Quantitation of antigen-reactive T cells in peripheral blood by IFNγ-ELISPOT assay and chromium-release assay: A four-centre comparative trial

Carmen Scheibenbogen, Pedro Romero, Licia Rivoltini, Wolfgang Herr, Alexander Schmittel, Jean Charles Cerottini, Thomas Woelfel, Alexander M M Eggermont, Ulrich Keilholz

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

The ELISPOT assay is increasingly being used for the monitoring of the induction of antigen-reactive T cells in cancer vaccination trials. In order to evaluate the reliability of T cell frequency analysis with the ELISPOT assay, a comparative study was performed in four European laboratories. Six samples from healthy subjects were analyzed for the frequency of influenza-reactive CD8+ T cells in peripheral blood mononuclear cells (PBMC) by IFNγ-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 MMA differed considerably and showed little correlation with the other two assays. This study shows that it is possible to use the ELISPOT assay to determine with reliability antigen-reactive T cells in a multicenter setting. We suggest that this assay may be suitable for monitoring cancer vaccine trials. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)81-89
Number of pages9
JournalJournal of Immunological Methods
Volume244
Issue number1-2
DOIs
Publication statusPublished - Oct 20 2000

Fingerprint

Enzyme-Linked Immunospot Assay
Chromium
T-Lymphocytes
Antigens
Human Influenza
T-Lymphoid Precursor Cells
Peptide T
Cancer Vaccines
Blood Cells
Healthy Volunteers
Vaccination
Neoplasms

Keywords

  • Chromium-release assay
  • ELISPOT assay
  • Limiting dilution assay

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Quantitation of antigen-reactive T cells in peripheral blood by IFNγ-ELISPOT assay and chromium-release assay : A four-centre comparative trial. / Scheibenbogen, Carmen; Romero, Pedro; Rivoltini, Licia; Herr, Wolfgang; Schmittel, Alexander; Cerottini, Jean Charles; Woelfel, Thomas; Eggermont, Alexander M M; Keilholz, Ulrich.

In: Journal of Immunological Methods, Vol. 244, No. 1-2, 20.10.2000, p. 81-89.

Research output: Contribution to journalArticle

Scheibenbogen, Carmen ; Romero, Pedro ; Rivoltini, Licia ; Herr, Wolfgang ; Schmittel, Alexander ; Cerottini, Jean Charles ; Woelfel, Thomas ; Eggermont, Alexander M M ; Keilholz, Ulrich. / Quantitation of antigen-reactive T cells in peripheral blood by IFNγ-ELISPOT assay and chromium-release assay : A four-centre comparative trial. In: Journal of Immunological Methods. 2000 ; Vol. 244, No. 1-2. pp. 81-89.
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