nelson-Rees's conclusion in the scientific press or even argued with him personally. But deep down, he just couldn't buy it.

"It may well be the whole thing is due to contamination, but it bugs me every time I think about it in our own lab," he said a few years later. "Of course, no one is immune ... yet how it got there and how it could have happened is very mysterious to me."

This was not Relda Cailleau pounding a fist on the table. As he reminisced, he was relaxed, in control, speaking with the quiet intensity of a deep thinker.

"If it is due to contamination, then I ask myself, 'Why hasn't it been observed for other cells that are not HeLa? Why don't other cells crisscross? Why only HeLa?'"

These are the same questions Nelson-Rees had been asked a hundred times by scientists hoping to elude the stigma of a HeLa mix-up. So many researchers were under the impression that only HeLa preempted its fellow cell lines. Their argument went: "It just doesn't make sense that no other cell line would misbehave this way, and therefore it must not be happening."

But other cell lines do trespass on their neighbors. All the time. Besides his HeLa studies, Nelson-Rees had published reports listing dozens of nonHeLa mix-ups he had uncovered. Rat cells had overtaken a human breast culture, human cells had infiltrated a gibbon line, hamster cells were in where the marmosets were supposed to be, and dog cells were running wild among the minks. Sure, as an individual contaminator, HeLa had no equal. By no means, however, was it the only cell line that sneaked in where it didn't belong.

But Salk had another theory to offer: Isn't it possible that all cells in long-term culture take on certain common characteristics? We may think of these as HeLa's traits, but maybe they are the universal traits toward which all cells drift. If human beings emerged from monkeys and apes, picking up new traits as they evolved, why couldn't the same thing happen to cells in culture?

"Perhaps something fundamental occurs that expresses genes of evolutionary significance. This would not be a trivial observation," he said. He talked of silent genes gaining voice, "just an intuitive notion," you understand, "still in the form of an hypothesis." And yet maybe the apparent proliferation of HeLa cells is something far more significant. Maybe there's an alternative interpretation to these observations, something monumental, said Salk -- like Fleming's recognition of what the penicillin mold was doing to his staphylococcus bacteria!

This was nothing more than a warmed-over version of the argument Stan Gartler got when he unveiled the very first HeLa list in 1966. Wasn't it possible that some cells simply changed their G6PD enzyme from type B to A? Gartler and a Canadian researcher named Nellie Auersperg answered that question by growing one human cell line for three full years, monitoring three of its enzymes closely, and finding no such "evolution." No change even after they subjected the cultures to such triggers of evolutionary change as hormones, bacteria, viruses, and X rays. Cyril Stulberg and Ward Peterson at the Child Research Center in Detroit performed a related experiment a few years later, following the progress of a throat cancer line for two years with no sign of a change in its G6PD.

Similarly, in more than five years spent carefully examining human cell lines, Nelson-Rees never once saw a culture gradually develop HeLa marker chromosomes as if it were evolving into some universal HeLalike beast. When a HeLa takeover happened, it was always just as you'd expect in a cellular coup, quick and complete. Suddenly a culture had HeLa chromosomes, not just one but a whole family of markers, and they were accompanied by HeLa enzymes and other genetic markers that hadn't been there before; an across-the-board changeover, scarcely the kind of gradual metamorphosis that evolution would bring about.

Despite the evidence against it, this evolutionary theory of a "universal drift toward HeLa" had a loyal following.

"I don't doubt for a moment what Nelson-Rees observed in the monkey cells, but it's all a question of interpretation," said Salk. "I still don't want to throw out the notion that it may be due to evolutionary change.

"I'm trying to extract meaning out of these observations. Is this an example of evolution in the lab or is it contamination?"

A thoughtful pause.

"I'll entertain all the possible explanations until we know. I'm enough of a scientist to entertain both explanations."
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:09 am

14: The Little Dutch Boy

The wind blowing through Washington and into the National Cancer Institute suddenly shifted direction in the late 1970s. Part of it was a change in the political climate. Frank Rauscher resigned as director of the institute, and Jimmy Carter installed Arthur Upton as his replacement. Former director of the Brookhaven National Laboratory, Upton had long been interested in the health hazards of radiation. So it was only natural that under his guidance, the institute would become more concerned with the so-called environmental causes of cancer than it was under Nixon's man Rauscher, who was so thoroughly a virologist he even had a virus named after him.

But there was more going on than a simple changing of the chiefs.

By the late '70s, the quest for the viral cause of cancer simply didn't have much to show. There were a lot of amazing finds that stretched the bounds of basic biology and were leading to the genetic engineering revolution of the '80s. But there had been little progress in turning up the cause and the cure that everyone set out to find when the National Cancer Act was passed in 1971. It was now clear that cancer was a complex family of diseases with many possible causes. The unsung scientists who had been studying nonviral agents figured it was about time they were handed a larger hunk of the pie.

Meanwhile Americans were beginning to learn just how poisonous a land they lived in. In 1976, the institute published terrifying color-coded maps showing that specific cancers were native to certain parts of the country. The Northeast specialized in lung cancer, the Carolinas offered nose and throat cancer, and much of the Midwest and Texas featured leukemia. Suddenly that chemical stench along the northern end of the New Jersey Turnpike took on a horrible new significance, as did the haze over the refineries of the Gulf Coast and, later, those buried drums that turned up in the quiet community of Love Canal, New York. In the public mind, cancer was no longer the work of some mysterious microscopic bug, but something from the "environment," which is to say from the neighborhood -- something you could actually see, smell, and taste -- and that made it all the more terrifying.

In late 1978 the Carter administration pushed through a new law requiring federal researchers to test the carcinogenic powers of chemicals and low-level radiation. It called for lists to be published every year describing known or suspected cancer agents. And it mandated that the institute expand its study of cancer prevention, including such strategies as proper nutrition and limits on the spread of hazardous industrial materials and environmental pollutants. But because the institute's budget was to grow only slightly, something had to give to make way for the new priorities. And the most vulnerable part of the program was the viral cancer effort.

The resulting slowdown in viral cancer research could easily have threatened the Oakland lab, since cell lines had always been the raw material for viral work. Strangely enough, there were no signs of trouble during the late '70s. Oh, every now and then Jim Duff or some other visiting official would ask Nelson-Rees couldn't the Oakland operation be done on a smaller scale? Or couldn't they move it to Bethesda to be consolidated with some of the institute's other laboratories? But Nelson- Rees, convinced that the lab's prowess stemmed directly from its lavish endowment -- in good staff, in quality equipment, in plenty of time to be painstakingly careful -- would always argue loudly against it. So loudly, in fact, that even through a closed office door, staffers could hear him: "Just leave us alone so that we can do our jobs properly and well!" Nelson-Rees knew too that any operation ensconced in Bethesda would have none of the freedom that made Oakland so effective. He never raised this point, though. Mostly he ranted about how perfection requires resources, and they always dropped it.

No, the changing winds had not yet reached Oakland. In fact the turn of the decade was looking like boom time for Nelson-Rees's crusade. He and Stevenson finally convinced the editor of the Tissue Culture Association's journal, In Vitro, to require a full accounting of cell lines used in published studies. Now an author had to specify the supplier of the cells he studied and the various tests he used to confirm the presumed characteristics of the cells, including species, sex, race, and age of the donor. If he performed no such tests, he had to own up to that in the report. Nelson-Rees and Stevenson considered it a small but significant victory.

What's more, Stevenson left the institute's Frederick Cancer Center in 1979 to take over as director of the American Type Culture Collection. Ever since the early '60s, when he was at the institute trying to make the collection the nation's central cell repository, he felt it hadn't lived up to its first-class potential. As late as 1979 the place was still passing out some of the HeLa-contaminated cultures of the '60s under their original names and without clearly-worded warnings. The collection also had a bountiful supply of genuine cell lines that most researchers didn't seem to know about. As its new director, Stevenson aimed to clean the place up and then promote the hell out of it. With him at the collection and Nelson- Rees at Oakland, two of the world's most valuable biological stockpiles were being cared for by the two most devoted brothers of the Order of the Unspoiled Cell.

As it happened, the turn of the decade was an excellent season for HeLa hunting too. Nelson-Rees, Flandermeyer, and a recently hired technician named David Daniels uncovered HeLa cells masquerading as a liver culture in West Germany. They found HeLa lurking in three supposedly different cell lines from China, which they dubbed, "the gang of three." But of all the investigations that Nelson-Rees and the troops launched in this boom time, the case involving the cells of the little Dutch boy was the most impressive.

***

In October 1979 researchers at Penn State University published a report in Science supporting a controversial claim that tiny amounts of radiation can kill cells, cause genetic damage, and trigger cancer. They exposed human kidney cells known as T-1 to various levels of gamma radiation and found that some were killed even by extremely low doses. Nelson-Rees had never heard of the T-1 cell line, and he was puzzled by a statement in the report suggesting the culture was more than twenty years old. If, as the report implied, these cells were normal -- that is, noncancerous -- they would probably have gone through their allotted fifty to sixty doublings and died off long ago.

When he called Paul Todd, one of the authors, and identified himself, Todd said, "I had a feeling I would be hearing from you one of these days." Nelson-Rees could scarcely believe it when Todd explained that three years earlier he and his co-workers tried to submit their cell line to the American Type Culture Collection but were turned down because K. S. Lavappa, the collection's chromosome expert, found HeLa markers in it.

"Well then why did you go ahead and describe these as T-1 cells in Science?" asked Nelson-Rees. "Why didn't you state that they were HeLa?"

Because when they had tried to mention it in previous reports on the cell line, said Todd, journal reviewers asked that the explanation be deleted. Reviewers for Radiation Oncology, for instance, called the information "cell culture folklore" and said it was out of place in their journal. Besides, said Todd, the culture was only suspected of being contaminated; Lavappa's evidence was skimpy. To Todd, these T-1 cells didn't look anything like the HeLa cells he had seen.

Nelson-Rees wanted to judge for himself. He asked Todd to send him a sample of the kidney cells, then orchestrated an analysis so thorough it would be impossible to wave off with a claim of "too skimpy." He set Flandermeyer and Daniels to work on the chromosomes. He enlisted the help of Stephen O'Brien at the institute to test eight key enzymes instead of the usual two or three. And he got a colleague at the Scripps Research Institute in La Jolla to check nine HLA antigens.

While the testing was underway Nelson-Rees pieced together T-1's history. From a 1957 paper reporting the first culturing of the cells, he learned they had come from an eight-year-old Dutch boy who was operated on for kidney stones. A scientist named J. van der Veen put the tissue in a roller tube, where the cells grew for twelve days, then stalled. For months he transferred bits of the culture to new vessels, but none would grow. Then one of the bits suddenly blossomed. The cells spread over the entire tube in seven days and, carved up and placed into additional tubes, kept growing rapidly to become van der Veen's T-1 line.

