| Literature DB >> 11150533 |
A Schmittel1, U Keilholz, S Bauer, U Kuhne, S Stevanovic, E Thiel, C Scheibenbogen.
Abstract
The ELISPOT assay has been established for the direct ex vivo quantification of peptide-reactive T lymphocytes from peripheral blood mononuclear cells (PBMC). In this report we studied the ELISPOT assay conditions for the detection of T cell responses against protein antigens including Tetanus toxoid (TT), purified-protein-derivative (PPD) and a synthetic 30 mer peptide derived from influenza matrix protein (IMP) containing the HLA-A*0201-restricted 9 mer peptide epitope GILGFVFTL as a model for a viral protein. We found several aspects to be crucial for a sensitive detection of T cell responses against proteins including a pellet preincubation step and the monocyte concentration. Using optimized assay conditions specific CD4+ T cell responses against TT and PPD as well as CD8+ T cell responses against IMP can be quantified directly ex vivo from peripheral blood mononuclear cells.Entities:
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Year: 2001 PMID： 11150533 DOI： 10.1016/s0022-1759(00)00305-7
Source DB: PubMed Journal: J Immunol Methods ISSN： 0022-1759 Impact factor: 2.303