The HR2 polymorphism N140I in the HIV-1 gp41 combined with the HR1 V38A mutation is associated with a less cytopathic phenotype

Retrovirology

Cecilia Cabrera³  Julià Blanco³  Bonaventura Clotet⁴  Carlo Federico Perno¹  Marta Curriu³  Nuria Pérez-Alvárez²  Valentina Svicher¹  Elisabet García³  Silvia Marfil³  Francesc Cunyat³ 

[1] Department of Experimental Medicine. University of "Tor Vergata," Rome, Italy;Statistics and Operation Research Department, Universitat Politècnica de Catalunya, Barcelona, Spain;IrsiCaixa-HIVACAT, Institut de Recerca en Ciències de la Salut Germans Trias i Pujol (IGTP), Hospital Germans Trias, Universitat Autònoma de Barcelona, Badalona 08916 Barcelona, Catalonia, Spain;Lluita contra la SIDA Foundation, Institut de Recerca en Ciències de la Salut Germans Trias i Pujol,Hospital Universitari Germans Trias i Pujol, Universitat Autònoma de Barcelona, 08916 Badalona, Barcelona, Spain

关键词: fusogenicity;    single cell death;    enfuvirtide;    gp41;    HIV;   

Others  :  1209338
DOI  :  10.1186/1742-4690-9-15

 received in 2011-11-09, accepted in 2012-02-14,  发布年份 2012

【 摘 要 】

Background

Resistance to the fusion inhibitor enfuvirtide (ENF) is achieved by changes in the gp41 subunit of the HIV envelope glycoprotein (Env). Specific ENF-associated mutational pathways correlate with immunological recovery, even after virological failure, suggesting that the acquisition of ENF resistance alters gp41 pathogenicity. To test this hypothesis, we have characterized the expression, fusion capability, induction of CD4⁺ T cell loss and single CD4⁺ T cell death of 48 gp41 proteins derived from three patients displaying different amino acids (N, T or I) at position 140 that developed a V38A mutation after ENF-based treatment.

Results

In all cases, intra-patient comparison of Env isolated pre- or post-treatment showed comparable values of expression and fusogenic capacity. Furthermore, Env with either N or T at position 140 induced comparable losses of CD4⁺ T-cells, irrespective of the residue present at position 38. Conversely, Env acquiring the V38A mutation in a 140I background induced a significantly reduced loss of CD4⁺ T cells and lower single-cell death than did their baseline controls. No altered ability to induce single-cell death was observed in the other clones.

Conclusions

Overall, primary gp41 proteins with both V38A and N140I changes showed a reduced ability to induce single cell death and deplete CD4⁺ T cells, despite maintaining fusion activity. The specificity of this phenotype highlights the relevance of the genetic context to the cytopathic capacity of Env and the role of ENF-resistance mutations in modulating viral pathogenicity in vivo, further supporting the hypothesis that gp41 is a critical mediator of HIV pathogenesis.

