title: Direct evidence of extensive diversity of HIV-1 in Kinshasa by 1960.
abstract: Human immunodeficiency virus type 1 (HIV-1) sequences that pre-date the recognition of AIDS are critical to defining the time of origin and the timescale of virus evolution. A viral sequence from 1959 (ZR59) is the oldest known HIV-1 infection. Other historically documented sequences, important calibration points to convert evolutionary distance into time, are lacking, however; ZR59 is the only one sampled before 1976. Here we report the amplification and characterization of viral sequences from a Bouin's-fixed paraffin-embedded lymph node biopsy specimen obtained in 1960 from an adult female in Léopoldville, Belgian Congo (now Kinshasa, Democratic Republic of the Congo (DRC)), and we use them to conduct the first comparative evolutionary genetic study of early pre-AIDS epidemic HIV-1 group M viruses. Phylogenetic analyses position this viral sequence (DRC60) closest to the ancestral node of subtype A (excluding A2). Relaxed molecular clock analyses incorporating DRC60 and ZR59 date the most recent common ancestor of the M group to near the beginning of the twentieth century. The sizeable genetic distance between DRC60 and ZR59 directly demonstrates that diversification of HIV-1 in west-central Africa occurred long before the recognized AIDS pandemic. The recovery of viral gene sequences from decades-old paraffin-embedded tissues opens the door to a detailed palaeovirological investigation of the evolutionary history of HIV-1 that is not accessible by other methods.
authors.affiliation: Ecology and Evolutionary Biology, University of Arizona, Tucson, Arizona 85721, USA. worobey@email.arizona.edu
authors.forename: Michael
authors.initials: M
authors.lastname: Worobey
authors.forename: Marlea
authors.initials: M
authors.lastname: Gemmel
authors.forename: Dirk E
authors.initials: DE
authors.lastname: Teuwen
authors.forename: Tamara
authors.initials: T
authors.lastname: Haselkorn
authors.forename: Kevin
authors.initials: K
authors.lastname: Kunstman
authors.forename: Michael
authors.initials: M
authors.lastname: Bunce
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Muyembe
authors.forename: Jean-Marie M
authors.initials: JM
authors.lastname: Kabongo
authors.forename: Raphaël M
authors.initials: RM
authors.lastname: Kalengayi
authors.forename: Eric
authors.initials: E
authors.lastname: Van Marck
authors.forename: M Thomas P
authors.initials: MT
authors.lastname: Gilbert
authors.forename: Steven M
authors.initials: SM
authors.lastname: Wolinsky
mesh_terms: D000328:Adult; D002170:Canada; D015023:Democratic Republic of the Congo / Q000453:epidemiology; D019143:Evolution, Molecular; D005260:Female; D014644:Genetic Variation / Q000235:genetics*; D015658:HIV Infections / Q000453:epidemiology* / Q000473:pathology / Q000821:virology*; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D049673:History, 20th Century; D006801:Humans; D008297:Male; D008867:Microtomy; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA