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Germ warfare: the battle for the key to modern vaccines

In the late 1960s the scientist behind the worlds most successful antiviral vaccines went into battle against his employer and the US government for custody of the cells that he called his children

On 9 October 1964, a baby girl was born at Philadelphia general hospital. She arrived early, when her mother was about 32 weeks pregnant. The baby weighed 3.2lb and was noted to be blue, floppy and not breathing. The only sign of life was her slow heartbeat. Nonetheless, she clung on, and her 17-year-old mother named her.

One month later, the baby was still in the hospital, and a doctor listening with a stethoscope heard a harsh heart murmur. A chest X-ray showed that she had a massively enlarged heart because a hole in the organ was preventing it from pumping blood efficiently. It also emerged that the baby had cataracts blinding both eyes. Later, other signs indicated that she was profoundly deaf.

The baby also suffered from recurring respiratory infections and had trouble gaining weight. A psychologist who assessed her in July 1965 judged the nine-month-old to be the size of a two- or three-month-old infant and at about that stage of development, too. She needed heart surgery if she was going to survive. Just before her first birthday, surgeons made an incision in her chest wall and repaired her heart. After the operation, she remained in hospital. The chronic respiratory infections continued. The baby was 16 months old and weighed just 11lb when she died of pneumonia on 18 February 1966.

The young mother had told the doctors that when she was one month pregnant, she had contracted German measles, also known as rubella.

The early 1960s marked a coming of age for the study of viruses such as the one that causes rubella tiny infectious agents that invade cells and hijack their machinery in order to reproduce themselves. Biologists, with new tools in hand, were racing to capture viruses in throat swabs or urine or even snippets of organs from infected people and to grow them in lab dishes. Isolating a virus in the lab made it possible to make a vaccine against it. And making antiviral vaccines promised huge inroads against common childhood diseases such as measles, mumps and rubella, along with less common killers including hepatitis. The principle of vaccination is simple: if a person is injected with, or swallows, a tiny amount of a virus either a killed virus or a weakened live virus that person will develop antibodies against the virus. Then, if he or she is exposed in the future to the naturally occurring, disease-causing form of the virus, those antibodies will attack the invader and prevent it from causing disease.

But if the concept is simple, making effective vaccines is anything but. In the early 1960s, that reality was all too evident. In 1942, as many as 330,000 US servicemen were exposed to the hepatitis B virus in a yellow fever vaccine that was contaminated with blood plasma from infected donors (the plasma was used to stabilise the vaccine). Around 50,000 of the vaccinated servicemen contracted the liver disease and up to 150 died.

In 1955, a California-based company named Cutter Laboratories made a polio vaccine with the live, disease-causing virus in it. As a result, 192 people were paralysed many of them children and 10 died. Every senior US government employee involved in the Cutter incident lost his or her job, right up to the director of the National Institutes of Health (NIH) and the secretary for health, education and welfare.

Then, in the summer of 1961, Americans learned that cells used to manufacture the widely used Salk polio vaccine, harvested from monkey kidneys, harboured a virus named SV40. Tens of millions of American children had already received contaminated injections, and while the jury was still out on the tainted vaccines long-term health consequences, the risks were of great concern to regulators in the US and further afield.

It was against this backdrop that, on a drizzly June morning in 1962, a 34-year-old scientist named Leonard Hayflick went to work in his lab at the Wistar Institute of Anatomy and Biology an elegant 1890s brownstone tucked in the heart of the University of Pennsylvanias campus.

A serious, slight man with close-cropped dark hair, Hayflick was a product of working-class Philadelphia and hungry to make his name. He was in love with biology and had come to believe that he was extremely smart a fact that was far from appreciated. Hayflicks boss, the polio-vaccine pioneer Hilary Koprowski, saw him as a mere technician, hired to serve up bottles of lab-grown cells to the institutes scientists.

The ambitious Hayflick was undeterred. That day, he planned to launch a group of human cells that would revolutionise vaccine-making. He was convinced that, compared with monkey cells, which were often laden with viruses, human cells would serve as cleaner, safer vehicles for producing antiviral vaccines.

