Background HIV-1 subtype B is the most prevalent in developed countries and, consequently, it has been extensively studied. On the other hand, subtype C is the most prevalent worldwide and therefore is a reasonable target for future studies. Here we evaluate the acquisition of resistance and the viability of HIV-1 subtype B and C RT clones from different isolates that were subjected to in vitro selection pressure with zidovudine (ZDV) and lamivudine (3TC). Methods/Principal Findings MT4 cells were infected with chimeric virus pseudotyped with RT from subtype B and C clones, which were previously subjected to serial passage with increasing concentrations of ZDV and 3TC. The samples collected after each passage were analyzed for the presence of resistance mutations and VL. No differences were found between subtypes B and C in viral load and resistance mutations when these viruses were selected with 3TC. However, the route of mutations and the time to rebound of subtype B and C virus were different when subjected to ZDV treatment. In order to confirm the role of the mutations detected, other clones were generated and subjected to in vitro selection. RT subtype B virus isolates tended to acquire different ZDV resistance mutations (Q151M and D67N or T215Y, D67D/N and F214L) compared to subtype C (D67N, K70R, T215I or T215F). Conclusions/Significance This study suggests that different subtypes have a tendency to react differently to antiretroviral drug selection in vitro. Consequently, the acquisition of resistance in patients undergoing antiretroviral therapy can be dependent on the subtypes composing the viral population.
References
[1]
Robertson DL, Anderson JP, Bradac JA, Carr JK, Funkhouser RK, et al. (2000) HIV-1 nomenclature proposal. Science 288: 55–56.
[2]
Wainberg MA (2004) HIV-1 subtype distribution and the problem of drug resistance. AIDS 18: S63–S68.
[3]
Perrin L, Telenti A (1998) HIV Treatment Failure: Testing for HIV Resistance in Clinical Practice. Science 280: 1871–1873.
[4]
Johnson VA, Calvez V, Günthard HF, Paredes R, Pillay D, et al. (2011) 2011 Update of the Drug Resistance Mutations in HIV-1. Top Ant Med 19: 156–164.
[5]
Marcelin AG, Delaugerre C, Wirden M, Viegas P, Simon A, et al. (2004) Thymidine Analogue Reverse Transcriptase Inhibitors Resistance Mutations Profiles and Association to Other Nucleoside Reverse Trasncriptase Inhibitors Resistance Mutations Observed in the Context of Virological Failure. J Med Virol 72: 162–165.
[6]
Renjifo B, Gilbert P, Chaplin B, Msamanga G, Mwakagile D, et al. (2004) Preferential in-utero transmission of HIV-1 subtype C as compared to HIV-1 subtype A or D. AIDS. 18: 1629–1636.
[7]
Gonzalez LM, Brindeiro RM, Tarin M, Calazams A, Soares MA, et al. (2003) In vitro hypersusceptibility of human immunodeficiency virus type 1 subtype C protease to Lopinavir. Antimicrob Agents Chemother 47: 2817–2822.
[8]
Carmona R, Perez-Alvarez L, Mu？oz M, Casado G, Delgade E, et al. (2005) Natural resistance-associated mutations to Enfuvirtide (T20) and polymorphisms in the gp41 region of different HIV-1 genetic forms from T20 naive patients. J Clin Virol 32: 248–253.
[9]
Dumans AT, Soares MA, Machado ES, Hué S, Brindeiro RM, et al. (2004) Synonymous genetic polymorphisms within Brazilian human immunodeficiency virus type 1 subtypes may influence mutational routes to drug resistance. J Infect Dis 189: 1232–1238.
[10]
Brenner BG, Oliveira M, Doualla-Bell F, Moisi DD, Ntemgwa M, et al. (2006) HIV-1 subtype C viruses rapidly develop K65R resistance to tenofovir in cell culture. AIDS 20: F9–F13.
[11]
Brindeiro RM, Diaz RS, Sabino EC, Morgado MG, Pires IL, et al. (2003) Brazilian Network for HIV Drug Resistance Surveillance (HIV-BResNet): a survey of chronically infected individuals. AIDS 17: 1063–1069.
