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.