Simian Virus 40, Poliovirus Vaccines, and Human Cancer: Research Progress Versus Media and Public Interests J.S

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Simian virus 40, poliovirus vaccines, and human cancer: research progress versus media and public interests J.S. Butel1

From 1955 through early 1963, millions of people were inadvertently exposed to simian virus 40 (SV40) as a contaminant of poliovirus vaccines; the virus had been present in the monkey kidney cultures used to prepare the vaccines and had escaped detection. SV40 was discovered in 1960 and subsequently eliminated from poliovirus vaccines. This article reviews current knowledge about SV40 and considers public responses to reports in the media. SV40 is a potent tumour virus with broad tissue tropism that induces tumours in rodents and transforms cultured cells from many species. It is also an important laboratory model for basic studies of molecular processes in eukaryotic cells and mechanisms of neoplastic transformation. SV40 neutralizing antibodies have been detected in individuals not exposed to contaminated poliovirus vaccines. There have been many reports of detection of SV40 DNA in human tumours, especially mesotheliomas, brain tumours and osteosarcomas; and DNA sequence analyses have ruled out the possibility that the viral DNA in tumours was due to laboratory contamination or that the virus had been misidentified. However, additional studies are necessary to prove that SV40 is the cause of certain human cancers. A recently published review article evaluated the status of the field and received much media attention. The public response emphasized that there is great interest in the possibility of health risks today from vaccinations received in the past.

Keywords: drug contamination; mass media; poliovirus vaccine, adverse effects; Polyomavirus maccacae; public opinion.

Voir page 197 le reÂsumeÂ en francËais. En la paÂ gina 197 figura un resumen en espanÄ ol.

Simian virus 40 (SV40) is a polyomavirus of rhesus T-antigen (T-ag), a nonstructural protein essential for macaque origin (1±3). It was present but unrecog- viral replication (1, 2, 11). Fundamental to its driving nizedin the monkey kidney cell cultures usedto effects on the cell cycle, T-ag complexes with and prepare both the inactivatedandlive attenuated functionally inactivates cellular tumour suppressor poliovirus vaccines, as well as several other viral proteins (p53 andpRb), as well as other members of vaccines (4). Consequently, between 1955 andthe the pRb family (p107 andp130) ( 1, 12, 13). SV40 has early part of 1963, millions of people were also provided a facile and rewarding model for inadvertently exposed to live SV40 when they were molecular biology studies of eukaryotic cell processes administeredSV40-contaminatedvaccines ( 5±8). (1, 2); its genome was the first eukaryotic viral The major sources of exposure were poliovirus genome to be sequencedin its entirety. vaccines; not only was SV40 present in early lots of Although it was assumedfor many years that live poliovirus vaccine, but some infectious SV40 there was a single strain of SV40 andthat all isolates survivedthe inactivation treatments usedto prepare were identical, recent observations have refuted this the killedvaccine. As far as is known, no con- (2). Although available evidence indicates that there is taminatedpoliovirus vaccine has been usedsince a single serotype, sequence analyses have revealed early 1963. that multiple genetic strains exist. The viral regulatory Following its discovery in 1960, SV40 became region differs among isolates, with most laboratory- the object of intense investigation. It was foundto adapted strains containing a duplication in the possess potent oncogenic potential, having the ability enhancer that is usually lacking in fresh isolates from to induce tumours in rodents, to transform cells in monkeys andin DNA associatedwith human tissue culture (1±3, 9), andto promote tumour tumours (14±20). There is also a variable domain at formation in transgenic mice (10). The major the extreme C-terminus of T-ag that differs among transforming protein of SV40 was identified as large viral isolates (14, 18±20). These sequence differences provide a means of identifying viral strains and helped allay concerns that human tumour-associatedse- quences might have been due to laboratory contam- 1 Distinguished Service Professor and Chair, Department of Molecular ination. Virology and Microbiology, Baylor College of Medicine, Houston, SV40 infections in monkeys are generally TX 77030-3498, USA. asymptomatic. However, if the host's immune Ref. No. 0245

