RESULTS Qualitative analysis of HLA class I A2-, A3-, and B7-restricted CTL responses in HIV-1-infected individuals by bulk stimulation of PBMC. To begin to address the relative contribution of single HLA class I alleles to the overall CTL response, we recruited three persons matched at the HLA A2, A3, and B7 alleles. CTL responses to 12 optimal A2-, A3-, and B7-restricted epitopes known at this time were analyzed by bulk stimulation of PBMC. These and subsequent epitopes chosen for use in this study (Table 1) contain motifs important for binding the relevant class I HLA allele ( 20) and were defined as optimal epitopes using truncated peptides at limiting concentrations ( 9). All epitopes were shown to be processed endogenously in infected cells, using vaccinia virus recombinants to express antigen and CTL clones specific for the epitopes in Table 1 (data not shown). PBMC were pulsed with the A2, A3, or B7 peptides indicated, expanded for 10 days, and then assayed for the ability to lyse autologous B-LCL pulsed with the relevant peptides. Significant heterogeneity in the dominance and breadth of responses was demonstrated to these epitopes in the three individuals analyzed (Fig. 1). The dominant A2-restricted response in subject 161j is clearly directed against the Gag epitope p17 77–85, whereas no clearly dominant A2-restricted response could be identified in subjects 13070 and 221L (Fig. 1A). For the A3-restricted epitopes (Fig. 1B), subject 161j targeted two codominant epitopes (p17 18–26 and gp41 770–780) in addition to two other subdominant responses (p17 20–28 and Nef 73–82). In contrast, the dominant epitope for subject 13070 (Nef 73–82) was recognized least in subject 161j. The strongest A3-restricted response in subject 221L was directed against the p17 18–26 epitope, which was similarly targeted as a dominant epitope by subject 161j. Of the B7-restricted epitopes tested, the dominant responses in subject 161j were directed against gp41 843–851 and Nef 128–137. The most dominant epitope targeted by both 13070 and 221L was Nef 128–137 (Fig. 1C). These results indicate significant heterogeneity in the dominance of responses in persons of the same HLA type, as measured following in vitro stimulation of PBMC. | FIG. 1HLA-A2-, A3-, and B7-restricted HIV-1-specific CTL responses in 161j, 13070, and 221L as determined by bulk stimulation of PBMC, followed by chromium release assay using B-LCL pulsed with the indicated peptides and nonpulsed B-LCL. (A) CTL responses to (more ...) |
Breadth and magnitude of CTL responses by Elispot assay to optimal HIV-1 epitopes restricted by HLA class I A and B alleles in HIV-1-infected individuals with shared HLA class I alleles. The above assays rely on in vitro expansion of PBMC and require large numbers of cells. Newer methods assessing cytokine production by antigen-specific CD8 cells allow for rapid, direct quantitation and require fewer cells. We therefore expanded our analysis to include five additional HLA A2-, A3-, and B7-positive HIV-1-infected individuals, using the IFN-γ Elispot assay to evaluate CTL responses to 27 optimal A2-, A3-, or B7-restricted epitopes in all eight subjects. Subjects 11324, 11841, 13070, 16732, 221L, and 161j all have chronic HIV-1 infection. AC-03 and OP337 were both identified with acute HIV-1 infection. All subjects except for 161j were receiving antiretroviral therapy, although virus was still detectable (>50 RNA copies/ml of plasma) in four subjects. CTL responses to eight optimal A2-restricted epitopes were evaluated in each subject. p17 77–85 was the dominant epitope recognized in all subjects with chronic HIV-1 infection, except for subject 11841 for which RT 33–41 appeared to be the dominant A2-restricted epitope targeted. Subject 11841 also had the broadest A2-restricted HIV-1-specific CTL response, with five of the eight A2-restricted epitopes tested being targeted. Three of the subjects (11324, 16732, and 221L) had detectable A2-restricted CTL responses directed against the p17 77–85 epitope only. Subjects 13070 and 161j had very weak responses to the A2-restricted RT 33–41 and RT 309–317 epitopes, respectively, in addition to the p17 77–85 epitope. In contrast, both subjects with acute HIV-1 infection failed to recognize the p17 77–85 epitope. No A2-restricted epitopes were detected in AC-03, whereas OP337 recognized only the gp41 813–822 epitope. The dominance of the p17 77–85 epitope in subjects with chronic HIV-1 infection and the lack of response in individuals with acute HIV-1 infection are consistent with previously published studies of A2-restricted responses ( 10, 21, 24, 26, 42, 57). Furthermore, the magnitude of responses to each epitope was highly variable among individuals. The sum total number of A2-restricted CTL responses differed over 40-fold, ranging from 70 SFC/million PBMC in subject 16732 to 2,820 SFC/million PBMC in subject 161j. CTL responses to eight optimally defined A3-restricted HIV-1 epitopes were similarly evaluated in these same eight individuals (Fig. 2). All subjects recognized at least one A3-restricted epitope (range, one to eight), but there was no clearly dominant epitope targeted by all subjects as had been observed with the A2-restricted epitope p17 77–85. In subjects 11324 and 11841, RT 158–166 is clearly targeted as the dominant A3-restricted response. The Elispot results indicate that in subjects 13070 and 16732, Nef 73–82 was the dominant A3-restricted epitope targeted. However, three A3-restricted epitopes were recognized in 221L with no one dominant epitope clearly targeted. All eight A3-restricted epitopes targeted were recognized by 161j, with epitopes p17 18–26 and gp41 770–780 generating the strongest CTL responses in the Elispot assay. Furthermore, there was over a 100-fold difference in the overall total magnitude of CTL responses to A3-restricted epitopes among the subjects evaluated, ranging from 40 SFC/million PBMC in subject 16732 to 4,700 SFC/million PBMC in subject 161j. | FIG. 2HIV-1-specific CTL responses in eight HIV-1-infected individuals as determined by IFN-γ Elispot assay. All optimal HIV-1 epitopes defined (9) were tested for each individual's HLA class I A and B alleles. Only epitopes recognized with more than (more ...) |
Eleven previously defined B7-restricted optimal epitopes were also tested in the Elispot assay in the eight subjects included in this study. As shown in Fig. 2, all subjects with chronic HIV-1 infection recognized at least two B7-restricted epitopes (range, two to eight). Subject OP337, who is homozygous for the B7 allele, recognized five B7-restricted epitopes. However, the other subject with acute HIV-1 infection, AC-03, failed to recognize any of the 11 B7-restricted epitopes tested. Similar to the results seen with the A3-restricted epitopes, the B7-restricted CTL response was highly variable among all subjects and there was no clearly dominant epitope targeted in most individuals. Figure 3 summarizes the breadth of CTL responses to all the optimal A2-, A3-, and B7-restricted CTL responses in the eight subjects analyzed. | FIG. 3Frequency of recognition of all optimal A2-, A3-, and B7-restricted HIV-1 epitopes among the eight HIV-1-infected individuals included in this study. Gray boxes represent a positive CTL response in a subject by Elispot to a given epitope. Responses greater (more ...) |
Relative contributions of CTL responses restricted by the second B allele in persons coexpressing A2, A3, and B7. The above results included analyses of the CTL response in each subject to HIV-1 epitopes restricted by the three HLA class I alleles shared by all of the subjects. We also determined CTL responses to optimal epitopes defined for each subject's HLA class I unmatched B allele in order to gain a more comprehensive view of the total HIV-1-specific CTL response in each individual. These alleles, including B8, B27, B44, B60, B61, and B62, are less common in the population, and there are fewer optimally defined epitopes for these alleles than for the A2, A3, and B7 alleles ( 9). As shown in Fig. 2, no immunodominant epitopes were recognized in subjects 13070 and 11324 in response to the five B61- and four B62-restricted epitopes tested, respectively. However, in subject 221L, the B8-restricted epitope Nef 90-97 was the second highest response in magnitude compared to all the HLA class I A- and B-restricted epitopes tested in this individual. Moreover, B8 accounted for about one-third of the total CTL response. Five B60-restricted epitopes were tested in subject 161j, and responses were detected against all five epitopes. The strongest CTL response detected in subject 161j was directed against the B60-restricted epitope p24 44–52, and all the B60-restricted responses together contributed over one-third of the total CTL response. Overall, the unmatched HLA class I B allele contributed between 0 and 38% of the total magnitude of the response, and the numbers of epitopes targeted through this fourth allele ranged from 0 to 5. These studies provide additional quantitative evidence that the magnitude and breadth of CTL responses differ considerably in persons of similar HLA types and indicate that the assessment of responses to three alleles still significantly underestimates the breadth and magnitude of the HIV-1-specific CTL response. The breadth and magnitude of the HIV-1-specific CTL responses in the above experiments were determined by IFN-γ production in an Elispot assay. In order to obtain a more precise definition of both the phenotype and the quantity of responding cells, we performed a more comprehensive analysis of the HIV-1-specific CTL responses in subject 161j by flow cytometry-based ICS assays. Thirty-two A2-, A3-, B7-, or B60-restricted epitopes were tested by Elispot assay in subject 161j, of which 22 epitopes generated positive responses (Fig. 2). Eighteen epitopes generated responses with a magnitude of >100 SFC/million PBMC. These 18 epitopes were then tested in an ICS assay for IFN-γ production. Representative ICS data for 10 immunogenic epitopes recognized by subject 161j are indicated in Fig. 4. Of the 18 epitopes tested by ICS, the sum total percentage of CD8 + T lymphocytes specific for HIV-1 in subject 161j was 10.9% (Fig. 4 and data not shown). These data confirm the magnitude and breadth of CTL responses and together with the Elispot data provide firm evidence that the immune response can be extremely broadly directed in some persons. | FIG. 4Recognition of optimally defined A2-, A3-, B7-, and B60-restricted HIV-1 epitopes in subject 161j by ICS for IFN-γ production. The percentage of CD8+ T cells expressing IFN-γ (minus background IFN-γ production) is indicated (more ...) |
Relative contributions of HLA class I A- and B-restricted epitopes to the overall HIV-1-specific CTL immune response. The above studies indicate heterogeneity in the magnitude and breadth of CTL responses to individual known HIV-1 epitopes in persons sharing multiple MHC class I alleles. We next analyzed the relative contributions of the HLA class I A and B alleles to the total HIV-1-specific CTL response in the eight individuals included in this study. The sum of the CTL responses as determined by Elispot assay for each HLA class I A and B allele are shown in Fig. 5 for each subject. The overall highest magnitude of responses was seen in subject 161j, with combined CTL responses to all epitopes of >18,000 SFC/million PBMC. These responses are over 33-fold greater than those seen in subject 16732, who had the lowest total magnitude of responses. | FIG. 5Total magnitude of HIV-1-specific CTL responses for the eight HIV-1-infected subjects included in this study. The sum of the individual epitopes for each HLA class I A and B allele is indicated as follows: black bars, A2 epitopes; white bars, A3 epitopes; (more ...) |