[12]
Boucher CA, Keulen W, van Bommel T, Nijhuis M, de Jong D, et al. (1996) Human immunodeficiency virus type 1 drug susceptibility determination by using recombinant viruses generated from patient sera tested in a cell-killing assay. Antimicrob Agents Chemother 40: 2404–2409.
[13]
Aguiar RS, Pereira HS, Costa LJ, Brindeiro RM, Tanuri A (2006) Gag-Pol bearing a reverse transcriptase drug-resistant mutation influences viral genomic RNA incorporation into human immunodeficiency virus type 1 particles. J Gen Virol 87: 2669–2677.
[14]
Von Wyl V, Ehteshami M, Symons J, Bürgisser P, Nijhuis M, et al. (2010) Epidemiological and Biological Evidence for a Compensatory Effect of Connection Domain Mutation N348I on M184V in HIV-1 Reverse Transcriptase. J Infec Dis 201: 000–000.
[15]
van Maarseveen NM, Huigen MCDG, de Jong D, Smits AM, Boucher CAB, et al. (2006) A novel real-time PCR assay to determine relative replication capacity for HIV-1 protease variants and/or reverse transcriptase variants. J Virol Methods 133 (2): 185–194.
[16]
O’Doherty U, Swiggard WJ, Malim MH (2000) Human Immunodeficiency Virus Type 1 Spinoculation Enhances Infection through Virus Binding. J Virol 74: 10074–10080.
[17]
Ntemgwa ML, Toni T, Brenner BG, Oliveira M, Asahchop EL, et al. (2009) Nucleoside and Nucleotide Analogs Select in Culture for Different Patterns of Drug Resistance in Human Immunodeficiency Virus Types 1 and 2. Antimicrob Agents Chemother 53: 708–715.
[18]
Wainberg MA, Drosopoulos WC, Salomon H, Hsu M, Borkow G, et al. (1996) Enhanced fidelity of 3TC-selected mutant HIV-1 reverse transcriptase. Science 27: 1282–1285.
[19]
Rhee SY, Taylor J, Wadhera G, Ben-Hur A, Brutlag DL, et al. (2006) Genotypic predictors of human immunodeficiency virus type 1 drug resistance. Proc Natl Acad U S A 103: 17355–17360.
[20]
Svicher V, Sing T, Santoro MM, Forbici F, Rodríguez-Barrios F, et al. (2006) Involvement of Novel Human Immunodeficiency Virus Type 1 Reverse Transcriptase Mutations in the Regulation of Resistance to Nucleoside Inhibitors. J Virol 80: 7186–7198.
[21]
Melikian GL, Rhee SY, Taylor J, Fessel WJ, Kaufman D, et al. (2012) Standardized Comparison of the Relative Impacts of HIV-1 Reverse Transcriptase (RT) Mutations on Nucleoside RT Inhibitor Susceptibility. Antimicrob Agents Chemother 56: 2305–2313.
[22]
Rezende LF, Curr K, Ueno T, Mitsuva H, Prasad VR (1998) The impact of multidideoxynucleoside resistance-conferring mutations in human immunodeficiency virus type 1 reverse transcriptase on polymerase fidelity and error specificity. J Virol 72: 2890–2895.
[23]
Pellegin I, Izopet J, Reynes J, Denayrolles M, Montes B, et al. (1999) Emergence of zidovudine and multidrug-resistance mutations in the HIV-1 reverse transcriptase gene in therapy-naive patients receiving stavudine plus didanosine combination therapy. STADI Group. AIDS 13: 1705–1709.
[24]
Novitsky V, Wester CW, DeGruttola V, Bussmann H, Gaseitsiwe S, et al. (2007) The Reverse Transcriptase 67N 70R 215Y Genotype Is the Predominant TAM Pathway Associated with Virologic Failure among HIV Type 1C-Infected Adults Treated with ZDV/ddI-Containing HAART in Southern Africa. AIDS Res Hum Retroviruses 23: 868–878.