The report read like such a classic HeLa takeover -- van der Veen mentioned he also had been working with HeLa at the time -- that Nelson-Rees had no doubt how the test results would turn out. By February 1980 Daniels and Flandermeyer had found four banded HeLa markers in some of the clearest, most convincing mugshots the lab had ever produced; they also reported the lack of any Y chromosomes, despite the fact that the donor was a boy. All nine HLA antigens tested in these "kidney cells" matched those of HeLa, as did all eight enzyme variants; the odds of these biochemical fingerprints being identical purely by chance were less than one in a million.

It was an airtight case against the Penn State culture. Having dug through the literature, though, Nelson-Rees realized that University Park, Pennsylvania, was only one of many stops the T-1 cell line had made. Over twenty years various strains of T-1 had become the favorites of radiation health researchers all around the world. To Nelson-Rees it looked as if a T-1 mafia had quietly gained control of the entire field. Much of what was known about the harm inflicted on human cells by radiation, and much of what influenced safety standards and guidelines to radiation exposure, was apparently based on experiments with T-1. He decided the case could not be closed until he had rounded up the entire gang.

The Penn State cells were ancestors of a sample of T-1 that Todd had obtained from researchers at Lawrence Berkeley Laboratory in California, who had also sent some to colleagues at the Los Alamos Scientific Laboratory in New Mexico. The Berkeley scientists got theirs from a Dutch researcher named C.W. Barendsen, who got his from van der Veen himself. Nelson-Rees retraced the trail, asking for samples from every lab along the way; he even got van der Veen to send a culture from a frozen stock of very early vintage. In August of 1980, Science published his reconstruction of the whole affair. It was an investigative tour de force. Five strains of T-1 cells from laboratories as far apart as Berkeley and Utrecht showed precisely the same incriminating characteristics of HeLa.

Science also ran a news analysis like the one that accompanied Nelson-Rees's first hit list. Titled "The Case of the Unmentioned Malignancy," the article made hay of the Penn State researchers' decision not to report the HeLa contamination of their cell line. Nelson-Rees tried to be tactful when asked for his comment. He noted the attempts by Todd's group to present the information in other, earlier reports. "I don't think that they swallowed the whistle," he was quoted as saying. "But they certainly didn't blow it." A couple of newspapers and magazines played the incident like a minor scandal.

The question of what HeLa's latest charade really meant for radiation research and existing standards, unfortunately, was never settled. Todd and associates claimed that even if T-1 were a HeLa culture, it wouldn't change their results. There was no evidence that a cancer cell would respond to radiation any differeJ1tly from a normal cell, they said. One biophysicist at the Argonne National Laboratory, however, had observed that tumor cells survive some types of radiation significantly better than normal cells. She wrote an angry letter to Science complaining about the Penn State study. Bob Stevenson joined the fray too, writing to Todd's group that it was "pretty shoddy" to use HeLa-contaminated cells in an experiment without revealing it. He was so angry, in fact, he also wrote to the institute's grant officers who were funding the Penn State team to make sure they knew about it.

All in all, it was a highly visible demonstration of the Oakland team's powers of investigation and prosecution. In celebration, Daniels and a few of the other lab workers presented Nelson-Rees with a small gardening tool labeled "official muck rake."

Just about that time, an ominous bunch of letters went out from the institute's biological carcinogenesis branch, the direct descendant of the old viral cancer office. In light of "possible budget reductions," the letters said, the institute was polling people who recently had obtained cells from Nelson-Rees or had asked his lab to identify unknown cultures. How heavily did these people depend on the cells and the expertise in Oakland, the letters asked, and what impact would it have on their work if they had to pay a commercial laboratory for these services, which the institute had been supplying to them for free?

Nelson-Rees got a letter too. It directed him to describe the lab's activities and explain why they were necessary.

Were they sincere? Did they actually have to be told why it was necessary to safeguard the most widely used experimental material in cancer research? Could it be, Nelson-Rees wondered, that they were unacquainted with one of the basic principles of the scientific method: to know what the hell one is working with? And was Walter Ne1son-Rees, the keeper of the cells, actually being ordered to account for himself? It was an outrage.

He considered sending out his own letter to researchers asking how heavily they depended on the institute's biological carcinogenesis branch and what would be the impact of phasing out a few of the bureaucrats there. But then it occurred to him that maybe this wasn't simply another argument he would win by shouting down Jim Duff. Maybe this time they were serious.

In fact, for the first time since America declared war on cancer, the institute's total budget was about to shrink: from $958 million in 1980 to $947 million for the upcoming year. The funding for viral research and related activities, already on the slide, would be cut back even more.

The shifting winds from Washington had finally reached Oakland. Shortly before Christmas 1980, the bureaucrats notified Nelson-Rees that his $600,000 annual budget would be trimmed by 20 percent. He would have to fire six of his fifteen people.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

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15: Battle Fatigue

In February of 1981, Bob Flandermeyer attended a conference in San Francisco on genetic engineering, which, unlike most scientific conferences Flandermeyer had been to, was crawling with corporate researchers. The manipulation of genes was becoming big business.

Flandermeyer went to a talk about interferon, a natural protein that showed some ability to fight off viruses and control the spread of certain cancers. Recently featured on the cover of Time, interferon was now being synthesized by many of the new firms, whose directors hoped to cash in on its wonder-drug potential. Flandermeyer heard a fellow from Genentech, one of the giants in the field, describe how they tested the potency of their interferon. They treated several cultures of cells with different doses of the chemical, then rated how well those cells withstood assaults by hostile viruses. The cell line routinely used for the test, said the man from Genentech, was a culture of normal human amnion tissue called WISH.

Flandermeyer thought WISH sounded familiar, but before saying anything he telephoned the lab.

