Molecular and functional analysis of a conserved CTL epitope in HIV-1 p24 recognized from a long-term nonprogressor: constraints on immune escape associated with targeting a sequence essential for viral replication.
It has been hypothesized that sequence variation within CTL epitopes leading to immune escape plays a role in the progression of HIV-1 infection. Only very limited data exist that address the influence of biologic characteristics of CTL epitopes on the emergence of immune escape variants and the efficiency of suppression HIV-1 by CTL. In this report, we studied the effects of HIV-1 CTL epitope sequence variation on HIV-1 replication. The highly conserved HLA-B14-restricted CTL epitope DRFYKTLRAE in HIV-1 p24 was examined, which had been defined as the immunodominant CTL epitope in a long-term nonprogressing individual. We generated a set of viral mutants on an HX10 background differing by a single conservative or nonconservative amino acid substitution at each of the P1 to P9 amino acid residues of the epitope. All of the nonconservative amino acid substitutions abolished viral infectivity and only 5 of 10 conservative changes yielded replication-competent virus. Recognition of these epitope sequence variants by CTL was tested using synthetic peptides. All mutations that abrogated CTL recognition strongly impaired viral replication, and all replication-competent viral variants were recognized by CTL, although some variants with a lower efficiency. Our data indicate that this CTL epitope is located within a viral sequence essential for viral replication. Targeting CTL epitopes within functionally important regions of the HIV-1 genome could limit the chance of immune evasion.
Wagner
R
R
Institute of Medical Microbiology, University of Regensburg, Germany.
Leschonsky
B
B
Harrer
E
E
Paulus
C
C
Weber
C
C
Walker
B D
BD
Buchbinder
S
S
Wolf
H
H
Kalden
J R
JR
Harrer
T
T
D000818:Animals; D000940:Antigenic Variation / Q000235:genetics; D019556:COS Cells; D002461:Cell Line, Transformed; D017124:Conserved Sequence / Q000276:immunology*; D018450:Disease Progression; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D016655:HIV Core Protein p24 / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007108:Immune Tolerance / Q000235:genetics; D008297:Male; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D011533:Proviruses / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication / Q000235:genetics / Q000276:immunology*
D018984:Epitopes, T-Lymphocyte; D016655:HIV Core Protein p24; D010446:Peptide Fragments