Viruses and Virus Nucleic Acid Contaminate Many Vaccines

Risks of cancer and creation of new pathogens should not be underplayed by regulators. Prof. Joe Cummins

Garbage viruses and DNA in vaccines

Vaccines are currently produced using fertilized chicken eggs, cell culture or a combination of egg and cell culture. An ‘attenuated’ vaccine is created from a pathogen by reducing its virulence, but still keeping it viable, in contrast to those produced by ‘killing’ the virus (inactivated vaccine). Inactivation is done by selecting non-pathogenic strains of the pathogen after treatment such as heat or cold culture, or targeted deletion of virulence genes.

Many live attenuated vaccines are produced using cell culture. A number of such   vaccines have been found to contain not only the live attenuated viral pathogen but also contaminating viruses or viral nucleic acid [1]. These contaminants are garbage, and people administering such vaccines should inform patients of potential risks associated with the garbage. Recently, the United States Food and Drug Administration (FDA) acknowledged the contamination of the live attenuated rotavirus vaccine (to prevent traveller’s diarrhoea) and suspended the vaccine, but later decided that the benefits of the vaccination outweigh potential contamination risks [2].  The FDA opinion is premature because the circovirus contaminating the vaccine is active in replication, transcription and translation of viral genes and able to produce toxic products. Contaminated vaccines are not isolated cases, they are widespread.

Lessons from SV40 contaminated vaccines

Simian virus 40 (SV40) is a monkey virus inadvertently administered to human populations in contaminated vaccines produced in SV40-infected monkey cells. Molecular biology and epidemiological studies suggest that SV40 may be contagiously transmitted in humans by horizontal infection, independently of the earlier administration of SV40-contaminated vaccines. In humans, SV40 has been found associated at high prevalence with specific tumour types such as brain and bone tumours, mesotheliomas and lymphomas and with kidney diseases [3]. SV40 was discovered as a contaminant of poliovirus vaccine lots distributed to millions of individuals in the United States between 1955 and 1963; and contaminated vaccine batches were later circulated worldwide. After SV40 was observed to cause animal and human cell transformations in culture, and tumour formations in animals, researchers began to search for SV40 in human cancers [4]. For example, a 2005 study undertaken in Costa-Rica showed that SV40 is significantly associated with cancers of the immune system [5]. US FDA acknowledges that the SV 40 virus (simian virus 40 from monkey kidney cells) was in the early polio vaccines and its risks [6]: “The experience in the early 1960s with SV40 contamination of poliovirus and adenovirus vaccines and the continuing questions regarding whether SV40 could be responsible for some human neoplasms [cancers] underscores the importance of keeping viral vaccines free of adventitious agents. “ (See also Flu Vaccines and the Risk of Cancer, SiS 44 [7]). SV40 contamination of  polio vaccines is an old lesson that seems to have been ignored  in the current rush to profit from manufacturing  vaccines.

Numerous vaccines for humans are contaminated

There are numerous cases of documented contaminated vaccines intended for humans [1]. Measles vaccine Attenuvax grown in chicken embryo fibroblast cells was contaminated with Avian leucosis (myeloid leucosis cancer virus) and avian endogenous retrovirus. Yellow fever vaccine YFvax  grown in chicken embryo fibroblast cells was contaminated  with  avian endogenous retrovirus. Herpes 3 vaccine Varivax grown in MRC-5 human cells from aborted foetuses was contaminated with human endogenous retrovirus K.  Rota virus vaccine Rotarix  grown in Vero E6 (African green monkey ) cells was contaminated with with porcine circovirus 1 and porcine circovirus 2. Rotavirus Rotateq vaccine grown in Vero (African monkey) cells had Baboon endogenous retrovirus as contaminant.  Measles mumps vaccine MMR II grown in chicken fibroblast cells had Avian endogenous retrovirus and human endogenous retrovirus K as contaminants;  and Rubella vaccine grown in  WI-38 human diploid lung fibroblast cells was contaminated with Human endogenous retrovirus K.  Rubella vaccine meruvax II grown in WI-38 human lung fibroblast cells contained human endogenous retrovirus-K.

Veterinary vaccines are similarly contaminated. The genomes of all animal species are colonized by endogenous retroviruses (ERVs). Although most ERVs have accumulated defects that render them incapable of replication, fully infectious ERVs have been identified in various mammals.  A feline infectious ERV (RD-114w) was isolated from many live attenuated  vaccines for pets. Isolation of RD-114 was done independently in two laboratories using different detection strategies and from vaccines for both cats and dogs commercially available in Japan or the UK. The study shows that the methods currently employed to screen veterinary vaccines for retroviruses are inadequate and should be re-evaluated [8]. Tests of veterinary   vaccines for viral contamination in Hungary found that a torquetenovirus (TTV), a very small circular single stranded DNA virus, was present in many vaccines including avian vaccines. The presence of any extraneous agent may have a significant impact on the safety of the vaccine [9].