"WISH, of course!" bellowed Nelson-Rees. "Wistar Institute Susan Hayflick. That was the amniotic sac in which Leonard Hayflick's daughter was delivered. He made a big joke about it when Stan Gartler claimed it was HeLa in 1966."

Flandermeyer said he'd better go tell the genetic engineer.

Over the next few weeks, Nelson-Rees thought a lot about that simple phone conversation. It occurred to him that the current boom in bioengineering was very different from the biomedical boom in the '70s. The motivation then was "The War." The glorious dream of conquering cancer had pumped billions of dollars into research and attracted hundreds of eager recruits. Now the motivation was profit. These outfits were businesses, and the pressure to be the first with a new product stemmed from commercial competition. Furthermore, proprietary information and corporate secrets were the standard rules by which these new people played. Businessmen don't openly share the details of their manufacturing processes, even if their factories are living cells and bacteria, their foremen molecular biologists.

Not that there was anything wrong with Cenentech's using HeLa cells to test interferon. Since the cells were serving strictly as targets for viruses, the potency ratings were probably valid as long as the company always used the same cell line. What bothered Nelson- Rees was that the people at Cenentech were calling these cells human amnion. They didn't know. They might never have known if Bob Flandermeyer hadn't wandered in and told them. And if they didn't know in this case, they might not know in cases where it really matters.

After all, these researchers were playing around with genes, snipping a few out of one cell and splicing them in among those of another. Well, scientists knew enough to say that genes control everything, but they had barely a clue about which of the thousands of individual genes directed this function or determined that characteristic. Cancer researchers, for instance, had just lately found out about the oncogenes, a mysterious bunch of genetic elements that usually sit dormant upon the human chromosome but that may under certain conditions in certain cells "turn on" and trigger cancer. Which only meant to Nelson-Rees that as these bioengineers are cutting and pasting up genes to create hormones and drugs and other things intended to go into human bodies, they'd better be damned sure they know what cells they're finagling.

That one short phone call from Flandermeyer made Nelson-Rees realize this was probably one of the worst times to abandon the crusade, and yet that's exactly what he was considering doing. It had been frustrating for him these last few months: trying to run the cell culture lab at less than full strength and at the same time keep the crusade going. Everyone was putting in extra hours. Still he knew the cutbacks had only begun, and any added load would be too much to handle if they were to keep up the quality of work that the lab -- that he -- had always been known for. David Daniels said the place was starting to take on the look of "a ma-and-pa operation." That phrase stuck with Nelson-Rees, grating on his perfectionism like a burr in his boot.

The funding cutback was really the last of a series of frustrations. Although he rarely showed it and never discussed it even with close friends, Nelson-Rees was weary of the battle. He was in a business that generated enemies, not allies. Even the Tissue Culture Association, the scientific group that ought to have been cheerleading him on, only tolerated him after the blow-up in Denver several years earlier. The latest order from the institute to defend the lab's existence in spite of what he felt was a glorious record reminded him once again that not everyone thought his contribution was so vital.

"I don't think Walter felt anyone fully appreciated the value of the work he had done," Jim Duff once observed. "You know, they don't award Nobel Prizes for finding out that things are wrong."

So in March Nelson-Rees called the staff together and gave a subdued little speech about the cutbacks, present and future, saying in short that you can't fight a crusade without funding. Difficult as it would be, he said, he would be quitting toward the end of the year. Not just quitting the lab and moving on, but leaving research entirely. He planned to devote his time to the small art gallery he and a friend had been operating out of his home the past few years.

Quitting, just like that.

***

Nelson-Rees figured a scaled-down cell culture lab would continue at Oakland. But after his resignation the bureaucrats announced that the facility would be shut down completely by the end of the next year. It looked as if he had played right into their hands. They said they would arrange to have the 2,000 cell cultures transferred to the freezers of the American Type Culture Collection, although they could provide no funds for the maintenance and distribution of the cells. As for the Oakland lab's service of checking the identity of cultures, they weren't setting up any substitute for that. Ward Peterson in Detroit could do the enzyme testing he had always done as well as some of the simpler chromosomal checks. But researchers would have to pay for the analysis.

"I'll have to think very carefully now before sending some culture out to be checked," said one candid scientist when asked about the loss of the Oakland lab. "When it was all free, we sent things out all the time. It was a tremendous help. But now, at two or three hundred dollars a shot .... "

Even more than the loss of the service, though, some mourned the loss of the crusader himself. Peterson wrote to Nelson-Rees, "As the most articulate amongst a small group that has promulgated the rules, your role as the point man will greatly be missed." That was the real worry of the few cancer researchers and cell biologists who had appreciated Nelson-Rees. Who was to carry on? Peterson was concerned about cell line screw-ups, but he was no zealot. Stevenson would continue his missionary work, of course, in a tactful and politically sensitive way. But challenging colleagues at public meetings, naming names in journals, making an occasional scene -- that just wasn't their style.

There were a few protests to the institute. One scientist called Nelson-Rees "a national resource," and estimated that by spotting suspicious cell lines he had probably prevented the waste of tens of millions, if not hundreds of millions of research dollars. The institute was unimpressed. The feeling, at least among certain officials, was that these worries were held over from the old days, the unenlightened days when cell culture needed to be actively policed.

"Nelson-Rees may have been exactly the right person for that time," explained one bureaucrat. "A more subtle and conciliatory style might not have spread knowledge of the extent of HeLa contamination throughout the world back then. Today, however, I would hope that there is sufficient awareness and sufficient expertise that any of these potential things would be avoided or caught quickly."