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system is compromised, SV40 disease may develop C-terminus of the T-ag gene andby the presence of (2). In macaques infectedwith simian immunodefi- a simple, nonduplicated enhancer in the regulatory ciency virus, SV40 lesions have been observedin region. Limitations of sample availability have many tissues, including brain, lung, kidney, lymph generally precluded studies of the integration state node and spleen (14±16, 21). Some animals devel- of viral DNA in the tumours andquantification of opedSV40 meningoencephalitis andothers the genome copies of the viral DNA present. demyelinating disease, progressive multifocal leuko- A recently publishedreview article evaluated encephalopathy. These observations indicate that the status of knowledge about the cell and molecular SV40 can be neurotropic andproduceinfections that biology of SV40 andthe implications for human involve the central nervous system. infections anddisease( 2). Although the data suggest Following the discovery of SV40, the conven- that SV40 DNA is present in some human tumours tional wisdom was that it was unable to establish andthat the virus probably has a causative role in human infections andwas, therefore, harmless. some cancers, more studies are necessary to build However, in studies dating back to the 1970s, SV40 definitive proof of etiology. The association of SV40 neutralizing antibodies have been detected in sera with human disease, if proven, would provide the from individuals born after the use of contaminated opportunity for new approaches to the diagnosis, vaccines, as well as in sera collectedbefore vaccines treatment, andprevention of human cancer. That, I were available (2). The low prevalence of serum believe, is the significance of this research, rather than antibodies (2±13%) suggested that there were the historical linkage of SV40 to the poliovirus sources of exposure to SV40, or a cross-reacting vaccines. However, media reports focused on the virus, other than the vaccines. In a recent study of possible connection between the contaminated hospitalizedchildrenborn from 1980 through 1995, vaccines andhuman cancer development. There SV40 neutralizing antibodies were detected in about was clear public interest in whether people who have 6% of unselectedsera ( 22). Tests were done on receivedthe contaminatedpoliovirus vaccines are at archival kidney tissue samples from 13 antibody- higher risk of getting cancer andwhether current positive patients andSV40 DNA was amplifiedby poliovirus vaccines are safe to give to children. The polymerase chain reaction (PCR) from specimens explanations that SV40 was once a contaminant of taken from four individuals, confirming that SV40 is poliovirus vaccines andwas now being detectedin present in children today (23). The source(s) of human tumours, but that we cannot say whether the contemporary SV40 infections is (are) unknown, but virus came from the oldvaccines or that the virus it is presumedthat the virus is being transmitted positively causedthe cancers, are subtle distinctions among humans. These serological andmolecular that failedto be includedin most reports. findings, coupled with numerous tissue culture This experience clearly shows that the public studies that have demonstrated that SV40 can has concerns about possible delayed side-effects replicate in human cells (2), prove that SV40 is able from vaccines andhas great interest in relatedhealth to infect humans. These data suggest, additionally, reports (Table 1). It has been estimatedthat there are that SV40 shouldbe consideredinfectious for both now about 15 000 health sites on the Internet, monkeys andhumans. providing new means for rapid dissemination of both SV40 DNA has been detected in human accurate andinaccurate information. It is fair to tumours in numerous independent studies (2, 24), assume that events similar to the poliovirus vaccine/ with the most commonly involvedcancers being cancer scare couldoccur with respect to other paediatric and adult brain tumours, mesotheliomas vaccines at some future time. It wouldtherefore be andosteosarcomas. Expression of T-ag, the SV40 advisable for researchers and public health officials to oncoprotein, has been observedin some tumours. be preparedfor such an occurrence. n Recent studies using PCR and DNA sequence technologies have substantiatedearlier reports dating from the mid-1970s of the presence of SV40 in Table 1. Reports about vaccine side-effects: human cancer tissue. Importantly, many of the SV40- public concerns positive tumours in recent studies have been from

patients too young to have been exposedto SV40- . The public has great interest in possible delayed health risks contaminatedpoliovirus vaccines. Evidence that due to vaccination authentic SV40 was detected includes the following . Modern communications allow rapid and widespread types of observations: four separatedregions of the dissemination of media reports viral genome have been PCR-amplifiedfrom . Some public suspicion exists about vaccine safety tumours; sequence analyses of PCR products have . Experts should present accurate information and interpreta- confirmedmatches with the SV40 DNA sequence; tions in response to media reports andviable SV40 has been isolatedfrom one . Science may not be the main message in media reports; paediatric brain tumour. Tumour-associated viral scientific subtleties get lost sequences have been distinguished from laboratory . Reporters have a responsibility to convey health-related strains of SV40 by variation in the sequence at the stories accurately

196 Bulletin of the World Health Organization, 2000, 78 (2) Simian virus 40, poliovirus vaccines, and human cancer

ReÂ sumeÂ SV40, vaccins antipoliomyeÂ litiques et cancer : les progreÁ s de la recherche et l'inteÂreÃt que leur portent les meÂ dias et le grand public Entre 1955 et 1963, des millions de personnes ont eÂteÂ dirigeÂs contre le SV40 chez des sujets qui n'avaient pas exposeÂ es par meÂ garde au SV40 (« simian virus 40 » des eÂteÂ exposeÂs aÁ des vaccins antipoliomyeÂ litiques conta- Anglo-Saxons) qui contaminait les vaccins antipoliomyeÂ- mineÂs. L'ADN du SV40 a eÂteÂaÁ de nombreuses reprises litiques ± ce virus ayant eÂteÂ preÂsent dans les cultures de deÂtecteÂ dans des tumeurs chez l'homme, surtout dans reins de singes utiliseÂ es pour preÂparer le vaccin et ayant des meÂsotheÂliomes, des tumeurs ceÂreÂbrales et des eÂchappeÂaÁladeÂtection. Le SV40 a eÂteÂdeÂ couvert en 1960 osteÂosarcomes ; et l'analyse des seÂquences de cet ADN a et a eÂteÂeÂlimineÂ par la suite des vaccins antipoliomyeÂ- exclu la possibiliteÂ d'une contamination de laboratoire litiques. Dans cet article, on fait le point des pour expliquer la preÂsence de cet ADN viral dans les connaissances actuelles concernant le SV40 et on tumeurs, ou d'une erreur d'identification du virus. examine les reÂ ponses du grand public aÁ l'eÂ cho qu'elles Toutefois, des eÂtudes suppleÂmentaires sont neÂcessaires ont eu dans les meÂdias. Le SV40 est un virus oncogeÁne pour prouver que le SV40 est bien aÁ l'origine de certains puissant ayant un important tropisme tissulaire qui induit cancers chez l'homme. Un article reÂ cemment publieÂ a fait des tumeurs chez les rongeurs et transforme les cultures le point de la question et a beaucoup retenu l'attention cellulaires de nombreuses espeÁces. C'est eÂgalement un des meÂdias. La reÂponse du grand public a souligneÂle modeÁle de laboratoire preÂcieux pour l'eÂtude fondamen- grand inteÂreÃt porteÂ au fait que l'on puisse courir des tale des processus moleÂ culaires en jeu dans les cellules risques aujourd'hui aÁ cause de vaccinations recËues dans eucaryotes et les meÂcanismes de transformation le passeÂ. neÂoplasique. On a deÂtecteÂ des anticorps neutralisants

Resumen Virus sõÂmico 40, vacunas contra el poliovirus y caÂ ncer humano: avances de las investigaciones e intereÂ s de los media y el puÂ blico Desde 1955 hasta principios de 1963, millones de anticuerpos neutralizantes del SV40 en personas no personas se vieron expuestas accidentalmente al virus expuestas a vacunas contra el poliovirus contaminadas. sõÂmico 40 (SV40), contaminante de las vacunas contra el Se han referido casos de deteccioÂ n de ADN SV40 en poliovirus. El virus habõÂa pasado desapercibido en los tumores humanos, en particular en mesoteliomas, cultivos de ceÂ lulas renales de mono utilizados para tumores cerebrales y osteosarcomas, y los anaÂ lisis de la elaborar las vacunas. El SV40 fue descubierto en 1960, secuencia del ADN tumoral han descartado la posibilidad tras lo cual fue eliminado de las vacunas contra el de que el ADN võÂrico de los tumores se deba a poliovirus. En este artõÂculo se examinan los conocimien- contaminacioÂ n de laboratorio o a errores de identifica- tos actuales sobre el SV40 y se considera la respuesta cioÂ n del virus. Sin embargo, es necesario realizar estudios puÂ blica a las noticias aparecidas en los medios de adicionales para demostrar que el SV40 provoca difusioÂ n. El SV40 es un potente virus tumoral con amplio determinados caÂ nceres humanos. Se ha analizado el tropismo tisular, que induce la aparicioÂ n de tumores en estado de la cuestioÂ n en un estudio crõÂtico publicado roedores y transforma las ceÂlulas de cultivo de muchas recientemente que ha atraõÂdo la atencioÂ n de los medios especies. TambieÂn es un importante modelo de de comunicacioÂ n. La respuesta puÂ blica ha puesto de laboratorio para realizar estudios baÂ sicos sobre procesos relieve el gran intereÂs que despierta la posibilidad de que moleculares en ceÂlulas eucariotas y sobre los mecanis- algunos riesgos sanitarios de hoy esteÂn asociados a mos de transformacioÂ n neoplaÂ sica. Se han detectado vacunas administradas en el pasado.

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