A rogues’ gallery of vaccine contaminating viruses and DNA

Avian leukosis (myeloid leukosis cancer virus)

Avian leukosis virus (ALV-J) appears to be a recombinant of an exogenous avian leukosis virus (ALV) with an envelope (env) gene probably originating from an endogenous (subgroup E) ALV. ALV-J can infect cell cultures from other avian species, but not mammalian cells. No genetically resistant meat-type strain of chickens has been found to date. Commercial Leghorn chickens appear to be resistant to tumour development, but they may be susceptible to infection. Most tumours associated with ALV-J infection are expressed as myeloblastomas or myelocytomas [10]. Even though the bird cancer virus does not appear to infect mammals, the persistent exposure of young human may select mutations of the virus that are virulent in people; and virulent recombinants can always be created with endogenous human viruses.

Avian endogenous retrovirus

Avian endogenous retrovirus (AER) are a highly diverse group comprising many inserts into the chicken genome. There are three families of such endogenous retroviruses, related respectively to avian sarcoma or leukosis cancer virus, mouse leukemia viruses, and human endogenous retroviruses. Most of the AER are dormant in the chicken chromosomes, but several are active and capable of making RNA transcripts [11]. The active transcripts may replicate by reverse transcription and recombine with related viruses.

Human endogenous retrovirus K

Human endogenous retroviruses (HERVs) are suspects in some autoimmune diseases, in particular, multiple sclerosis; a member of the family of human endogenous retrovirus W has been identified as “MS-associated retrovirus” (MSRV). HERVs comprise nearly 8 percent of the human genome, with 98 000 elements and fragments [11]; all appear to be defective, containing nonsense mutations or major deletions, and cannot produce infectious virus particles. Most are remnants of viruses that integrated many millions of years ago. However, one family – HERV-K (comprising less than 1 percent of HERV elements) - have been active since the divergence of humans and chimpanzees, and is one of the most studied. There are indications it has even been active in the past few hundred thousand years, as some human individuals carry more copies of the virus. The lack of elements with a full coding potential within the published human genome sequence suggests that the family is less likely to be active at present [6]. HERV-K contaminants in vaccines should not be considered innocuous as they may recombine with related viruses or with viral sequences in the human chromosome.

Baboon endogenous retrovirus

Baboon endogenous retrovirus (BERV) is a inactivated retroviral sequence. BERVs are also found in the African green monkey [12].  BERV circulating in the bloodstream of humans could conceivably mutate and recombine to form a virus that could spread rapidly in the human population because the virus is new to the immune repertoire of the human.

Feline infectious ERV (RD-114)

An infectious endogenous retrovirus was discovered in live attenuated vaccine for cats and dogs. EVR RD-114 is related to other oncogenic virus such as feline leuekemia virus and mouse leukemia virus 83 [12].

Porcine circovirus 1 and porcine circovirus 2

The pig circoviruses are small circular single stranded DNA viruses. Type 1 virus does not cause illness in pigs while type 3 virus causes a serious wasting disease of young pigs. The viruses are frequently found infecting mammalian cell lines.  Circovirus type 1 and type 2 infect many human cell types. Type 1 virus proliferates without causing distinct cell damage while type 2 virus does [13]. Type 2 virus causes cytoskeleton rearrangements in dendritic cells, leading to immunosuppression [14]. Porcine circovirus is lodged in the cell nucleus where it is replicated. Replication is by a rolling circle mechanism where the single stranded viral chromosomes are rolled off a double stranded replicative master. The virus is so small that it only has room for a few genes including two genes for initiating DNA replication along with genes for nuclear localization and viral coat protein and a few genes for virulence [15].  The host cell nucleus provides the enzymes for DNA replication [16].

Torquetenovirus (TTV)

Torquetenoviruses (TTVs) are vertebrate infecting, single-stranded circular DNA viruses. Two genetically distinct TTV groups (TTV1 and TTV2) infect swine worldwide with high prevalence. Currently, swine TTVs are considered non-pathogenic, although TTV2 has been linked to post-weaning multisystemic wasting syndrome, a porcine circovirus disease TTV replicates similarly to the circovirus but is much smaller than the circovirus [17]. TTV is often presumed to be non-pathogenic, and is distributed widely among mammals including humans.  TTV infection is widely dispersed in the human population and the virus has been found to accumulate in the central nervous system and implicated in dementia [18].  Children with recurrent pneumonia have been found to lack ciliary motility associated with high level infection of ciliarycells with TTV [19].