With Nelson-Rees off the scene, that was about all they could do: hope that nothing went wrong.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:15 am

16: Legacy

In a cramped and cluttered office at Montefiore Hospital in the Bronx, there stands a kind of monument to Nelson-Rees and his crusade. Tacked onto the door frame is a slightly yellowed copy of his third and final hit list, published in April 1981, just a few weeks after he announced he was quitting.

This is the office of Fritz Herz, chief of the tissue culture section in the hospital's pathology department. Five times a year, Herz holds a seminar in the art of culturing cells. He starts with a demonstration of the basics, reminding his students never to feed different cells from the same pipette -- "Look," he tells them, "I'm throwing the pipette away" -- and ends with the story about Henrietta Lacks, Walter Nelson-Rees, and the list.

Things are not always what they seem, Herz explains, pointing to the three full pages that feature such oldies as WISH and Relda Cailleau's MAC-21 plus twenty-two new HeLa-contaminated cultures uncovered by Nelson-Rees, Flandermeyer, and Daniels in the last couple of years they worked together. Ninety cell lines in all. Bob Stevenson guessed that was about a third of the more popular cell lines used in cancer and related research: one out of every three cell lines was an impostor.

"It's very important that you know what your cells are, that you know they are not HeLa," Herz tells the residents, the neuropathologists, and the other young scientists who come through his lab. "If you're sloppy and you don't care and you're not committed to perfection, you'll soon find your own work printed on a list like this."

That was how Nelson-Rees left his mark. A few loyal fans, a couple of disciples who preached in their own small parishes. He had untangled much of the morass created by Henrietta Lacks's runaway cells, but once he left his post, there was no guarantee that things would stay untangled. The bureaucrats were hoping that the collective consciousness of medical research had been propped up to where it no longer needed to be nagged about sloppy technique and wasteful mix-ups. But if ever there was a persuasive argument against that kind of thinking, this list was it. Just because Nelson-Rees had got out of the business, its ninety entries seemed to say, that didn't mean the ghost of Henrietta Lacks was retiring.

Which proved to be absolutely correct.

Microbiological Associates, for example, the company that created the He La-contaminated culture called MA160 thirteen years before the publication of this last list, had changed its name to M.A. Bioproducts but made no change in its catalogue description of MA160. Although Nelson-Rees and half-a-dozen independent scientists had concluded by the mid-'70s that the culture was nothing but cervical cancer cells from a black woman, the company continued selling it through 1981 as "prostate, benign, human adult."

In the fall of that same year, as Nelson-Rees was cleaning out his file cabinets in Oakland, a new study on prostate cancer was published. It was based on experiments with MA160 and its identical, HeLa-contaminated twin EB33, the culture Nelson-Rees had condemned in 1976. The following year, there appeared another study based in part on these two adulterated cultures.

In January 1983 a team of Dutch, Finnish, and American researchers announced their discovery that the U cell line, presumed to be a culture of amnion cells taken from a Dutch woman in 1957, was actually HeLa.

And in the spring, Nelson-Rees spoke with a young San Diego researcher named Mark Bogart, who had recently checked thirty different cell lines from various scientists and found that half were not what these scientists had thought. Bogart's methods were unsophisticated compared to the techniques Nelson-Rees had used. Like the early tissue culturists in the 1950s, he could only tell species apart. He knew that two mouse lines had been jumped by a culture of human cells, for example, but he couldn't say which human culture that was. He knew that ten of the supposedly human cultures he analyzed were indeed human, but he had no idea whether they held the right human cells or -- who knows? -- perhaps the cells of a certain lady from Baltimore.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

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17: Epilogue

Walter Nelson-Rees is no longer chasing Henrietta Lacks, but he keeps in touch with a few colleagues and peruses the journals regularly enough to know that she still haunts the biomedical labs. And sitting in his home in Oakland, next to a wall filled with impressionist paintings, he is happy to deliver a private sermon when asked why she endures.

There's more to the problem than a tenacious and hardy cell culture, he says. HeLa cells persist because they have always been helped along by a certain human element in science, an element connected to emotions, egos, a reluctance to admit mistakes, and many of the other things he used to lecture about.

"It's all human -- an unwillingness to throwaway hours and hours of what was thought to be good research, worries about jeopardizing another grant that's being applied for, the hurrying to come out with a paper first. And it isn't limited to biology and cancer research. Scientists in many endeavors all make mistakes, and they all have the same problems."

Wade Parks, the virologist who in 1972 exposed the famous Russian cells as worthless carriers of a monkey virus, once said something similar. In every field, he observed, this human factor encourages "HeLas."

"A 'HeLa,'" Parks explained, "is a scientific claim that sucks people into a line of work for a while, a line that is later refuted or shown to be a waste of time. It's a type of error in science that occurs fairly often. And it will continue to exist."  
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:17 am

Acknowledgments

The writing of this book depended upon the help of scores of people, each of whom spent hours with me, recalling events, emotions, even conversations that took place as long ago as twenty years. For their willingness to relive those times and to be candid about them, I am very grateful. My thanks also to Robert Stevenson of the American Type Culture Collection for allowing me access to the files of the former Berkeley Cell Culture Laboratory, of which he is now the custodian. I am grateful, too, to the people at Science 85 magazine for allowing me a brief leave of absence to get started on this book and for putting up with me after I returned to work and continued writing it in my "spare time." Many thanks to Jack Codler, who cajoled me into undertaking this project in the first place, and to Susan Zeckendorf, who shared her knowledge of publishing and her great enthusiasm.