【 授权许可】

   
2012 Cunyat et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Hazenberg MD, Hamann D, Schuitemaker H, Miedema F: T cell depletion in HIV-1 infection: how CD4+ T cells go out of stock. Nat Immunol 2000, 1(4):285-9.
• [2]McCune JM: The dynamics of CD4+ T-cell depletion in HIV disease. Nature 2001, 410(6831):974-9.
• [3]Finkel TH, Banda NK: Indirect mechanisms of HIV pathogenesis: how does HIV kill T cells? Curr Opin Immunol 1994, 6(4):605-15.
• [4]Gandhi RT, Chen BK, Straus SE, Dale JK, Lenardo MJ, Baltimore D: HIV-1 directly kills CD4+ T cells by a Fas-independent mechanism. J Exp Med 1998, 187(7):1113-22.
• [5]Douek DC, Roederer M, Koup RA: Emerging concepts in the immunopathogenesis of AIDS. Annu Rev Med 2009, 60:471-84.
• [6]Ameisen JC, Capron A: Cell dysfunction and depletion in AIDS: the programmed cell death hypothesis. Immunol Today 1991, 12(4):102-5.
• [7]Gougeon ML: Apoptosis as an HIV strategy to escape immune attack. Nat Rev Immunol 2003, 3(5):392-404.
• [8]Perfettini JL, Castedo M, Roumier T, Andreau K, Nardacci R, Piacentini M, Kroemer G: Mechanisms of apoptosis induction by the HIV-1 envelope. Cell Death Differ 2005, 12(Suppl 1):916-23.
• [9]Badley AD, Pilon AA, Landay A, Lynch DH: Mechanisms of HIV-associated lymphocyte apoptosis. Blood 2000, 96(9):2951-64.
• [10]Gougeon ML, Montagnier L: Apoptosis in AIDS. Science 1993, 260(5112):1269-70.
• [11]Doitsh G, Cavrois M, Lassen KG, Zepeda O, Yang Z, Santiago ML, Hebbeler AM, Greene WC: Abortive HIV infection mediates CD4 T cell depletion and inflammation in human lymphoid tissue. Cell 2010, 143(5):789-801.
• [12]Doms RW, Moore JP: HIV-1 membrane fusion: targets of opportunity. J Cell Biol 2000, 151(2):F9-14.
• [13]Doms RW, Trono D: The plasma membrane as a combat zone in the HIV battlefield. Genes Dev 2000, 14(21):2677-88.
• [14]Eckert DM, Kim PS: Mechanisms of viral membrane fusion and its inhibition. Annu Rev Biochem 2001, 70:777-810.
• [15]Ferri KF, Jacotot E, Blanco J, Este JA, Zamzami N, Susin SA, Xie Z, Brothers G, Reed JC, Penninger JM, Kroemer G: Apoptosis control in syncytia induced by the HIV type 1-envelope glycoprotein complex: role of mitochondria and caspases. J Exp Med 2000, 192(8):1081-92.
• [16]Ahr B, Robert-Hebmann V, Devaux C, Biard-Piechaczyk M: Apoptosis of uninfected cells induced by HIV envelope glycoproteins. Retrovirology 2004, 1:12. BioMed Central Full Text
• [17]Moore JP, McKeating JA, Weiss RA, Sattentau QJ: Dissociation of gp120 from HIV-1 virions induced by soluble CD4. Science 1990, 250(4984):1139-42.
• [18]Willey RL, Martin MA: Association of human immunodeficiency virus type 1 envelope glycoprotein with particles depends on interactions between the third variable and conserved regions of gp120. J Virol 1993, 67(6):3639-43.
• [19]Wyatt R, Sodroski J: The HIV-1 envelope glycoproteins: fusogens, antigens, and immunogens. Science 1998, 280(5371):1884-8.
• [20]Roux KH, Taylor KA: AIDS virus envelope spike structure. Curr Opin Struct Biol 2007, 17(2):244-52.
• [21]Wild C, Dubay JW, Greenwell T, Baird T Jr, Oas TG, McDanal C, Hunter E, Matthews T: Propensity for a leucine zipper-like domain of human immunodeficiency virus type 1 gp41 to form oligomers correlates with a role in virus-induced fusion rather than assembly of the glycoprotein complex. Proc Natl Acad Sci USA 1994, 91(26):12676-80.
• [22]Wild CT, Shugars DC, Greenwell TK, McDanal CB, Matthews TJ: Peptides corresponding to a predictive alpha-helical domain of human immunodeficiency virus type 1 gp41 are potent inhibitors of virus infection. Proc Natl Acad Sci USA 1994, 91(21):9770-4.