Several days earlier, a woman living near Stockholm had had an abortion. The eight-inch-long female foetus was wrapped in a sterile green cloth and delivered to a yellow brick outbuilding on the grounds of the National Biological Laboratory in north-west Stockholm. The lungs were removed, packed in ice and flown to the Wistar Institute.

The
The Wistar Institute in Philadelphia. Photograph: The Wistar Institute

Hayflick had been waiting months for this opportunity. These lungs would be the source of the new cells he needed to make antiviral vaccines. Viruses cant multiply outside living cells, and huge quantities of virus were needed to produce vaccines.

Now, at last, the lungs were here in his bustling second-floor lab, two purplish things floating in clear pink fluid in a glass bottle. They had been sent to Hayflick by a top virologist at the prestigious Karolinska Institute in Stockholm.

Hayflick knew that he was uniquely positioned to produce a long-lasting supply of these cells. He had spent the previous three years perfecting the procedure that would do it.

Hayflick took the lungs into a tiny room just off his lab what passed for a sterile area in 1962. He picked up a pair of tweezers, dipped them in alcohol and passed them through the flame of a Bunsen burner. He waited for them to cool and then, gently, one at a time, lifted the organs and placed them on a petri dish. Each was no larger than his thumb above the knuckle. He began carefully slicing them into innumerable pieces, each smaller than a pinhead.

Hayflick nudged the minute pieces of tissue into a wide-mouthed glass flask. The translucent pink fluid was full of digestive enzymes from slaughtered pigs. These biological jackhammers broke up the mortar between the lung cells, separating millions upon millions of them. Later, he transferred those cells into several flat-sided glass bottles and poured a nutritious solution over them. Hayflick then loaded the bottles on to a tray, and carried them into an incubation room where the temperature was a cosy 36C. He laid the bottles on their sides on a wooden shelf and closed the door carefully behind him. There the cells began to divide. He already had a name for them: WI-38.


The WI-38 cells that Hayflick launched that day were used to make vaccines that have been given to more than 300 million people half of them preschool children in the US. A copycat group of cells, developed using the method that Hayflick pioneered, has been used to make an additional 6bn doses of various vaccines.

Together these vaccines have protected people the world over from the gamut of viral illnesses: rubella, rabies, chickenpox, measles, polio, hepatitis A, shingles and adenovirus a respiratory infection that flourishes in situations where people live in close quarters. (Every US military recruit more than nine million of them since 1971 is given an adenovirus vaccine made using WI-38 cells.) In the US, a vaccine made in WI-38 cells that is still given to young children has wiped out homegrown rubella. It was developed at the Wistar Institute by Hayflicks colleague Stanley Plotkin, during a rubella epidemic that swept the country in 1964 and 1965.

The WI-38 cells are still in use today partly because Hayflick made such a large initial stock of them: some 800 tiny, wine-bottleshaped ampoules were frozen in the summer of 1962. When frozen, cells stop dividing, but then gamely begin replicating when they are thawed.Each glass vial that Hayflick froze contained between 1.5m and 2m cells. The cells in those vials had, on average, the capacity to divide about 40 more times. Early on, Hayflick determined that the newly derived cells in just one of his small glass lab bottles, if allowed to replicate until they died, would produce 20m tonnes of cells.In those 800 vials, he had created a supply of cells that for practical purposes was almost infinite.

In addition to their use in vaccine making, the WI-38 cells became the first normal cells available in virtually unlimited quantities to scientists probing the mysteries of cell biology. Because they were easily infected with human viruses, they became important to disease detectives tracking viruses in the 1960s, before more sophisticated technology came along.Biologists still reach for WI-38 cells when they need a normal cell to compare against a cancerous one, or to test the toxicity of new drugs. They are a workhorse of research into ageing, because they so reliably age and die in laboratory conditions. Original ampoules of WI-38 cells, and of polio vaccine made using them, are now part of the collection of the National Museum of American History.