The percent contribution of each HLA class I A and B allele to the total HIV-1-specific CTL response is indicated for each subject in Fig. 6. The relative contributions of each A and B allele were highly variable among all subjects. In subject AC-03 with acute HIV-1 infection, no responses were detected to any A2- or B7-restricted epitopes. In subjects 11324 and 13070, all of the HIV-1 epitopes recognized were restricted by either the A2, A3, or B7 allele. The response restricted by A2, which has been the most commonly studied HLA class I allele in terms of analyzing CTL responses in HIV-1 infection, was not the dominant contributor to the total CTL response with respect to the other three class I A and B alleles in any of the eight individuals studied. Subject 11841 exhibited the highest percentage of A2-restricted CTL responses, with 30% of total CTL responses detected in this study attributable to A2-restricted epitopes. However, in all seven of the other subjects, the contribution of the A2-restricted epitopes to the total CTL response ranged from 0% in subject AC-03 to 15% in subjects 221L and 161j. Hence, analysis of A2-restricted epitopes alone does not allow for sufficient representation of the total HIV-1-specific CTL response in an individual. These findings are consistent with previous studies among HLA-A2- and HIV-1-positive individuals by Betts et al. ( 5). | FIG. 6Relative contributions of each individual HLA A and B allele to the total HIV-1-specific CTL response in eight HIV-1-infected individuals, all expressing HLA A2, A3, and B7. Percentages were determined by dividing the total number of SFC/million PBMC (more ...) |
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REFERENCES 1. Addo, M M; Altfeld, M; Rosenberg, E S; Eldridge, R L; Philips, M N; Habeeb, K; Khatri, A; Brander, C; Robbins, G K; Mazzara, G P; Goulder, P J; Walker, B D. the HIV Controller Study Collaboration. The HIV-1 regulatory proteins Tat and Rev are frequently targeted by cytotoxic T lymphocytes derived from HIV-1-infected individuals. Proc Natl Acad Sci USA. 2001;98:1781–1786. [PubMed]2. Alexander-Miller, M A; Parker, K C; Tsukui, T; Pendleton, C D; Coligan, J E; Berzofsky, J A. Molecular analysis of presentation by HLA-A2.1 of a promiscuously binding V3 loop peptide from the HIV-envelope protein to human cytotoxic T lymphocytes. Int Immunol. 1996;8:641–649. [PubMed]3. Altfeld, M; Rosenberg, E S; Shankarappa, R; Mukherjee, J S; Hecht, F M; Eldridge, R L; Addo, M M; Poon, S H; Phillips, M N; Robbins, G K; Sax, P E; Boswell, S; Kahn, J O; Brander, C; Goulder, P J; Levy, J A; Mullins, J I; Walker, B D. Cellular immune responses and viral diversity in individuals treated during acute and early HIV-1 infection. J Exp Med. 2001;193:169–180. [PubMed]4. Bauer, M; Lucchiari-Hartz, M; Maier, R; Haas, G; Autran, B; Eichmann, K; Frank, R; Maier, B; Meyerhans, A. Structural constraints of HIV-1 Nef may curtail escape from HLA-B7-restricted CTL recognition. Immunol Lett. 1997;55:119–122. [PubMed]5. Betts, M R; Casazza, J P; Patterson, B A; Waldrop, S; Trigona, W; Fu, T M; Kern, F; Picker, L J; Koup, R A. Putative immunodominant human immunodeficiency virus-specific CD8 + T-cell responses cannot be predicted by major histocompatibility complex class I haplotype. J Virol. 2000;74:9144–9151. [PubMed]6. Borrow, P; Lewicki, H; Hahn, B H; Shaw, G M; Oldstone, M B. Virus-specific CD8 + cytotoxic T-lymphocyte activity associated with control of viremia in primary human immunodeficiency virus type 1 infection. J Virol. 1994;68:6103–6110. [PubMed]7. Borrow, P; Lewicki, H; Wei, X; Horwitz, M S; Peffer, N; Meyers, H; Nelson, J A; Gairin, J E; Hahn, B H; Oldstone, M B; Shaw, G M. Antiviral pressure exerted by HIV-1-specific cytotoxic T lymphocytes (CTLs) during primary infection demonstrated by rapid selection of CTL escape virus. Nat Med. 1997;3:205–211. [PubMed]8. Brander, C; Goulder, P J R. The evolving field of HIV CTL epitope mapping: new approaches to the identification of novel epitopes. In: Brander C, Korber B T M, Walker B D, Koup R A, Moore J, Haynes B, Meyers G. , editors. HIV molecular immunology database. Los Alamos, N.Mex: Los Alamos National Laboratory; 2000. 