• [23]Kilgore NR, Salzwedel K, Reddick M, Allaway GP, Wild CT: Direct evidence that C-peptide inhibitors of human immunodeficiency virus type 1 entry bind to the gp41 N-helical domain in receptor-activated viral envelope. J Virol 2003, 77(13):7669-72.
• [24]Cervia JS, Smith MA: Enfuvirtide (T-20): a novel human immunodeficiency virus type 1 fusion inhibitor. Clin Infect Dis 2003, 37(8):1102-6.
• [25]Kilby JM, Hopkins S, Venetta TM, DiMassimo B, Cloud GA, Lee JY, Alldredge L, Hunter E, Lambert D, Bolognesi D, Matthews T, Johnson MR, Nowak MA, Shaw GM, Saag MS: Potent suppression of HIV-1 replication in humans by T-20, a peptide inhibitor of gp41-mediated virus entry. Nat Med 1998, 4(11):1302-7.
• [26]Lazzarin A, Clotet B, Cooper D, Reynes J, Arasteh K, Nelson M, Katlama C, Stellbrink HJ, Delfraissy JF, Lange J, Huson L, DeMasi R, Wat C, Delehanty J, Drobnes C, Salgo M: Efficacy of enfuvirtide in patients infected with drug-resistant HIV-1 in Europe and Australia. N Engl J Med 2003, 348(22):2186-95.
• [27]Lalezari JP, Henry K, O'Hearn M, Montaner JS, Piliero PJ, Trottier B, Walmsley S, Cohen C, Kuritzkes DR, Eron JJ Jr, Chung J, DeMasi R, Donatacci L, Drobnes C, Delehanty J, Salgo M: Enfuvirtide, an HIV-1 fusion inhibitor, for drug-resistant HIV infection in North and South America. N Engl J Med 2003, 348(22):2175-85.
• [28]Bonjoch A, Negredo E, Puig J, Erkizia I, Puig T, Cabrera C, Ruiz L, Clotet B: Viral failure in HIV-infected patients with long-lasting viral suppression who discontinued enfuvirtide. AIDS 2006, 20(14):1896-8.
• [29]Cabrera C, Marfil S, Garcia E, Martinez-Picado J, Bonjoch A, Bofill M, Moreno S, Ribera E, Domingo P, Clotet B, Ruiz L: Genetic evolution of gp41 reveals a highly exclusive relationship between codons 36, 38 and 43 in gp41 under long-term enfuvirtide-containing salvage regimen. AIDS 2006, 20(16):2075-80.
• [30]Wei X, Decker JM, Liu H, Zhang Z, Arani RB, Kilby JM, Saag MS, Wu X, Shaw GM, Kappes JC: Emergence of resistant human immunodeficiency virus type 1 in patients receiving fusion inhibitor (T-20) monotherapy. Antimicrob Agents Chemother 2002, 46(6):1896-905.
• [31]Lu J, Deeks SG, Hoh R, Beatty G, Kuritzkes BA, Martin JN, Kuritzkes DR: Rapid emergence of enfuvirtide resistance in HIV-1-infected patients: results of a clonal analysis. J Acquir Immune Defic Syndr 2006, 43(1):60-4.
• [32]Ray N, Blackburn LA, Doms RW: HR-2 mutations in human immunodeficiency virus type 1 gp41 restore fusion kinetics delayed by HR-1 mutations that cause clinical resistance to enfuvirtide. J Virol 2009, 83(7):2989-95.
• [33]Reeves JD, Gallo SA, Ahmad N, Miamidian JL, Harvey PE, Sharron M, Pohlmann S, Sfakianos JN, Derdeyn CA, Blumenthal R, Hunter E, Doms RW: Sensitivity of HIV-1 to entry inhibitors correlates with envelope/coreceptor affinity, receptor density, and fusion kinetics. Proc Natl Acad Sci USA 2002, 99(25):16249-54.
• [34]Tolstrup M, Selzer-Plon J, Laursen AL, Bertelsen L, Gerstoft J, Duch M, Pedersen FS, Ostergaard L: Full fusion competence rescue of the enfuvirtide resistant HIV-1 gp41 genotype (43D) by a prevalent polymorphism (137K). AIDS 2007, 21(4):519-21.
• [35]Xu L, Pozniak A, Wildfire A, Stanfield-Oakley SA, Mosier SM, Ratcliffe D, Workman J, Joall A, Myers R, Smit E, Cane PA, Greenberg ML, Pillay D: Emergence and evolution of enfuvirtide resistance following long-term therapy involves heptad repeat 2 mutations within gp41. Antimicrob Agents Chemother 2005, 49(3):1113-9.
• [36]Jenwitheesuk E, Samudrala R: Heptad-repeat-2 mutations enhance the stability of the enfuvirtide-resistant HIV-1 gp41 hairpin structure. Antivir Ther 2005, 10(8):893-900.