My special gratitude goes to two people who served as unofficial editors and tireless cheerleaders: Susan West and Eric Schrier. Their detailed critiques and suggestions greatly improved every draft chapter. And without their genuine support, this book might never have been completed. Finally, to Susan, who while editing and cheerleading also offered love and patience, my greatest thanks.
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Re: A Conspiracy of Cells: One Woman's Immortal Legacy and t

Postby admin » Mon Mar 07, 2016 2:27 am

Sources

The following references offer some historical perspective to the events described, as well as greater technical depth for those who are interested. It is by no means comprehensive.

Chapter 1. Special Delivery

"Isolation and Culture of a Virus from Human Cancer Tissue." Science News, 10 July 1971, p. 21.

"Race for Human Cancer Virus: Odds against Houston Team Lengthen." Science, 24 September 1971, p. 1220.
"USA and USSR Communication in Cancer Research," by Joseph F. Saunders, Biosciences Communications, 2, volume 2, (1976) p. 98.

Chapter 2. The Seed That Took

"Some Aspects of the Constitution and Behavior of Normal and Malignant Cells Maintained in Continuous Culture," by George O. Gey, The Harvey Lectures: Series L (1954-1955), Academic Press Inc., New York, N.Y., p. 154.

"George Otto Gey: The HeLa Cell and a Reappraisal of its Origin," by Howard Jones Jr., Victor A. Mckusick, Peter S. Harper, and Kuang-Dong Wuu, Obstetrics and Gynecology, December 1971, p. 945.

"History of Tissue Culture at Johns Hopkins," by Frederick B. Bang, Bulletin of the History of Medicine, volume 51, 1977, p. 516.

Chapter 3. HeLagram

"Studies on the Propagation in Vitro of Poliomyelitus Viruses," by William F. Scherer, Jerome Syverton, and George O. Gey, Journal of Experimental Medicine, volume 97, 1953, p. 695.

"Continuous subcultivation of epithelial-like cells from normal human tissues," by R.S. Chang, Proceedings of the Society for Experimental Biology and Medicine, volume 87, 1954, p. 440.

"The origin of altered cell lines from mouse, monkey, and man, as indicated by chromosome and transplantation studies," by K.H. Rothfels, A.A. Axelrad, L. Siminovitch, E.A. McCulloch, and R.C. Parker, Proceedings of the Third Canadian Cancer Conference, 1959, p. 189.

"The establishment of a line (WISH) of human amnion cells in continuous cultivation," by Leonard Hayflick, Experimental Cell Research, volume 23, 1961, p. 14.

"Results of tests for the species of origin of cell lines by means of the mixed agglutination reaction," by D. Franks, B.W. Gurner, R.R.A. Coombs, and R. Stevenson, Experimental Cell Research, volume 28, 1962, p. 608.

"Collection, Preservation, Characterization, and Distribution of Cell Cultures," by Robert E. Stevenson, Proceedings of the Symposium on the Characterization and Uses of Human Diploid Cell Strains, Opatija, Yugoslavia, 1963, p. 417.

"Genetic Markers as Tracers in Cell Culture," by Stanley M. Garder, National Cancer Institute Monograph No. 26: Second Decennial Review Conference on Cell Tissue and Organ Culture, 1967, p. 167.

"Apparent HeLa Cell Contamination of Human Heteroploid Cell Lines," by Stanley M. Garder, Nature, February 24, 1968, p. 750.

"Cell Culture Collection Committee in the United States," by Robert E. Stevenson, Cancer Cells in Culture, edited by H. Katsuta, University of Tokyo Press, 1968, p. 385.

"Extrinsic Cell Contamination of Tissue Cultures," by Cyril S. Stulberg, Contamination in Tissue Culture, edited by Jorgen Fogh, Academic Press, New York and London, 1973, p. 1.

Chapter 4. Out of Thin Air

"Oncogenesis in Vitro," by Leonard Hayflick, National Cancer Institute Monograph No. 26: Second Decennial Review Conference on Cell Tissue and Organ Culture, 1967, p. 355.

"Nutritional Needs of Mammalian Cells in Tissue Culture," by Harry Eagle, Science, September 16, 1955, p. 501.

"Common Antigens in Tissue Culture Cell Lines," by Lewis L. Coriell, Milton G. Tall, and Helen Gaskill, Science, July 25, 1958, p. 198.

"Detection and Elimination of Contaminating Organisms," by L. Coriell, National Cancer Institute Monograph No. 7, April 1962, p. 33.

"Glucose-6-phosphate Dehydrogenase Isoenzymes in Human Cell Cultures Determined by Sucrose-Agar Gel and Cellulose Acetate Zymograms," by w.O. Peterson Jr., C.S. Stulberg, N.K. Swanborg, and A.R. Robinson, Proceedings of the Society of Experimental Biology and Medicine, volume 128, 1968, p. 772.

"The Animal Cell Culture Collection," by C.S. Stulberg, L.L. Coriell, A.J. Kniazeff, and J.E. Shannon, In Vitro, volume 5, 1970, p. 1.

"Oncornavirus-like particles in He La Cells," by H. Bauer, J.H. Daams, KF. Watson, K Molling, H. Gelderblom, and W. Schafer, International Tournal of Cancer, volume 13, 1974, p.254.

"A Quest for the Mechanism of 'Spontaneous' Malignant Transformation in Culture with Associated Advances in Culture Technology," by Katherine K Sanford and Virginia J. Evans, Journal of the National Cancer Institute, June 1982, p. 895.

Chapter 5. In The Purple Palace

"Isolation of Oncornaviruses from Continuous Human Cell Cultures," by Victor M. Zhdanov, Valentine D. Soloviev, Tagir A. Beketemirov, Konstantin V. Ilyin, Albert F. Bykovsky, Nikolai P. Mazurenko, Iosif S. Irlin, and Felix I. Yershov, Intervirology, volume 1, 1973, p. 19.

