Table I.

Frequency, surface phenotype, and IFN-γ secretion by Ag-specific CD8+ T lymphocytes

IndividualFrequencyaCD45RAbCCR7bSecretion Assayc
HIV-1 AgCognate Ag
Melan-A
HD 00090.18100ND10
HD 27060.04909500
HD 27100.05959000
HD 27110.0590ND00
HD 27130.0485ND00
HD 75900.09959511
HD 78340.1010010011
Mean0.08959500
Influenza
HD 27140.13205508
HD 27150.23105062
HD 42960.171050014
HD 44690.15030049
HD 44700.072540ND27
HD 75900.093585ND15
HD 78330.48901010
Mean0.192540025
CMV
HD 27091.9650066
HD 27134.5510279
HD 44690.9655ND25
HD 44701.55010045
HD 44741.4155032
HD 75172.9355158
HD 752412.2555172
HD 78331.455377
HD 78471.17015046
Mean3.2405156
  • a CD8+ lymphocyte populations were highly purified from PBMC (>98%) and stained with the appropriate PE-labeled tetramers along with anti-CD8PerCP mAb. Values are the percentage of A2/tetramer+ cells in gated CD8+ cells.

  • b CD8+ cells were also stained with anti-CD45RAAPC mAb or rat anti-CCR7 mAb labeled with goat anti-rat IgGFITC. Values are the percentage of A2/tetramer+ cells with the corresponding phenotype.

  • c PBMC were stimulated for 4 h with 1 μg/ml irrelevant HIV-1 peptide or cognate peptide and stained with the appropriate APC-labeled A2/tetramers. Cell surface detection of IFN-γ-secreted molecules was performed by using an IFN-γ-specific affinity matrix and subsequent labeling with PE-conjugated anti-IFN-γ mAb. Values are the percentage of IFN-γ+ cells in gated A2/tetramer+ CD8+ cells.