9. Brander, C; Walker, B D. The HLA class I restricted CTL response in HIV infection: systematic identification of optimal epitopes. In: Korber C B B, Walker B D, Koup R A, Moore J, Haynes B, Meyers G. , editors. HIV molecular immunology database. Los Alamos, N.Mex: Los Alamos National Laboratory; 1998. 10. Brander, C; Hartman, K E; Trocha, A K; Jones, N G; Johnson, R P; Korber, B; Wentworth, P; Buchbinder, S P; Wolinsky, S; Walker, B D; Kalams, S A. Lack of strong immune selection pressure by the immunodominant, HLA-A 0201-restricted cytotoxic T lymphocyte response in chronic human immunodeficiency virus-1 infection. J Clin Investig. 1998;101:2559–2566. [PubMed]11. Brander, C; Yang, O O; Jones, N G; Lee, Y; Goulder, P; Johnson, R P; Trocha, A; Colbert, D; Hay, C; Buchbinder, S; Bergmann, C C; Zweerink, H J; Wolinsky, S; Blattner, W A; Kalams, S A; Walker, B D. Efficient processing of the immunodominant, HLA-A 0201-restricted human immunodeficiency virus type 1 cytotoxic T-lymphocyte epitope despite multiple variations in the epitope flanking sequences. J Virol. 1999;73:10191–10198. [PubMed]12. Brander, C; Walker, B D. The HLA-class I restricted CTL response in HIV-1 infection: identification of optimal epitopes. In: Korber C B B, Walker B, Koup R, Haynes B, Moore J, Myers G. , editors. HIV-1 molecular immunology database. I. Theoretical biology and biophysics. Los Alamos, N.Mex: Los Alamos National Laboratory; 1995. 13. Bunce, M; Fanning, G C; Welsh, K I. Comprehensive, serologically equivalent DNA typing for HLA-B by PCR using sequence-specific primers (PCR-SSP). Tissue Antigens. 1995;45:81–90. [PubMed]14. Buseyne, F; McChesney, M; Porrot, F; Kovarik, S; Guy, B; Rivière, Y. Gag-specific cytotoxic T lymphocytes from human immunodeficiency virus type 1-infected individuals: Gag epitopes are clustered in three regions of the p24 gag protein. J Virol. 1993;67:694–702. [PubMed]15. Cao, H; Kanki, P; Sankale, J L; Dieng-Sarr, A; Mazzara, G P; Kalams, S A; Korber, B; Mboup, S; Walker, B D. Cytotoxic T-lymphocyte cross-reactivity among different human immunodeficiency virus type 1 clades: implications for vaccine development. J Virol. 1997;71:8615–8623. [PubMed]16. Culmann, B; Gomard, E; Kieny, M P; Guy, B; Dreyfus, F; Saimot, A G; Sereni, D; Sicard, D; Levy, J P. Six epitopes reacting with human cytotoxic CD8 + T cells in the central region of the HIV-1 NEF protein. J Immunol. 1991;146:1560–1565. [PubMed]17. Culmann-Penciolelli, B; Lamhamedi-Cherradi, S; Couillin, I; Guegan, N; Levy, J P; Guillet, J G; Gomard, E. Identification of multirestricted immunodominant regions recognized by cytolytic T lymphocytes in the human immunodeficiency virus type 1 Nef protein. J Virol. 1994;68:7336–7343. [PubMed]18. Del Val, M; Schlicht, H J; Ruppert, T; Reddehase, M J; Koszinowski, U H. Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein. Cell. 1991;66:1145–1153. [PubMed]19. Dupuis, M; Kundu, S K; Merigan, T C. Characterization of HLA-A 0201-restricted cytotoxic T cell epitopes in conserved regions of the HIV type 1 gp160 protein. J Immunol. 1995;155:2232–2239. [PubMed]20. Falk, K; Rotzschke, O; Stevanovic, S; Jung, G; Rammensee, H G. Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules. Nature. 1991;351:290–296. [PubMed]21. Goulder, P J; Addo, M M; Altfeld, M A; Rosenberg, E S; Tang, Y; Govender, U; Mngqundaniso, N; Annamalai, K; Vogel, T U; Hammond, M; Bunce, M; Coovadia, H M; Walker, B D. Rapid definition of five novel HLA-A 3002-restricted human immunodeficiency virus-specific cytotoxic T-lymphocyte epitopes by Elispot and intracellular cytokine staining assays. J Virol. 