• [37]Baatz F, Nijhuis M, Lemaire M, Riedijk M, Wensing AM, Servais JY, van Ham PM, Hoepelman AI, Koopmans PP, Sprenger HG, Devaux C, Schmit JC, Perez Bercoff D: Impact of the HIV-1 env genetic context outside HR1-HR2 on resistance to the fusion inhibitor enfuvirtide and viral infectivity in clinical isolates. PLoS One 2011, 6(7):e21535.
• [38]Svicher V, Aquaro S, D'Arrigo R, Artese A, Dimonte S, Alcaro S, Santoro MM, Di Perri G, Caputo SL, Bellagamba R, Zaccarelli M, Visco-Comandini U, Antinori A, Narciso P, Ceccherini-Silberstein F, Perno CF: Specific enfuvirtide-associated mutational pathways in HIV-1 Gp41 are significantly correlated with an increase in CD4(+) cell count, despite virological failure. J Infect Dis 2008, 197(10):1408-18.
• [39]Aquaro S, D'Arrigo R, Svicher V, Perri GD, Caputo SL, Visco-Comandini U, Santoro M, Bertoli A, Mazzotta F, Bonora S, Tozzi V, Bellagamba R, Zaccarelli M, Narciso P, Antinori A, Perno CF: Specific mutations in HIV-1 gp41 are associated with immunological success in HIV-1-infected patients receiving enfuvirtide treatment. J Antimicrob Chemother 2006, 58(4):714-22.
• [40]Melby TE, Despirito M, Demasi RA, Heilek G, Thommes JA, Greenberg ML, Graham N: Association between specific enfuvirtide resistance mutations and CD4 cell response during enfuvirtide-based therapy. AIDS 2007, 21(18):2537-9.
• [41]Blanco J, Barretina J, Ferri KF, Jacotot E, Gutierrez A, Armand-Ugon M, Cabrera C, Kroemer G, Clotet B, Este JA: Cell-surface-expressed HIV-1 envelope induces the death of CD4 T cells during GP41-mediated hemifusion-like events. Virology 2003, 305(2):318-29.
• [42]Garg H, Joshi A, Freed EO, Blumenthal R: Site-specific mutations in HIV-1 gp41 reveal a correlation between HIV-1-mediated bystander apoptosis and fusion/hemifusion. J Biol Chem 2007, 282(23):16899-906.
• [43]Garg H, Joshi A, Blumenthal R: Altered bystander apoptosis induction and pathogenesis of enfuvirtide-resistant HIV type 1 Env mutants. AIDS Res Hum Retroviruses 2009, 25(8):811-7.
• [44]Garg H, Joshi A, Ye C, Shankar P, Manjunath N: Single amino acid change in gp41 region of HIV-1 alters bystander apoptosis and CD4 decline in humanized mice. Virol J 2011, 8:34. BioMed Central Full Text
• [45]Goubard A, Clavel F, Mammano F, Labrosse B: In vivo selection by enfuvirtide of HIV type-1 env quasispecies with optimal potential for phenotypic expression of HR1 mutations. Antivir Ther 2009, 14(4):597-602.
• [46]Labrosse B, Morand-Joubert L, Goubard A, Rochas S, Labernardiere JL, Pacanowski J, Meynard JL, Hance AJ, Clavel F, Mammano F: Role of the envelope genetic context in the development of enfuvirtide resistance in human immunodeficiency virus type 1-infected patients. J Virol 2006, 80(17):8807-19.
• [47]Wang XM, Nadeau PE, Lo YT, Mergia A: Caveolin-1 modulates HIV-1 envelope-induced bystander apoptosis through gp41. J Virol 2010, 84(13):6515-26.
• [48]Tersmette M, Lange JM, de Goede RE, de Wolf F, Eeftink-Schattenkerk JK, Schellekens PT, Coutinho RA, Huisman JG, Goudsmit J, Miedema F: Association between biological properties of human immunodeficiency virus variants and risk for AIDS and AIDS mortality. Lancet 1989, 1(8645):983-5.
• [49]Blanco J, Barretina J, Clotet B, Este JA: R5 HIV gp120-mediated cellular contacts induce the death of single CCR5-expressing CD4 T cells by a gp41-dependent mechanism. J Leukoc Biol 2004, 76(4):804-11.
• [50]Sterjovski J, Churchill MJ, Ellett A, Gray LR, Roche MJ, Dunfee RL, Purcell DF, Saksena N, Wang B, Sonza S, Wesselingh SL, Karlsson I, Fenyo EM, Gabuzda D, Cunningham AL, Gorry PR: Asn 362 in gp120 contributes to enhanced fusogenicity by CCR5-restricted HIV-1 envelope glycoprotein variants from patients with AIDS. Retrovirology 2007, 4:89. BioMed Central Full Text
