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. 1997 Sep 15;186(6):859–865. doi: 10.1084/jem.186.6.859

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(A) The majority of effectors detected in the ex vivo ELISPOT assay bear the memory-associated cell-surface marker CD45RO. Fresh PBMCs from donor SD were depleted of CD45RO⁺ cells and plated out at 5 × 10⁵/well with or without the HLA-B27.05–restricted peptide NP 383– 391. FACS^® analysis confirmed 67% depletion of CD45RO⁺ CD8⁺ T cells from PBMCs. (B) All detectable CD8⁺ effectors specific for the A2.01-restricted peptide M1 58–66 bear TCRs with the Vβ17 gene segment. The ELISPOT assay was performed with freshly isolated whole PBMCs or PBMCs depleted of Vβ17⁺ T cells; FACS^® analysis confirmed that the CD8⁺ Vβ17⁺ T cells initially present among whole PBMCs were depleted by 90%. Cells were seeded at 5 × 10⁵/well in the presence of the HLA-A2.01–restricted peptide M1 58–66 or the HLA-B8–restricted peptide NP 380–388. PBMCs were from donor WB (HLA class I haplotype: A1, A2.01; B7, B8).

(A) The majority of effectors detected in the ex vivo ELISPOT assay bear the memory-associated cell-surface marker CD45RO. Fresh PBMCs from donor SD were depleted of CD45RO⁺ cells and plated out at 5 × 10⁵/well with or without the HLA-B27.05–restricted peptide NP 383– 391. FACS^® analysis confirmed 67% depletion of CD45RO⁺ CD8⁺ T cells from PBMCs. (B) All detectable CD8⁺ effectors specific for the A2.01-restricted peptide M1 58–66 bear TCRs with the Vβ17 gene segment. The ELISPOT assay was performed with freshly isolated whole PBMCs or PBMCs depleted of Vβ17⁺ T cells; FACS^® analysis confirmed that the CD8⁺ Vβ17⁺ T cells initially present among whole PBMCs were depleted by 90%. Cells were seeded at 5 × 10⁵/well in the presence of the HLA-A2.01–restricted peptide M1 58–66 or the HLA-B8–restricted peptide NP 380–388. PBMCs were from donor WB (HLA class I haplotype: A1, A2.01; B7, B8).