C. Scheibenbogen et al., Quantitation of antigen-reactive T cells in peripheral blood by IFN gamma-ELISPOT assay and chromium-release assay: a four-centre comparative trial, J IMMUNOL M, 244(1-2), 2000, pp. 81-89
The ELISPOT assay is increasingly being used for the monitoring of the indu
ction of antigen-reactive T cells in cancer vaccination trials. In order to
evaluate the reliability of T cell frequency analysis with the ELISPOT ass
ay, a comparative study was performed in four European laboratories. Six sa
mples from healthy subjects were analyzed for the frequency of influenza-re
active CD8+ T cells in peripheral blood mononuclear cells (PBMC) by IFN gam
ma -ELISPOT assay. In addition, one laboratory determined cytotoxic T cell
precursor (CTL) frequencies in these samples by limiting dilution chromium-
release assay (LDA), and three laboratories performed a variant of the LDA,
the multiple microculture assay (MMA). Consistent frequencies of influenza
peptide-reactive T cells were obtained with the ELISPOT assay in all four
laboratories. The numbers detected by ELISPOT assay correlated closely with
those determined by LDA. In contrast, the frequencies obtained with the MM
A differed considerably and showed Little correlation with the other two as
says. This study shows that it is possible to use the ELISPOT assay tl, det
ermine with reliability antigen-reactive T cells in a multicenter setting.
We suggest that this assay may be suitable for monitoring cancer vaccine tr
ials. (C) 2000 Elsevier Science B.V. All rights reserved.