[25]
Armstrong KL, Lee TH, Essex M (2009) Replicative Capacity Differences of Thymidine Analog Resistance Mutations in Subtype B and C Human Immunodeficiency Virus Type 1. J Virol 83: 4051–4059.
[26]
Stürmer M, Staszewski S, Doerr HW, Larder B, Bloor S, et al. (2003) Correlation of Phenotypic Zidovudine Resistance with Mutational Patterns in the Reverse Transcriptase of Human Immunodeficiency Virus Type 1: Interpretation of established mutations and characterization of new polymorphisms at codons 208, 211, and 214. Antimicrob Agents Chemother. 47: 54–61.
[27]
Ceccherini-Silberstein F, Cozzi-Lepri A, Ruiz L, Mocroft A, Phillips AN, et al. (2007) Impact of HIV-1 Reverse Transcriptase Polymorphism F214L on Virological Response to Thymidine Analogue–Based Regimens in Antiretroviral Therapy (ART)–Naive and ART-Experienced Patients. J Infect Dis 196: 1180–90.
[28]
Puertas MC, Buzón M, Artese A, Alcaro S, Menedez-Arias L, et al. (2009) Effect of the Human Immunodeficiency Virus Type 1 Reverse Transcriptase Polymorphism Leu-214 on Replication Capacity and Drug Susceptibility. J Virol 83: 7434–7439.
[29]
Hu Z, Giguel F, Hatano H, Reid P, Lu J, et al. (2006) Fitness comparison of thymidine analog resistance pathways in human immunodeficiency virus type 1 J. Virol80(14): 7020–7.
[30]
Munerato P, Sucupira MC, Oliveros MP, Janini LM, de Souza DF, et al. (2010) HIV type 1 antiretroviral resistance mutations in subtypes B, C, and F in the City of S？o Paulo, Brazil. AIDS Res Hum Retroviruses 26: 265–73.
[31]
Marconi VC, Sunpath H, Lu Z, Gordon M, Koranteng-Apeagyei K, et al. (2008) Prevalence of HIV-1 drug resistance after failure of a first highly active antiretroviral therapy regimen in KwaZulu Natal, South Africa. Clin Infect Dis 46: 1589–97.
[32]
Santos AF, Soares MA (2010) HIV Genetic Diversity and Drug Resistance. Viruses 2: 503–31.
[33]
Kantor R, Katzenstein DA, Efron B, Carvalho AP, Wynhoven B, et al. (2005) Impact of HIV-1 subtype and antiretroviral therapy on protease and reverse transcriptase genotype: results of a global collaboration. PLoS Med 2: e112.
[34]
Novitsky V, Wester CW, DeGruttola V, Bussmann H, Gaseitsiwe S, et al. (2007) The reverse transcriptase 67N 70R 215Y genotype is the predominant TAM pathway associated with virologic failure among HIV type 1C-infected adults treated with ZDV/ddI-containing HAART in southern Africa. AIDS Res Hum Retroviruses 23: 868–78.
[35]
Rhee SY, Liu T, Ravela J, Gonzales MJ, Shafer RW (2004) Distribution of human immunodeficiency virus type 1 protease and reverse transcriptase mutation patterns in 4,183 persons undergoing genotypic resistance testing. Antimicrob Agents Chemother. 48: 3122–6.
[36]
Cozzi-Lepri A, Ruiz L, Loveday C, Phillips AN, Clotet B, et al. (2005) Thymidine analogue mutation profiles: factors associated with acquiring specific profiles and their impact on the virological response to therapy. Antivir Ther 10: 791–802.
[37]
Marcelin AG, Flandre P, Pavie J, Schmidely N, Wirden M, et al. (2005) Clinically relevant genotype interpretation of resistance to didanosine. Antimicrob Agents Chemother 49: 1739–44.
[38]
Miller MD, Margot N, Lu B, Zhong L, Chen SS, et al. (2004) Genotypic and phenotypic predictors of the magnitude of response to tenofovir disoproxil fumarate treatment in antiretroviral-experienced patients. J Infect Dis 189: 837–46.
