title: HIV-1 Integrates Widely throughout the Genome of the Human Blood Fluke Schistosoma mansoni.
abstract: Schistosomiasis is the most important helminthic disease of humanity in terms of morbidity and mortality. Facile manipulation of schistosomes using lentiviruses would enable advances in functional genomics in these and related neglected tropical diseases pathogens including tapeworms, and including their non-dividing cells. Such approaches have hitherto been unavailable. Blood stream forms of the human blood fluke, Schistosoma mansoni, the causative agent of the hepatointestinal schistosomiasis, were infected with the human HIV-1 isolate NL4-3 pseudotyped with vesicular stomatitis virus glycoprotein. The appearance of strong stop and positive strand cDNAs indicated that virions fused to schistosome cells, the nucleocapsid internalized and the RNA genome reverse transcribed. Anchored PCR analysis, sequencing HIV-1-specific anchored Illumina libraries and Whole Genome Sequencing (WGS) of schistosomes confirmed chromosomal integration; >8,000 integrations were mapped, distributed throughout the eight pairs of chromosomes including the sex chromosomes. The rate of integrations in the genome exceeded five per 1,000 kb and HIV-1 integrated into protein-encoding loci and elsewhere with integration bias dissimilar to that of human T cells. We estimated ~ 2,100 integrations per schistosomulum based on WGS, i.e. about two or three events per cell, comparable to integration rates in human cells. Accomplishment in schistosomes of post-entry processes essential for HIV-1replication, including integrase-catalyzed integration, was remarkable given the phylogenetic distance between schistosomes and primates, the natural hosts of the genus Lentivirus. These enigmatic findings revealed that HIV-1 was active within cells of S. mansoni, and provided the first demonstration that HIV-1 can integrate into the genome of an invertebrate.
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.|Department of Microbiology, Faculty of Science, Mahidol University, Phyathai, Rachthewee, Bangkok.|Department of Pathology, Faculty of Medicine, Khon Kaen University, Muang Khon Kaen, Thailand.
authors.forename: Sutas
authors.initials: S
authors.lastname: Suttiprapa
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.|Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Rinaldi
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.|Biodiversity Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Isheng J
authors.initials: IJ
authors.lastname: Tsai
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.
authors.forename: Victoria H
authors.initials: VH
authors.lastname: Mann
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.
authors.forename: Larisa
authors.initials: L
authors.lastname: Dubrovsky
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.|Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, Gansu, The People's Republic of China.
authors.forename: Hong-Bin
authors.initials: HB
authors.lastname: Yan
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Nancy
authors.identifier: http://orcid.org/0000-0003-2608-7151
authors.initials: N
authors.lastname: Holroyd
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huckvale
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Caroline
authors.identifier: http://orcid.org/0000-0002-0148-0603
authors.initials: C
authors.lastname: Durrant
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Anna V
authors.initials: AV
authors.lastname: Protasio
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Pushkarsky
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.|National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Sergey
authors.initials: S
authors.lastname: Iordanskiy
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Matthew
authors.identifier: http://orcid.org/0000-0002-9581-0377
authors.initials: M
authors.lastname: Berriman
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.
authors.forename: Michael I
authors.initials: MI
authors.lastname: Bukrinsky
authors.affiliation: Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.
authors.forename: Paul J
authors.identifier: http://orcid.org/0000-0003-1765-0002
authors.initials: PJ
authors.lastname: Brindley
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D049751:Genome, Helminth; D015658:HIV Infections; D015497:HIV-1; D051379:Mice; D016133:Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000821:virology*; D012555:Schistosomiasis mansoni / Q000821:virology*; D014161:Transduction, Genetic; D016662:Virus Integration