Isolation and characterization of mouse-human microcell hybrid cell clones permissive for infectious HIV particle release.
Mouse cells are non-permissive to human immunodeficiency virus type 1 (HIV) in that there is a pronounced post-integration block to viral replication. We have recently demonstrated that mouse-human somatic cell hybrids that contain human chromosome 2 increase both HIV Capsid (CA) production and infectious virus release. Here we report on the isolation of three mouse-human microcell hybrids (MCHs) that behave similarly, starting from a pool of 500 MCH clones. Release of virus was specific to HIV and cell revertants that no longer contained any human chromosome fragments did not release CA or infectious virus. Two of the three cell clones were identical as judged by PCR STS content and fluorescence in situ hybridization (FISH) and contained a single 2-12 human chromosome chimera. The third cell clone only contained human chromosome 12, as determined by PCR, FISH, and microarray analyses. There were no consistent differences in Gag protein and spliced/unspliced viral RNA levels between mouse cell lines. CMV promoter-driven, codon-optimized gag-pol had no effect on infectious HIV release from these mouse cells, despite allowing Gag targeting and increasing CA production. These permissive mouse-human MCHs and their corresponding non-permissive revertants may prove useful for mechanistic studies and also for identifying the responsible gene(s) or factor(s) involved in the production of HIV.
Coskun
Ayse K
AK
Baylor College of Medicine, Department of Molecular Virology and Microbiology, One Baylor Plaza, Houston, TX 77030, USA.
van Maanen
Marc
M
Janka
David
D
Stockton
David
D
Stankiewicz
Pawel
P
Stankiewicsz
Pawel
P
Yatsenko
Svetlana
S
Sutton
Richard E
RE
D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002881:Chromosomes, Human, Pair 12; D002889:Chromosomes, Human, Pair 2; D003062:Codon; D004279:DNA, Viral / Q000032:analysis; D015683:Gene Products, gag / Q000032:analysis / Q000096:biosynthesis; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D015497:HIV-1 / Q000254:growth & development*; D006801:Humans; D006822:Hybrid Cells / Q000821:virology*; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D046228:Microarray Analysis; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D054301:gag Gene Products, Human Immunodeficiency Virus
D003062:Codon; D004279:DNA, Viral; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D011498:Protein Precursors; D012367:RNA, Viral; D054301:gag Gene Products, Human Immunodeficiency Virus; C083868:gag protein p1, Human immunodeficiency virus; C059767:p55 gag precursor protein, Human immunodeficiency virus 1