HLA-B57-restricted cytotoxic T-lymphocyte activity in a single infected subject toward two optimal epitopes, one of which is entirely contained within the other - PubMed (original) (raw)

HLA-B57-restricted cytotoxic T-lymphocyte activity in a single infected subject toward two optimal epitopes, one of which is entirely contained within the other

P J Goulder et al. J Virol. 2000 Jun.

Abstract

Viral peptides are recognized by cytotoxic T lymphocytes (CTL) as a complex with major histocompatibility complex (MHC) class I molecules, but the extent to which a single HLA allele can accommodate epitope peptides of different length and sequence is not well characterized. Here we report the identification of clonal CTL responses from the same donor that independently recognize one of two HLA-B57-restricted epitopes, KAFSPEVIPMF (KF11; p24(Gag) residues 30 to 40) and KAFSPEVI (KF8; p24(Gag) residues 30 to 37). Although lysis studies indicated that the KF11 peptide stabilized the HLA-B57-peptide complex more efficiently than the KI8 peptide, strong clonal responses were directed at each epitope. In samples from a second donor, the same phenomenon was observed, in which distinct CTL clones recognized peptide epitopes presented by the same HLA class I allele (in this case, HLA-A3) which were entirely overlapping. These data are relevant to the accurate characterization of CTL responses, which is fundamental to a detailed understanding of MHC class I-restricted immunity. In addition, these studies demonstrate marked differences in the length of peptides presented by HLA-B57, an allele which is associated with nonprogressive human immunodeficiency virus infection.

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Figures

FIG. 1

FIG. 1

(A) Patterns of recognition of peptides KI13 (KVIEEKAFSPEVI) and KF11 (KAFSPEVIPMF) by six representative CTL clones derived from donor 026-BMC. The peptide concentration was 100 μM, the effector-target cell (E:T) ratio in each case was 5:1, and the target cells were autologous EBV-transformed B cells (B-LCL). (B) Pattern of recognition of the same peptides by the same CTL clones as in panel A but using HLA-B57-matched targets from donor 003-BMC (HLA class I type of target cells, A23/—B45/57 Cw6/16; HLA class I type of 026-BMC CTL clones, A3/—B42/57 Cw7/17). The peptide concentration used was 10 μM, and the E:T ratio was 5:1. There was no recognition of target cells matched through class I molecules other than B57 or of HLA-mismatched targets (data not shown).

FIG. 2

FIG. 2

(A) Recognition of truncations of IK13 (KVIEEKAFSPEVI) and KF11 (KAFSPEVIPMF) by four representative CTL clones. The peptide concentration was 10 μM, the targets were B57-matched B-LCL from donor 003-BMC, and the E:T ratio was 5:1. (B) Recognition of truncations of KF11 at concentrations of peptide of 1, 10, and 100 μM by CTL clone 1 (similar pattern of recognition was observed for clone 2; not shown). Target cells were the same as in panel A. (C) Recognition of peptides differing by one amino acid at the N-terminal and C-terminal residues from the optimal peptide KI8 (KAFSPEVI) recognized by CTL clone 4 (a similar pattern of recognition was observed for clone 5; not shown) using concentrations of peptide of 1, 10, and 100 μM. Target cells were as in panel A. (D) Peptide titration curves showing specific recognition of KF11 by clone 1 and specific recognition of KI8 by clone 4. Target cells were as in panel A.

FIG. 2

FIG. 2

(A) Recognition of truncations of IK13 (KVIEEKAFSPEVI) and KF11 (KAFSPEVIPMF) by four representative CTL clones. The peptide concentration was 10 μM, the targets were B57-matched B-LCL from donor 003-BMC, and the E:T ratio was 5:1. (B) Recognition of truncations of KF11 at concentrations of peptide of 1, 10, and 100 μM by CTL clone 1 (similar pattern of recognition was observed for clone 2; not shown). Target cells were the same as in panel A. (C) Recognition of peptides differing by one amino acid at the N-terminal and C-terminal residues from the optimal peptide KI8 (KAFSPEVI) recognized by CTL clone 4 (a similar pattern of recognition was observed for clone 5; not shown) using concentrations of peptide of 1, 10, and 100 μM. Target cells were as in panel A. (D) Peptide titration curves showing specific recognition of KF11 by clone 1 and specific recognition of KI8 by clone 4. Target cells were as in panel A.

FIG. 3

FIG. 3

(A) Stabilization of peptide-B57 complexes by the peptides KF11 and KI8. Clone 1 (specific for KF11) shows recognition of KF11-pulsed targets >75 h after peptide pulsing. Clone 4 (specific for KI8) showed recognition of KI8-pulsed target cells for <18 h after peptide pulsing. The peptide concentration used was 10 μM. Targets and E:T ratios were as in Fig. 2. (B) Frequency of peptide-specific responses by Elispot assay following epitope optimization using fresh PBMC. The peptide concentration was 10 μM. Error bars show SD from the mean of four replicates for each peptide shown.

FIG. 4

FIG. 4

(A) Recognition of previously defined HLA-A3-restricted CTL epitopes RLRPGGKKK (RK9) and RLRPGGKKKY (RY10) by four clones derived from PBMC from donor 021-BMC (HLA A3/3001 B42/—Cw17/—). Targets were B-LCL from donor KS (HLA class I type A3/— B7/— Cw7/—) at an E:T ratio of 5:1. Specific lysis by the clones of targets pulsed with the peptide KIRLRPGGK (KK9) was between −1 and 0% (not shown). (B) Fine specificity of CTL clone 1. The best-recognized peptide was RY10. Similar data using CTL from clone 2 are not shown. Targets were the same as in panel A. (C) Fine specificity of CTL clone 3. The best-recognized peptide was RLRPGGKKKYK (RK11). Similar data using CTL from clone 4 are not shown. Targets were the same as in panel A.

FIG. 4

FIG. 4

(A) Recognition of previously defined HLA-A3-restricted CTL epitopes RLRPGGKKK (RK9) and RLRPGGKKKY (RY10) by four clones derived from PBMC from donor 021-BMC (HLA A3/3001 B42/—Cw17/—). Targets were B-LCL from donor KS (HLA class I type A3/— B7/— Cw7/—) at an E:T ratio of 5:1. Specific lysis by the clones of targets pulsed with the peptide KIRLRPGGK (KK9) was between −1 and 0% (not shown). (B) Fine specificity of CTL clone 1. The best-recognized peptide was RY10. Similar data using CTL from clone 2 are not shown. Targets were the same as in panel A. (C) Fine specificity of CTL clone 3. The best-recognized peptide was RLRPGGKKKYK (RK11). Similar data using CTL from clone 4 are not shown. Targets were the same as in panel A.

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