T-cell responses to WT1 and proteinase 3 can be detected by ELISPOT assay.
Unstimulated PBMCs were analyzed for recognition of specific epitopes WT1 126-134 (A), WT1 187-194 (B), and proteinase 3 (C) by IFN-γ ELISPOT assay in 15 HLA-A2+ healthy subjects and in 15 AML patients. Values are expressed as number of spots per 106 PBMCs (white bars, number of spots in unstimulated PBMCs; black/striped bars, number of spots in response to peptide). Responses, which exceeded 2-fold or more the number of spots in unstimulated PBMCs and had a minimum of 10 peptide-specific spots in 106 PBMCs (after subtracting the number of spots in unstimulated PBMCs) were considered peptide specific and were shown as striped bars. The proportion of CD3 CD8 T cells in whole PBMCs, as determined by flow cytometry analyses, is indicated in AML patients (B).