But in the 1960s and 70s, a bitter feud broke out between Hayflick and the US government over who owned the cells.

Hayflick
Hayflick in the lab in the 1960s. Photograph: Courtesy of the University of Pennsylvania archives

As the importance of the WI-38 cells grew,Hayflick was only too happy to promote them. Human Cells Given Role in Vaccines, the New York Times proclaimed after the scientist spoke at a vaccine conference in 1966. The article quoted Hayflick explaining that his cells were cheaper, cleaner and safer than the animal cells then used in vaccine manufacture.

As his profile rose, Hayflick ran out of patience with Koprowski. The disconnect between his contributions and his treatment by the Wistar Institutes director had become too much to bear. Nine years after Koprowski hired him, Hayflick remained stuck as an associate member of the institute, in sharp contrast to many colleagues who had been made full members despite, to his mind, making contributions no greater than his own.

Hayflick began looking around. He applied for a position as a full professor of medical microbiology at Stanford University in Palo Alto, California. His application for the job was backed by a recommendation from a senior virologist who regarded his work as reliable, trustworthy and original. He was offered the post.

As Hayflicks departure approached, there was probably only one thing that concerned Koprowski: the fate of the hundreds of ampoules of WI-38 cells that were still stored in liquid nitrogen in the Wistar Institutes basement, under Hayflicks watchful eye. Hayflicks proprietary feelings about the cells were well known he once described them as like my children.

Koprowski had designs on the cells from the beginning. Nancy Pleibel, a lab technician who worked for Hayflick, recalls that more than once Koprowski had turned up in the lab within a day or two of Hayflick leaving on a trip, smiling and asking her for an ampoule of WI-38 cells. Politely but firmly, she refused his requests, explaining that only her boss could hand out WI-38 ampoules. After a while, Koprowski stopped asking.

Minutes from meetings of the Wistar Institutes board of managers in the early and mid-60s make clear that Koprowski tried repeatedly to cash in on Hayflicks human diploid cells(defined as cells that carry the normal complement of 46 chromosomes). The institute sought payment not only from Norden, a Missouri company that was interested in using WI-38 to develop a rabies vaccine, but also from Pfizer for the use of Hayflicks cells to make a measles vaccine, and from Wyeth, another Philadelphia-based drug manufacturer that by 1965 had used the WI-38 cells to make an adenovirus vaccine to protect US army recruits during basic training.

Koprowskis attempts to turn a profit with the WI-38 cells were far from successful. By 1965, the board of managers had appointed a special committee of lawyers and scientists to deal with problems in selling the Hayflick cells to industry. The only backing that the institute landed, according to budget documents from 1965 to 1967, was $5,000 in each of those years from Norden.

Today it seems incredible that an institution like the Wistar, full of eminent scientists, was so at sea when it came to profiting from unique and desirable cells produced under its roof. But in that era living things, such as the WI-38 cells, could not be patented. It would take a landmark supreme court decision in 1980 to change that.

However, what could be patented was a method of using the cells to produce a novel vaccine. Koprowski had already applied, back in 1964, for such a patent for another, improved rabies vaccine that he was developing using the WI-38 cells. Soon the Wistar Institute would apply for a patent on a method of making a rubella vaccine with the WI-38s, devised by another of its scientists, Stanley Plotkin.

If and when the rabies and rubella vaccine patents were granted, Koprowski would need accessto at least some of the original ampoules of WI-38 frozen in the Wistar Institute basement.Vaccine companies would want original ampoules full of the youngest cells, which could be expanded into a nearly endless supply.

By the autumn of 1967, Hayflick vaguely suspected that Koprowski intended the WI-38 cells to serve something more thanthe good of mankind. Hayflick believed that his boss hoped to turn any vaccines made with the cells into sources of cash, boosting the Wistar Institutes income and freeing him from fundraising duties that he detested and considered beneath him.

