title: Mitochondrial DNA and retroviral RNA analyses of archival oral polio vaccine (OPV CHAT) materials: evidence of macaque nuclear sequences confirms substrate identity.
abstract: Inoculation of live experimental oral poliovirus vaccines (OPV CHAT) during the 1950s in central Africa has been proposed to account for the introduction of HIV into human populations. For this to have occurred, it would have been necessary for chimpanzee rather than macaque kidney epithelial cells to have been included in the preparation of early OPV materials. Theoretically, this could have led to contamination with a progenitor of HIV-1 derived from a related simian immunodeficiency virus of chimpanzees (SIVCPZ). In this article we present further detailed analyses of two samples of OPV, CHAT 10A-11 and CHAT 6039/Yugo, which were used in early human trials of poliovirus vaccination. Recovery of poliovirus by culture techniques confirmed the biological viability of the vaccines and sequence analysis of poliovirus RNA specifically identified the presence of the CHAT strain. Independent nested sets of oligonucleotide primers specific for HIV-1/SIVCPZ and HIV-2/SIVMAC/SIVSM phylogenetic lineages, respectively, indicated no evidence of HIV/SIV RNA in either vaccine preparation, at a sensitivity of 100 RNA equivalents/ml. Analysis of cellular substrate by the amplification of two distinct regions of mitochondrial DNA (D-loop control region and 12S ribosomal sequences) revealed no evidence of chimpanzee cellular sequences. However, this approach positively identified rhesus and cynomolgus macaque DNA for the CHAT 10A-11 and CHAT 6039/Yugo vaccine preparations, respectively. Analysis of multiple clones of mtDNA 12S rDNA indicated a relatively high number of nuclear mitochondrial DNA sequences (numts) in the CHAT 10A-11 material, but confirmed the macaque origin of cellular substrate used in vaccine preparation. These data reinforce earlier findings on this topic providing no evidence to support the contention that poliovirus vaccination was responsible for the introduction of HIV into humans and sparking the AIDS pandemic.
authors.affiliation: Division of Retrovirology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. nberry@nibsc.ac.uk
authors.forename: Neil
authors.initials: N
authors.lastname: Berry
authors.forename: Adrian
authors.initials: A
authors.lastname: Jenkins
authors.forename: Javier
authors.initials: J
authors.lastname: Martin
authors.forename: Clare
authors.initials: C
authors.lastname: Davis
authors.forename: David
authors.initials: D
authors.lastname: Wood
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Schild
authors.forename: Margareta
authors.initials: M
authors.lastname: Bottiger
authors.forename: Harvey
authors.initials: H
authors.lastname: Holmes
authors.forename: Philip
authors.initials: P
authors.lastname: Minor
authors.forename: Neil
authors.initials: N
authors.lastname: Almond
mesh_terms: D000818:Animals; D001483:Base Sequence; D018070:Biological Specimen Banks; D002460:Cell Line; D002522:Chlorocebus aethiops; D004272:DNA, Mitochondrial / Q000032:analysis*; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D006801:Humans; D008251:Macaca; D051379:Mice; D008969:Molecular Sequence Data; D002679:Pan troglodytes; D010215:Papio; D010802:Phylogeny; D011055:Poliovirus Vaccine, Oral / Q000235:genetics*; D012335:RNA, Ribosomal / Q000032:analysis* / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods
chemical_list: D004272:DNA, Mitochondrial; D004275:DNA, Ribosomal; D011055:Poliovirus Vaccine, Oral; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S