"Mason-Pfizer virus characterization: A similar virus in a human amniotic cell line," by W.P. Parks, R.V. Gilden, A.F. Bykovsky et al., JournaI of Virology, volume 12, 1973, p. 1540.

"HeLa-Like Marker Chromosomes and Type-A Variant Glucose-6-phosphate Dehydrogenase Isoenzyme in Human Cell Cultures Producing Mason-Pfizer Monkey Virus-Like Particles," by W.A. Nelson-Rees, V.M. Zhdanov, P.K. Hawthorne, and R.R. Flandermeyer, TournaI of the National Cancer Institute, September 1974, p. 751.

Chapter 7. Mug Shots

"Quinacrine fluorescent karyotypes of human diploid and heteroploid lines," by O.J. Miller, D.A. Miller, P.W. Allderdice, V.G. Dev, and M.S. Grewal, Cytogenetics, volume 10, 1971, p. 338.

"Isolation of a Continuous Epithelioid Cell Line, HBT-3, from a Human Breast Carcinoma," by Robert H. Bassin, Ernest J. Plata, Brenda I. Gerwin, Carl F. Mattern, Daniel K. Haapala, and Elizabeth W. Chu, Proceedings of the Society for Experimental Biology and Medicine, November 1972, p. 673.

"An Established Cell Line (HBT-39) From Human Breast Carcinoma," by Ernest J. Plata, Tadao Aoki, Diane D. Robertson, Elizabeth W. Chu, and Brenda I. Gerwin, Journal of the National Cancer Institute, April 1973, p. 849.

"Banded Marker Chromosomes as Indicators of Intraspecies Cellular Contamination," by Walter A. Nelson-Rees, Robert R. Flandermeyer, and Paula K. Hawthorne, Science, June 7, 1974, p. 1093.

"HeLa Cells: Contaminating Cultures around the World." by Barbara J. Culliton, Science, June 7, 1974, p. 1059.

"Human Breast Tumor Cell Lines: Identity Evaluation by Ultrastructure," by Gertrude C. Buehring and Adeline J. Hackett, Journal of the National Cancer Institute, September 1974, p.621.

Chapter 8. Spreading the Word

"The establishment of a Cell Strain (MAC-21) from a Mucoid Adenocarcinoma of the Human Lung," by Relda Cailleau, Cancer Research, July 1960, p. 837.

"Two Cell Lines (SH-2 and SH-3) Derived from Human Breast Cancer," by G. Seman, S.J. Hunter, and 1. Dmochowski, Proceedings of the American Association for Cancer Research, volume 16, March 1975, p. 59.

Chapter 9. Damage Report

"Detection and Isolation of a New DNA Polymerase from Human Breast Tumor Cell Line HBT-3 by (dT) 12-18-Cellulose Chromatography," by Brenda 1. Gerwin and Robert H. Bassin, Proceedings of the National Academy of Sciences, August 1973, p. 2453.

"Present Status of MA160 Cell Line, Prostatic Epithelieum or HeLa Cells?" by Mukta M. Webber, Paul K. Horan, and Thomas R. Bouldin, Investigative Urology, volume 14, 1977, p. 335.

Chapter 10. Provenance

"Spontaneous in vitro Neoplastic Transformation of Adult Human Prostatic Epithelium," by Elwin E. Fraley, Sidney Ecker, and Monroe M. Vincent, Science, October 30, 1970, p. 541.

"Human Prostatic Carcinoma in Cell Culture: Preliminary Report on the Development and Characterization of an Epithelial Cell Line (EB33)," by K. Okada and F.H. Schroeder, Urological Research, volume 2, 1974, p. 111.

"Human Breast Cancer Cells in Continuous Cultivation Used to Determine Chemotherapy Sensitivity of the Patient from Whom They Are Derived," by Roland A. Pattillo and A.C.F. Ruckert, Proceedings of the American Association for Cancer Research, volume 16, March 1975, p. 145.

"HeLa Cultures Defined," by Walter A. Nelson-Rees and Robert R. Flandermeyer, Science, January 9, 1976, p. 96.

"Examination of ATCC stocks for HeLa marker chromosomes in human cell lines," by K.S. Lavappa, M.L. Macy, and J.E. Shannon, Nature, January 22, 1976, p. 211.

"HeLa takes over," by Sandy Grimwade, Nature, January 22, 1976, p. 172.

"Cell Cultures: Confused and Contaminated," Science News, volume 109, 1976, p. 36.

"Characterization of an established cell line (SH 3) derived from pleural effusion of patient with breast cancer," by G. Seman, S.J. Hunter, R.C Miller, and L. Dmochowski, Cancer, volume 37, 1976, p. 1814.

Chapter 11. Another Run-in With Relda

"Human Lung Organ-Specific Antigens on Normal Lung, Lung Tumors, and a Lung Tumor Cell Line," by Richard Akeson, Journal of the National Cancer Institute, April 1977, p. 863.

"Lung Organ-Specific Antigens on Cells with HeLa Marker Chromosomes," letters by Walter A. Nelson-Rees, R.A. Akeson, and Relda Cailleau, Journal of the National Cancer Institute, June 1978, p. 1205.

Chapter 12. Showdown

"HeLa cells and their possible contamination of other cell lines: Karyotype studies," by W.K. Heneen, Hereditas, volume 82, 1976, p. 217.

"Foreword," by T.C Hsu, Mammalian Chromosome Newsletter, November 4, 1978, p. 103.