Transmission
Transmission electron micrograph of Rubella virions. Photograph: Science Photo Library

Hayflicks instincts were right. As 1967 drew to a close, a financial vice was tightening on Koprowski. While the Wistar Institute had remained solvent, it had never been flush with funds, especially after Koprowski blew through $271,506 to fund major renovations that were completed in 1959. By the mid-60s, his struggle to find cash not tied to specific grants was becoming desperate. Badly needed repairs to the roof and the air conditioning system were deferred.

In the autumn of 1967, when officials at the NIHs National Cancer Institute (NCI) learned that Hayflick would be moving to Stanford, they decided to take the production, storage, study and distribution to researchers of human diploid cells out of his hands. The NCI had been paying the Wistar Institute hundreds of thousands of dollars for Hayflick to produce and distribute the cells since 1962, shortly after his paper announcing his human diploid cell strains to the world had sent demand soaring. The agencys contract with the Wistar Institute had specified that the government would take ownership of the cells when the contract was terminated. Now, NIH officials set 1 January 1968 as the end date. The timing seemed right, and not only because of Hayflicks impending move. The sense at the NCI was that the demand for the WI-38 cells had been sated. Those scientists who wanted them, it seemed, had them by now, more than five years after Hayflick had first produced them. They were being used widely and had already been cited in scores of papers.

On 18 January 1968, several men travelled to the Wistar Institute to sort out the physical disposition of the WI-38 cells now that the contract had ended. Koprowski summoned Hayflick to meet with them. Also present were senior scientists from the American Type Culture Collection (ATCC). This independent, nonprofit organisation was the countrys highest-profile cell bank, and was often where biologists turned when they needed a particular type of cell for an experiment. According to records, the assembled men agreed that all but 20 of the roughly 375 remaining original ampoules of WI-38 cells would be transferred to the ATCC, which would maintain them, deeply frozen, on behalf of the NIH. Hayflick would be permitted to take 10 ampoules with him to Stanford, and the Wistar Institute would also be allowed to keep 10.

The group also decided that any use of the 355 original ampoules being transferred to the ATCC they were precious because the WI-38 cell populations in them had divided only eight times, and so could be expanded into untold billions of cells for vaccine making should be totally arrested. By this, they meant that there was to be no more thawing of the ampoules, no more planting of these young cells into lab bottles, and no more splitting of those bottles over and over to generate multitudes of cells at higher doubling levels for scientists to use. Scientists could use the older cells that were already in circulation. The remaining 355 original ampoules needed to be kept safely frozen at the ATCC until such time as companies began winning US licences to make WI-38based vaccines.

Some time during his last months at the Wistar Institute, Hayflick was working in one of the tiny sterile rooms that adjoined his lab. Plotkin squeezed through the door and pulled up the only chair. The two chatted for a while, then Plotkin showed Hayflick a document. It was a letter, on Wistar-headed paper, from Koprowski, written to a senior official at Burroughs Wellcome, the British pharmaceutical company. Koprowski was offering to provide to the company ample supplies of WI-38 cells, along with the recipe for making a vaccine with the cells and the virus itself, all in exchange for royalties.

Hayflicks suspicions had been confirmed. He was profoundly upset. He had spent the previous decade deriving the cells and opened up a new, important field in the study of cellular ageing. He had derived enough WI-38 cells to serve vaccine makers into the distant future and worked as hard as was humanly possible to win acceptance of the cells for vaccine making. In the process of all of this, he had been ridiculed and been forced to struggle for respect and validation.

This letter signalled that not only was he not valued but that he was also being sidelined in major decision-making and likely profit-making connected to the WI-38 cells. As Hayflick said, to have the vultures descend on what I had struggled so hard to give value to and [for them to] try to take it for their benefit I think that an average person would understand why I was, to put it mildly, concerned.

On or around 1 March when, under the January agreement, the ampoules were to have been moved from the Wistar Institute to the ATCC a specially outfitted station wagon arrived from Maryland, carrying the NIH project officer, Charles Boone, and John Shannon, the ATCCs curator of cell lines. Hayflick turned them away, saying he wasnt ready to hand over the cells because he had not prepared an inventory of them.

