6533b821fe1ef96bd127ba94
RESEARCH PRODUCT
Quantitation of antigen-reactive T cells in peripheral blood by IFNgamma-ELISPOT assay and chromium-release assay: a four-centre comparative trial
Licia RivoltiniUlrich KeilholzWolfgang HerrCarmen ScheibenbogenAlexander M.m. EggermontPedro RomeroAlexander SchmittelThomas WoelfelJean Charles Cerottinisubject
T cellImmunologyEpitopes T-LymphocyteIndicator Dilution TechniquesEnzyme-Linked Immunosorbent AssayCD8-Positive T-LymphocytesLymphocyte ActivationPeripheral blood mononuclear cellViral Matrix ProteinsInterferon-gammaAntigenHLA-A2 AntigenHumansImmunology and AllergyCytotoxic T cellMedicineAntigens ViralImmunodominant Epitopesbusiness.industryELISPOTMolecular biologyChromium RadioisotopesHIV Reverse TranscriptasePeptide FragmentsCTL*medicine.anatomical_structureImmunologyLeukocytes MononuclearCancer vaccinebusinessCD8T-Lymphocytes Cytotoxicdescription
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 IFNgamma-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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2000-01-01 |