2001;75:1339–1347. [PubMed]22. Goulder, P J; Phillips, R E; Colbert, R A; McAdam, S; Ogg, G; Nowak, M A; Giangrande, P; Luzzi, G; Morgan, B; Edwards, A; McMichael, A J; Rowland-Jones, S. Late escape from an immunodominant cytotoxic T-lymphocyte response associated with progression to AIDS. Nat Med. 1997;3:212–217. [PubMed]23. Goulder, P J; Reid, S W; Price, D A; O'Callaghan, C A; McMichael, A J; Phillips, R E; Jones, E Y. Combined structural and immunological refinement of HIV-1 HLA-B8-restricted cytotoxic T lymphocyte epitopes. Eur J Immunol. 1997;27:1515–1521. [PubMed]24. Goulder, P J; Sewell, A K; Lalloo, D G; Price, D A; Whelan, J A; Evans, J; Taylor, G P; Luzzi, G; Giangrande, P; Phillips, R E; McMichael, A J. Patterns of immunodominance in HIV-1-specific cytotoxic T lymphocyte responses in two human histocompatibility leukocyte antigens (HLA)—identical siblings with HLA-A 0201 are influenced by epitope mutation. J Exp Med. 1997;185:1423–1433. [PubMed]25. Goulder, P J R; Brander, C; Annamalai, K; Mngqundaniso, N; Govender, U; Tang, Y; He, S; Hartman, K E; O'Callaghan, C A; Ogg, G S; Altfeld, M A; Rosenberg, E S; Cao, H; Kalams, S A; Hammond, M; Bunce, M; Pelton, S I; Burchett, S A; McIntosh, K; Coovadia, H M; Walker, B D. Differential narrow focusing of immunodominant human immunodeficiency virus Gag-specific cytotoxic T-lymphocyte responses in infected African and caucasoid adults and children. J Virol. 2000;74:5679–5690. [PubMed]26. Gray, C M; Lawrence, J; Schapiro, J M; Altman, J D; Winters, M A; Crompton, M; Loi, M; Kundu, S K; Davis, M M; Merigan, T C. Frequency of class I HLA-restricted anti-HIV CD8+ T cells in individuals receiving highly active antiretroviral therapy (HAART). J Immunol. 1999;162:1780–1788. [PubMed]27. Haas, G; Plikat, U; Debre, P; Lucchiari, M; Katlama, C; Dudoit, Y; Bonduelle, O; Bauer, M; Ihlenfeldt, H G; Jung, G; Maier, B; Meyerhans, A; Autran, B. Dynamics of viral variants in HIV-1 Nef and specific cytotoxic T lymphocytes in vivo. J Immunol. 1996;157:4212–4221. [PubMed]28. Haas, G; Samri, A; Gomard, E; Hosmalin, A; Duntze, J; Bouley, J M; Ihlenfeldt, H G; Katlama, C; Autran, B. Cytotoxic T-cell responses to HIV-1 reverse transcriptase, integrase and protease. AIDS. 1998;12:1427–1436. [PubMed]29. Harrer, E; Harrer, T; Barbosa, P; Feinberg, M; Johnson, R P; Buchbinder, S; Walker, B D. Recognition of the highly conserved YMDD region in the human immunodeficiency virus type 1 reverse transcriptase by HLA-A2-restricted cytotoxic T lymphocytes from an asymptomatic long-term nonprogressor. J Infect Dis. 1996;173:476–479. [PubMed]30. Jassoy, C; Johnson, R P; Navia, B A; Worth, J; Walker, B D. Detection of a vigorous HIV-1-specific cytotoxic T lymphocyte response in cerebrospinal fluid from infected persons with AIDS dementia complex. J Immunol. 1992;149:3113–3119. [PubMed]31. Jin, X; Bauer, D E; Tuttleton, S E; Lewin, S; Gettie, A; Blanchard, J; Irwin, C E; Safrit, J T; Mittler, J; Weinberger, L; Kostrikis, L G; Zhang, L; Perelson, A S; Ho, D D. Dramatic rise in plasma viremia after CD8 + T cell depletion in simian immunodeficiency virus-infected macaques. J Exp Med. 1999;189:991–998. [PubMed]32. Johnson, R P; Hammond, S A; Trocha, A; Siliciano, R F; Walker, B D. Induction of a major histocompatibility complex class I-restricted cytotoxic T-lymphocyte response to a highly conserved region of human immunodeficiency virus type 1 (HIV-1) gp120 in seronegative humans immunized with a candidate HIV-1 vaccine. J Virol. 1994;68:3145–3153. [PubMed]33. Johnson, R P; Trocha, A; Buchanan, T M; Walker, B D. Identification of overlapping HLA class I-restricted cytotoxic T cell epitopes in a conserved region of the human immunodeficiency virus type 1 envelope glycoprotein: definition of minimum epitopes and analysis of the effects of sequence variation. J Exp Med. 1992;175:961–971. [PubMed]34. Johnson, R P; Trocha, A; Yang, L; Mazzara, G P; Panicali, D L; Buchanan, T M; Walker, B D. HIV-1 gag-specific cytotoxic T lymphocytes recognize multiple highly conserved epitopes. Fine specificity of the gag-specific response defined by using unstimulated peripheral blood mononuclear cells and cloned effector cells. J Immunol. 