title: Survival of the fittest: positive selection of CD4+ T cells expressing a membrane-bound fusion inhibitor following HIV-1 infection.
abstract: Although a variety of genetic strategies have been developed to inhibit HIV replication, few direct comparisons of the efficacy of these inhibitors have been carried out. Moreover, most studies have not examined whether genetic inhibitors are able to induce a survival advantage that results in an expansion of genetically-modified cells following HIV infection. We evaluated the efficacy of three leading genetic strategies to inhibit HIV replication: 1) an HIV-1 tat/rev-specific small hairpin (sh) RNA; 2) an RNA antisense gene specific for the HIV-1 envelope; and 3) a viral entry inhibitor, maC46. In stably transduced cell lines selected such that >95% of cells expressed the genetic inhibitor, the RNA antisense envelope and viral entry inhibitor maC46 provided the strongest inhibition of HIV-1 replication. However, when mixed populations of transduced and untransduced cells were challenged with HIV-1, the maC46 fusion inhibitor resulted in highly efficient positive selection of transduced cells, an effect that was evident even in mixed populations containing as few as 1% maC46-expressing cells. The selective advantage of the maC46 fusion inhibitor was also observed in HIV-1-infected cultures of primary T lymphocytes as well as in HIV-1-infected humanized mice. These results demonstrate robust inhibition of HIV replication with the fusion inhibitor maC46 and the antisense Env inhibitor, and importantly, a survival advantage of cells expressing the maC46 fusion inhibitor both in vitro and in vivo. Evaluation of the ability of genetic inhibitors of HIV-1 replication to confer a survival advantage on genetically-modified cells provides unique information not provided by standard techniques that may be important in the in vivo efficacy of these genes.
authors.affiliation: Angewandte Virologie und Gentherapie, Chemotherapeutisches Forschungsinstitut Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Kimpel
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Braun
authors.forename: Gang
authors.initials: G
authors.lastname: Qiu
authors.forename: Fay Eng
authors.initials: FE
authors.lastname: Wong
authors.forename: Michelle
authors.initials: M
authors.lastname: Conolle
authors.forename: Jörn E
authors.initials: JE
authors.lastname: Schmitz
authors.forename: Christian
authors.initials: C
authors.lastname: Brendel
authors.forename: Laurent M
authors.initials: LM
authors.lastname: Humeau
authors.forename: Boro
authors.initials: B
authors.lastname: Dropulic
authors.forename: John J
authors.initials: JJ
authors.lastname: Rossi
authors.forename: Annemarie
authors.initials: A
authors.lastname: Berger
authors.forename: Dorothee
authors.initials: D
authors.lastname: von Laer
authors.forename: R Paul
authors.initials: RP
authors.lastname: Johnson
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D015870:Gene Expression; D015316:Genetic Therapy; D023581:HIV Fusion Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D015658:HIV Infections / Q000235:genetics / Q000628:therapy; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D051379:Mice; D016372:RNA, Antisense / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D013045:Species Specificity; D014161:Transduction, Genetic; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: C515020:C46 HIV-1 fusion inhibitory peptide; D023581:HIV Fusion Inhibitors; D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins
references.citation: J Gene Med. 2004 Sep;6(9):963-73
references.pmid: 15352069
references.citation: Proc Natl Acad Sci U S A. 2007 Aug 7;104(32):13110-5
references.pmid: 17670939
references.citation: Mol Ther. 2010 Apr;18(4):803-11
references.pmid: 20104212
references.citation: Nat Biotechnol. 2008 Jul;26(7):808-16
references.pmid: 18587387
references.citation: Nat Genet. 1997 May;16(1):100-3
references.pmid: 9140404
references.citation: Mol Ther. 2004 Jun;9(6):902-13
references.pmid: 15194057
references.citation: Semin Immunol. 2008 Jun;20(3):181-6
references.pmid: 18595731
references.citation: J Virol. 2005 Aug;79(16):10237-46
references.pmid: 16051817
references.citation: Hum Gene Ther. 2006 Jun;17(6):577-88
references.pmid: 16776567
references.citation: J Virol. 2004 Jul;78(13):7079-88
references.pmid: 15194784
references.citation: J Gene Med. 2005 May;7(5):552-64
references.pmid: 15655805
references.citation: N Engl J Med. 2009 Feb 12;360(7):692-8
references.pmid: 19213682
references.citation: Clin Microbiol Infect. 2009 Jan;15 Suppl 1:69-73
references.pmid: 19220361
references.citation: J Med Primatol. 1992 Jul;21(5):265-9
references.pmid: 1383547
references.citation: Front Biosci. 2006 Jan 01;11:838-51
references.pmid: 16146775
references.citation: Mol Ther. 2005 Jul;12(1):77-86
references.pmid: 15963923
references.citation: Lancet. 2005 Jul 30-Aug 5;366(9483):378-84
references.pmid: 16054937
references.citation: Mol Ther. 2005 Dec;12(6):1157-67
references.pmid: 16168713
references.citation: CMAJ. 2004 Jan 20;170(2):229-38
references.pmid: 14734438
references.citation: J Theor Biol. 2006 Jan 7;238(1):60-77
references.pmid: 15993899
references.citation: Sci Transl Med. 2010 Jun 16;2(36):36ra43
references.pmid: 20555022
references.citation: Gene Ther. 2006 Oct;13(20):1480-92
references.pmid: 16738691
references.citation: J Virol. 2001 Mar;75(6):3038-42
references.pmid: 11222732
references.citation: Hum Gene Ther. 2009 Apr;20(4):325-36
references.pmid: 19267665
references.citation: Nat Biotechnol. 2002 May;20(5):500-5
references.pmid: 11981565
references.citation: J Virol. 2002 Feb;76(3):1488-95
references.pmid: 11773421
references.citation: Mol Ther. 2007 May;15(5):1024-33
references.pmid: 17356541
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17372-7
references.pmid: 17090675
references.citation: Hum Gene Ther. 2004 Mar;15(3):251-62
references.pmid: 15018734
references.citation: Nat Biotechnol. 1997 Sep;15(9):871-5
references.pmid: 9306402
references.citation: Mol Ther. 2005 Nov;12(5):900-9
references.pmid: 16115802
references.citation: Hum Gene Ther. 2005 Sep;16(9):1065-74
references.pmid: 16149905
references.citation: J Virol. 2004 Jan;78(2):568-75
references.pmid: 14694088
references.citation: Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2889-94
references.pmid: 8610137
references.citation: Hum Gene Ther. 2002 May 1;13(7):803-13
references.pmid: 11975847
references.citation: J Postgrad Med. 2006 Jul-Sep;52(3):197-200
references.pmid: 16855321
references.citation: Gene Ther. 2008 Sep;15(17):1210-22
references.pmid: 18449216
references.citation: Nat Biotechnol. 2007 Dec;25(12):1444-54
references.pmid: 18066041