To conclude

Human and veterinary vaccines have been found contaminated with wide array of viruses that are deemed harmless or less risky than the attenuated live virus of the vaccine.  These contaminating garbage viruses are nowhere near as well investigated than they should have been prior to the commercialization of the vaccines. The contaminating garbage viruses are deemed harmless because they do not elicit sera conversion (production of antibody) even though the garbage viruses frequently produce proteins that are toxic in specific tissues.  The contaminating garbage vaccines are actively cytotoxic in some cases, and potentially so in other cases by mutation or recombination to create new retroviruses that are life threatening. Among the garbage viruses, the small circular single stranded DNA viruses deserve special attention as they are so widespread in the human and animal populations. Such widespread dispersal of TTV and circoviruses could cause disaster.  The first step in dealing with the garbage viruses is to provide informed consent to those being vaccinated with contaminated vaccines. The second is to carry out post-release monitoring for potential hazards from mutation and recombination, as highlighted in this article.

Article first published 13/12/10

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References

1. Victoria JG, Wang C, Jones MS, Jaing C, McLoughlin K, Gardner S, Delwart EL.Viral nucleic acids in live-attenuated vaccines: detection of minority variants and an adventitious virus. J Virol 2010, 84(12), 6033-40
2. Kuehn BM. FDA: Benefits of rotavirus vaccination outweigh potential contamination risk. JAMA 2010, 304(1), 30-1.
3. Martini F, Corallini A, Balatti V, Sabbioni S, Pancaldi C, Tognon M. Simian virus 40 in humans. Infect Agent Cancer 2007, 2, 13.
4. Lowe DB, Shearer MH, Jumper CA, Kennedy RC. SV40 association with human malignancies and mechanisms of tumor immunity by large tumor antigen. Cell Mol Life Sci 2007, 64(7-8), 803-14.
5. Meneses A, Lopez-Terrada D, Zanwar P, Killen DE, Monterroso V, Butel JS, Vilchez RA. Lymphoproliferative disorders in Costa Rica and simian virus 40. Haematologica 2005, 90(12), 1635-42.
6. Wikipedia Human endogenous retroviruses (HERVs) are suspected of involvement in some autoimmune diseases, in particular with multiple sclerosis. 2010 http://en.wikipedia.org/wiki/Endogenous_retrovirus
7. Tenpenny S. Flu vaccines and risk of cancer  Science in Society 44, 25-26, 2009.
8. Miyazawa T, Yoshikawa R, Golder M, Okada M, Stewart H, Palmarini M.Isolation of an infectious endogenous retrovirus in a proportion of live attenuated vaccines for pets. J Virol 2010, 84(7), 3690-4.
9. Kulcsar G, Farsang A, Soos T.Testing for viral contaminants of veterinary vaccines in Hungary. Biologicals 2010, 38(3), 346-9.
10. Zavala G. An overview of myeloid leukosisin meat-type chickens Technical News  Special Technical Bulletin Jan. 1998 http://birdflubook.com/resources/zavala1.pdf
11. Boristenko L. Avian endogenous retroviruses  Folio Biologica (Praha) 2003, 49, 177-82
12. Mang R, Goudsmit J, van der Kuyl AC Novel endogenous type C retrovirus in baboons: complete sequence, providing evidence for baboon endogenous virus gag-pol ancestry. J Virol 1999, 73(8), 7021-6.
13. Hattermann K, Roedner C, Schmitt C, Finsterbusch T, Steinfeldt T, Mankertz A. Infection studies on human cell lines with porcine circovirus type 1 and porcine circovirus type 2 Xenotransplantation 2004, 11(3), 284-94.
14. Balmelli C, Steiner E, Moulin H, Peduto N, Herrmann B, Summerfield A, McCullough K Porcine circovirus type 2 DNA influences cytoskeleton rearrangements in plasmacytoid and monocyte-derived dendritic cells. Immunology 2010 Sep 14. doi: 10.1111/j.1365-2567.2010.03339.x. [Epub ahead of print]
15. Faurez F, Dory D, Grasland B, Jestin A. Replication of porcine circoviruses. Virol J 2009, 6, 60. Review.
16. Gassmann M, Focher F, Buhk HJ, Ferrari E, Spadari S, Hübscher U. Replication of single-stranded porcine circovirus DNA by DNA polymerases alpha and delta. Biochim Biophys Acta 1988, 951(2-3), 280-9.
17. Kekarainen T, Martínez-Guinó L, Segalés J. Swine torque teno virus detection in pig commercial vaccines, enzymes for laboratory use and human drugs containing components of porcine origin. J Gen Virol 2009, 90(3), 648-53.
18. Maggi F, Bendinelli M. Human anelloviruses and the central nervous system. Rev Med Virol 2010, 20(6), 392-407.
19. Pifferi M, Maggi F, Di Cristofano C, Cangiotti AM, Nelli LC, Bevilacqua G, Macchia P, Bendinelli M, Boner AL. Torquetenovirus infection and ciliary dysmotility in children with recurrent pneumonia. Pediatr Infect Dis J 2008, 27(5), 413-8.