1991;147:1512–1521. [PubMed]35. Kalams, S A; Goulder, P J; Shea, A K; Jones, N G; Trocha, A K; Ogg, G S; Walker, B D. Levels of human immunodeficiency virus type 1-specific cytotoxic T-lymphocyte effector and memory responses decline after suppression of viremia with highly active antiretroviral therapy. J Virol. 1999;73:6721–6728. [PubMed]36. Kalams, S A; Johnson, R P; Dynan, M J; Hartman, K E; Harrer, T; Harrer, E; Trocha, A K; Blattner, W A; Buchbinder, S P; Walker, B D. T cell receptor usage and fine specificity of human immunodeficiency virus 1-specific cytotoxic T lymphocyte clones: analysis of quasispecies recognition reveals a dominant response directed against a minor in vivo variant. J Exp Med. 1996;183:1669–1679. [PubMed]37. Koenig, S; Fuerst, T R; Wood, L V; Woods, R M; Suzich, J A; Jones, G M; de la Cruz, V F; Davey, R T, Jr; Venkatesan, S; Moss, B, et al. Mapping the fine specificity of a cytolytic T cell response to HIV-1 nef protein. J Immunol. 1990;145:127–135. [PubMed]38. Koup, R A; Safrit, J T; Cao, Y; Andrews, C A; McLeod, G; Borkowsky, W; Farthing, C; Ho, D D. Temporal association of cellular immune responses with the initial control of viremia in primary human immunodeficiency virus type 1 syndrome. J Virol. 1994;68:4650–4655. [PubMed]39. Lieberman, J; Fabry, J A; Kuo, M C; Earl, P; Moss, B; Skolnik, P R. Cytotoxic T lymphocytes from HIV-1 seropositive individuals recognize immunodominant epitopes in Gp160 and reverse transcriptase. J Immunol. 1992;148:2738–2747. [PubMed]40. McKinney, D M; Lewinsohn, D A; Riddell, S R; Greenberg, P D; Mosier, D E. The antiviral activity of HIV-specific CD8 + CTL clones is limited by elimination due to encounter with HIV-infected targets. J Immunol. 1999;163:861–867. [PubMed]41. Nixon, D F; Townsend, A R; Elvin, J G; Rizza, C R; Gallwey, J; McMichael, A J. HIV-1 gag-specific cytotoxic T lymphocytes defined with recombinant vaccinia virus and synthetic peptides. Nature. 1988;336:484–487. [PubMed]42. Ogg, G S; Jin, X; Bonhoeffer, S; Dunbar, P R; Nowak, M A; Monrad, S; Segal, J P; Cao, Y; Rowland-Jones, S L; Cerundolo, V; Hurley, A; Markowitz, M; Ho, D D; Nixon, D F; McMichael, A J. Quantitation of HIV-1-specific cytotoxic T lymphocytes and plasma load of viral RNA. Science. 1998;279:2103–2106. [PubMed]43. Pitcher, C J; Quittner, C; Peterson, D M; Connors, M; Koup, R A; Maino, V C; Picker, L J. HIV-1-specific CD4 + T cells are detectable in most individuals with active HIV-1 infection, but decline with prolonged viral suppression. Nat Med. 1999;5:518–525. [PubMed]44. Price, D A; Goulder, P J; Klenerman, P; Sewell, A K; Easterbrook, P J; Troop, M; Bangham, C R; Phillips, R E. Positive selection of HIV-1 cytotoxic T lymphocyte escape variants during primary infection. Proc Natl Acad Sci USA. 1997;94:1890–1895. [PubMed]45. Rowland-Jones, S L; Dong, T; Fowke, K R; Kimani, J; Krausa, P; Newell, H; Blanchard, T; Ariyoshi, K; Oyugi, J; Ngugi, E; Bwayo, J; MacDonald, K S; McMichael, A J; Plummer, F A. Cytotoxic T cell responses to multiple conserved HIV epitopes in HIV-resistant prostitutes in Nairobi. J Clin Investig. 1998;102:1758–1765. [PubMed]46. Rowland-Jones, S L; Powis, S H; Sutton, J; Mockridge, I; Gotch, F M; Murray, N; Hill, A B; Rosenberg, W M; Trowsdale, J; McMichael, A J. An antigen processing polymorphism revealed by HLA-B8-restricted cytotoxic T lymphocytes which does not correlate with TAP gene polymorphism. Eur J Immunol. 1993;23:1999–2004. [PubMed]47. Safrit, J T; Andrews, C A; Zhu, T; Ho, D D; Koup, R A. Characterization of human immunodeficiency virus type 1-specific cytotoxic T lymphocyte clones isolated during acute seroconversion: recognition of autologous virus sequences within a conserved immunodominant epitope. J Exp Med. 1994;179:463–472. [PubMed]48. Schmitz, J E; Kuroda, M J; Santra, S; Sasseville, V G; Simon, M A; Lifton, M A; Racz, P; Tenner-Racz, K; Dalesandro, M; Scallon, B J; Ghrayeb, J; Forman, M A; Montefiori, D C; Rieber, E P; Letvin, N L; Reimann, K A. Control of viremia in simian immunodeficiency virus infection by CD8 + lymphocytes. Science. 