"A Human Breast Adenocarcinoma With Chromosome and Isoenzyme Markers Similar to Those of the HeLa Line," by Sen Pathak, Michael J. Siciliano, Relda Cailleau, Charles L. Wiseman, and T.C Hsu, Journal of the National Cancer Institute, February 1979, p. 263.

"A Molecular Approach to the Identification and Individualization of Human and Animal Cells in Culture: Isozyme and Allozyme Genetic Signatures," by Stephen J. O'Brien, John E. Shannon, and Mitchell H. Gail, In Vitro, volume 16, 1980, p. 119.

Chapter 13. Even the Best of Labs

"Some Characteristics of a Continuously Propagating Cell Derived from Monkey Heart Tissue," by Jonas E. Salk and Elsie N. Ward, Science, December 27, 1957, p. 1338.

"Immunological Paradoxes: Theoretical Considerations in the Rejection or Retention of Grafts, Tumors, and Normal Tissue," by Jonas Salk, Annals of the New York Academy of Sciences, October 14, 1969, p. 365.

"Isozyme stability in human heteroploid cell lines," by N. Auersperg and S.M. Gartler, Experimental Cell Research, volume 61, 1970, p. 465.

"A Permanent Heteroploid Human Cell Line with Type B Glucose-phosphate Dehydrogenase," by W.O. Peterson Jr., C.S. Stulberg, and W.F. Simpson, Proceedings of the Society for Experimental Biology and Medicine, April 1971, p. 1187.

"Inter- and Intraspecies Contamination of Human Breast Tumor Cell Lines HBC and BrCa5 and Other Cell Cultures, by Walter A. Nelson-Rees and Robert R. Flandermeyer, Science, March 25, 1977, p. 1343.

"The Identification and Monitoring of Cell Line Specificity," by Walter A. Nelson-Rees, The Origin and Natural History of Cell Lines, edited by Claudio Barigozzi, Alan R. Liss, Inc., New York, 1978, p. 25.

"The Spector of Malignancy and Criteria for Cell Lines as Substrates for Vaccines," by Jonas Salk, Cell Substrates and Their Use in the Production of Vaccines and Other Biologicals, edited by John C. Petricciani, Hope E. Hopps, and Paul Chapple, Plenum Press, New York and London, 1979, p. 107.

"Characteristics of HeLa strains: Permanent vs. variable features," by W.A. Nelson-Rees, L. Hunter, G.J. Darlington, and S.J. O'Brien, Cytogenetics and Cell Genetics, volume 27, 1980, p. 216.

Chapter 14. The Little Dutch Boy

Atlas of Cancer Mortality for U.S. Counties: 1950 - 1969, by Thomas J. Mason, Frank W. McKay, Robert Hoover, William J. Blot, and Joseph F. Fraumeni Jr., U.S. Department of Health, Education, and Welfare Publication No. (NIH) 75-780.

"Establishment of Two Human Cell Strains from Kidney and Reticulosarcoma of Lung," by J. van der Veen, 1. Bots, and A. Mes, Archiv fur die Gesamte Virusforschung, volume 8, 1958, p. 230.

"Comparison of the Effects of Various Cyclotron-Produced Fast Neutrons on the Reproductive Capacity of Cultured Human Kidney (T-1) Cells," by Paul Todd, Joseph Geraci, Paul S. Furcinitti, Randall M. Rossi, Fuminori Mikage, Richard B. Theus, and Carter B. Schroy, International Journal of Radiation Oncology, Biology, Physics, volume 4, 1978, p. 1015.

"The Effects of Caffeine on the Expression of Potentially Lethal and Sublethal Damage in Gamma-Irradiated Cultured Mammalian Cell," by Carter B. Schroy and Paul Todd, Radiation Research, volume 78, 1979, p. 312.

"Inactivation of Human Kidney Cells by High-Energy Monoenergetic Heavy-Ion Beams," by Eleanor A. Blakely, Cornelius A. Tobias, Tracy C.H. Yang, Karen e. Smith, and John T. Lyman, Radiation Research, volume 80, 1979, p. 122.

"Gamma Rays: Further Evidence for Lack of a Threshold Dose for Lethality to Human Cells," by Paul S. Furcinitti and Paul Todd, Science, October 26, 1979, p. 475.

"T-1 Cells Are HeLa and Not of Normal Human Kidney Origin," by Walter A. Nelson-Rees, Robert R. Flandermeyer, and David W. Daniels, Science, August 8, 1980, p. 719.

"Hendrik or Henrietta?" The Economist, November 15, 1980, p. 104.

"The Case of the Unmentioned Malignancy," by William J. Broad, Science, December 12, 1980, p. 1229.

Chapter 16. legacy

"Cross-Contamination of Cells in Culture," by W.A. Nelson-Rees, D.W. Daniels, and R.R. Flandermeyer, Science, April 24, 1981, p. 446.

"The Surface Character of Separated Prostatic Cells and Cultured Fibroblasts of Prostatic Tissue as Determined by Concanavalin-A Hemadsorption," by K. Oishi, J.C. Romijn, and F.H. Schroeder, The Prostate, volume 2, 1981, p. 11.

"Monoclonal Antibodies to Human Prostate and Bladder Tumor-associated Antigens, II by James J. Starling, Susan M. Sieg, Mary L. Beckett, Paul F. Schellhammer, Leopoldo E. Ladaga, and George L. Wright Jr., Cancer Research, August 1982/ p. 3084.

"U cells contain contaminants, II by William C. Wright, Sara Kaffe, Christian F. Holinka, Kari Cantell, Jacoba G. Kapsenberg, and Kurt Hirschhorn, Nature, January 27, 1983, p. 279.
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