Not long after this, Hayflick, unobserved, visited the Wistar Institutes basement. There he packed every single one of the remaining original WI-38 ampoules 375 frozen vials: the largest stock of young WI-38 cells on earth into one or more portable liquid-nitrogen refrigerators and departed the premises. He left nothing behind not even the 10 ampoules that Koprowskis institute had been promised in the January agreement.

Hilary
Hilary Koprowski at work in his lab. Photograph: Yale Joel/The Life Picture Collection/Getty Images

Hayflick stored the frozen cells temporarily with a friend, a vaccinologist at the nearby Wyeth Laboratories who, from time to time, topped up the liquid nitrogen that kept the cells frozen. Hayflick says that he took the ampoules with the intention of keeping them only until the ownership of the cells could be properly sorted out. He believed that there were several potential stakeholders who might reasonably claim ownership: himself and his early collaborator at the Wistar Institute, the chromosome expert Paul Moorhead; the estate of the WI-38 foetus, by which he meant the WI-38 foetuss parents; the Wistar Institute; and, just possibly, the NIH. But he was not going to be so naive as to leave the cells in the NIHs possession while these matters were decided. If he did that, he was sure that he would never see them again.

In mid-1968, Hayflick left for his new job in California. Moving a family of seven 2,900 miles was no small undertaking. The Hayflicks split the travel. Ruth flew out to the San Francisco Bay Area with their two youngest daughters. Hayflick drove the three older children cross-country in their dark green Buick sedan. They drove west through Pittsburgh, stopped to see drag races in Joplin, Missouri, and then headed on to Arizona, where they gazed at the worlds best-preserved meteor crater and marvelled at the Grand Canyon. All along the way, some extra cargo travelled with them. Carefully strapped on the backseat beside his children was a liquid-nitrogen refrigerator stuffed with ampoules of WI-38.


Hayflicks flight with the cells would make him the target of a career-derailing investigation by the National Institutes of Health. Hayflick counter-sued eventually, in 1981, settling with the government. He was allowed to keep six original ampoules of the cells, along with $90,000 that he had earned by charging researchers and companies for them after he left the Wistar Institute. A letter from supporters published in the journal Science, described the happy outcome of Dr Hayflicks courageous, sometimes lonely, emotionally damaging and professionally destructive ordeal.

But just as the tug-of-war over ownership of the WI-38 cells peaked, profound changes occurred in attitudes and laws governing who could make money from biological inventions. In the space of a few years, biologists went from being expected to work only for their salaries and the greater good to being encouraged by universities and the government to commercialise their innovations for the benefit of the institutions, the US economy and themselves.

Although the WI-38 cellswere launched long before these changes took place and 18 years before the supreme court decreed that a living entity, such as a WI-38 cell, could be patenteda lot of money has been made from them. The drug company Merck, in particular, has made billions of dollars by using the WI-38 cells to make the rubella vaccine given to more than seven million American children each year. The Wistar Institute too enjoyed a handsome royalty stream from vaccines made by its scientists using the cells including a much-improved rabies vaccine that replaced sometimes dangerous injections. Cell banks today charge several hundred dollars for a tiny vial of the cells.

During the long battle for ownership of the WI-38 cells, Koprowski sent a Wistar scientist across the country to collect them from Hayflicks Stanford lab. But Hayflick refused to part with them. A second emissary was more successful, returning with the 10 ampoules originally allocated to the institute. But later, while the NIH was still asserting its title to WI-38, Koprowski seems to have given up. Perhaps this was because Hayflick was now so far away. Maybe it was because, despite his propensity for it, Koprowski actually disliked direct conflict. Possibly, it was because several companies already appeared to have adequate supplies of the youngest WI-38 ampoules. On the other hand, though, it might have been because Koprowski had finally realised just how persistent, obdurate and dedicated Hayflick could be.

This is an adapted extract from The Vaccine Race by Meredith Wadman, published by Doubleday on 9 February in the UK and in the US by Viking.

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