• [51]Karlsson GB, Halloran M, Schenten D, Lee J, Racz P, Tenner-Racz K, Manola J, Gelman R, Etemad-Moghadam B, Desjardins E, Wyatt R, Gerard NP, Marcon L, Margolin D, Fanton J, Axthelm MK, Letvin NL, Sodroski J: The envelope glycoprotein ectodomains determine the efficiency of CD4+ T lymphocyte depletion in simian-human immunodeficiency virus-infected macaques. J Exp Med 1998, 188(6):1159-71.
• [52]Etemad-Moghadam B, Sun Y, Nicholson EK, Fernandes M, Liou K, Gomila R, Lee J, Sodroski J: Envelope glycoprotein determinants of increased fusogenicity in a pathogenic simian-human immunodeficiency virus (SHIV-KB9) passaged in vivo. J Virol 2000, 74(9):4433-40.
• [53]Etemad-Moghadam B, Rhone D, Steenbeke T, Sun Y, Manola J, Gelman R, Fanton JW, Racz P, Tenner-Racz K, Axthelm MK, Letvin NL, Sodroski J: Membrane-fusing capacity of the human immunodeficiency virus envelope proteins determines the efficiency of CD+ T-cell depletion in macaques infected by a simian-human immunodeficiency virus. J Virol 2001, 75(12):5646-55.
• [54]LaBonte JA, Patel T, Hofmann W, Sodroski J: Importance of membrane fusion mediated by human immunodeficiency virus envelope glycoproteins for lysis of primary CD4-positive T cells. J Virol 2000, 74(22):10690-8.
• [55]Chen SS, Yang P, Ke PY, Li HF, Chan WE, Chang DK, Chuang CK, Tsai Y, Huang SC: Identification of the LWYIK motif located in the human immunodeficiency virus type 1 transmembrane gp41 protein as a distinct determinant for viral infection. J Virol 2009, 83(2):870-83.
• [56]Gougeon ML, Lecoeur H, Dulioust A, Enouf MG, Crouvoiser M, Goujard C, Debord T, Montagnier L: Programmed cell death in peripheral lymphocytes from HIV-infected persons: increased susceptibility to apoptosis of CD4 and CD8 T cells correlates with lymphocyte activation and with disease progression. J Immunol 1996, 156(9):3509-20.
• [57]Debatin KM, Fahrig-Faissner A, Enenkel-Stoodt S, Kreuz W, Benner A, Krammer PH: High expression of APO-1 (CD95) on T lymphocytes from human immunodeficiency virus-1-infected children. Blood 1994, 83(10):3101-3.
• [58]Finkel TH, Tudor-Williams G, Banda NK, Cotton MF, Curiel T, Monks C, Baba TW, Ruprecht RM, Kupfer A: Apoptosis occurs predominantly in bystander cells and not in productively infected cells of HIV- and SIV-infected lymph nodes. Nat Med 1995, 1(2):129-34.
• [59]De Clercq E, Yamamoto N, Pauwels R, Balzarini J, Witvrouw M, De Vreese K, Debyser Z, Rosenwirth B, Peichl P, Datema R, Thornton D, Skerlj R, Gaul F, Padmanabhan S, Bridger G, Henson G, Abrams M: Highly potent and selective inhibition of human immunodeficiency virus by the bicyclam derivative JM3100. Antimicrob Agents Chemother 1994, 38(4):668-74.
• [60]Baba M, Nishimura O, Kanzaki N, Okamoto M, Sawada H, Iizawa Y, Shiraishi M, Aramaki Y, Okonogi K, Ogawa Y, Meguro K, Fujino M: A small-molecule, nonpeptide CCR5 antagonist with highly potent and selective anti-HIV-1 activity. Proc Natl Acad Sci USA 1999, 96(10):5698-703.
• [61]Dragic T, Trkola A, Thompson DA, Cormier EG, Kajumo FA, Maxwell E, Lin SW, Ying W, Smith SO, Sakmar TP, Moore JP: A binding pocket for a small molecule inhibitor of HIV-1 entry within the transmembrane helices of CCR5. Proc Natl Acad Sci USA 2000, 97(10):5639-44.
• [62]Malim MH, Hauber J, Fenrick R, Cullen BR: Immunodeficiency virus rev trans-activator modulates the expression of the viral regulatory genes. Nature 1988, 335(6186):181-3.
• [63]Li M, Gao F, Mascola JR, Stamatatos L, Polonis VR, Koutsoukos M, Voss G, Goepfert P, Gilbert P, Greene KM, Bilska M, Kothe DL, Salazar-Gonzalez JF, Wei X, Decker JM, Hahn BH, Montefiori DC: Human immunodeficiency virus type 1 env clones from acute and early subtype B infections for standardized assessments of vaccine-elicited neutralizing antibodies. J Virol 2005, 79(16):10108-25.