1999;283:857–860. [PubMed]49. Shankar, P; Fabry, J A; Fong, D M; Lieberman, J. Three regions of HIV-1 gp160 contain clusters of immunodominant CTL epitopes. Immunol Lett. 1996;52:23–30. [PubMed]50. Sidney, J; Grey, H M; Kubo, R T; Sette, A. Practical, biochemical and evolutionary implications of the discovery of HLA class I supermotifs. Immunol Today. 1996;17:261–266. [PubMed]51. Sipsas, N V; Kalams, S A; Trocha, A; He, S; Blattner, W A; Walker, B D; Johnson, R P. Identification of type-specific cytotoxic T lymphocyte responses to homologous viral proteins in laboratory workers accidentally infected with HIV-1. J Clin Investig. 1997;99:752–762. [PubMed]52. Suhrbier, A. Multi-epitope DNA vaccines. Immunol Cell Biol. 1997;75:402–408. [PubMed]53. Sutton, J; Rowland-Jones, S; Rosenberg, W; Nixon, D; Gotch, F; Gao, X M; Murray, N; Spoonas, A; Driscoll, P; Smith, M, et al. A sequence pattern for peptides presented to cytotoxic T lymphocytes by HLA B8 revealed by analysis of epitopes and eluted peptides. Eur J Immunol. 1993;23:447–453. [PubMed]54. Takahashi, K; Dai, L C; Fuerst, T R; Biddison, W E; Earl, P L; Moss, B; Ennis, F A. Specific lysis of human immunodeficiency virus type 1-infected cells by a HLA-A3.I-restricted CD8 + cytotoxic T-lymphocyte clone that recognizes a conserved peptide sequence within the gp41 subunit of the envelope protein. Proc Natl Acad Sci USA. 1991;88:10277–10281. [PubMed]55. Thomson, S A; Elliott, S L; Sherritt, M A; Sproat, K W; Coupar, B E; Scalzo, A A; Forbes, C A; Ladhams, A M; Mo, X Y; Tripp, R A; Doherty, P C; Moss, D J; Suhrbier, A. Recombinant polyepitope vaccines for the delivery of multiple CD8 cytotoxic T cell epitopes. J Immunol. 1996;157:822–826. [PubMed]56. Threlkeld, S C; Wentworth, P A; Kalams, S A; Wilkes, B M; Ruhl, D J; Keogh, E; Sidney, J; Southwood, S; Walker, B D; Sette, A. Degenerate and promiscuous recognition by CTL of peptides presented by the MHC class I A3-like superfamily: implications for vaccine development. J Immunol. 1997;159:1648–1657. [PubMed]57. Tsomides, T J; Aldovini, A; Johnson, R P; Walker, B D; Young, R A; Eisen, H N. Naturally processed viral peptides recognized by cytotoxic T lymphocytes on cells chronically infected by human immunodeficiency virus type 1. J Exp Med. 1994;180:1283–1293. [PubMed]58. Tsomides, T J; Walker, B D; Eisen, H N. An optimal viral peptide recognized by CD8 + T cells binds very tightly to the restricting class I major histocompatibility complex protein on intact cells but not to the purified class I protein. Proc Natl Acad Sci USA. 1991;88:11276–11280. [PubMed]59. Walker, B D; Flexner, C; Birch-Limberger, K; Fisher, L; Paradis, T J; Aldovini, A; Young, R; Moss, B; Schooley, R T. Long-term culture and fine specificity of human cytotoxic T-lymphocyte clones reactive with human immunodeficiency virus type 1. Proc Natl Acad Sci USA. 1989;86:9514–9518. [PubMed]60. Wilson, C C; Brown, R C; Korber, B T; Wilkes, B M; Ruhl, D J; Sakamoto, D; Kunstman, K; Luzuriaga, K; Hanson, I C; Widmayer, S M; Wiznia, A; Clapp, S; Ammann, A J; Koup, R A; Wolinsky, S M; Walker, B D. Frequent detection of escape from cytotoxic T-lymphocyte recognition in perinatal human immunodeficiency virus (HIV) type 1 transmission: the Ariel Project for the prevention of transmission of HIV from mother to infant. J Virol. 1999;73:3975–3985. [PubMed]61. Yang, O O; Walker, B D. CD8 + cells in human immunodeficiency virus type I pathogenesis: cytolytic and noncytolytic inhibition of viral replication. Adv Immunol. 1997;66:273–311. [PubMed]62. Yewdell, J W; Bennink, J R. Immunodominance in major histocompatibility complex class I-restricted T lymphocyte responses. Annu Rev Immunol. 1999;17:51–88. [PubMed] |