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Rapid identification of antigenic T‐cell epitopes by extracellular acidification rate signals
Author(s) -
Arimilli Subhashini,
Howard Maureen C.,
Nacy Carol C.,
Deshpande Shrikant V.
Publication year - 2000
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(20000601)77:3<409::aid-jcb6>3.0.co;2-#
Subject(s) - epitope , extracellular , t cell , t cell receptor , major histocompatibility complex , antigen , biology , cytotoxic t cell , interferon gamma , microbiology and biotechnology , immune system , chemistry , biochemistry , immunology , in vitro
We used a silicon‐based biosensor, a microphysiometer, to measure real‐time extracellular acidification rate signals associated with T lymphocyte responses to peptide ligands interacting with the T‐cell receptor (TCR). We compared these effector responses with those of interferon‐γ (IFN‐γ) production, and T‐cell proliferation. Within minutes, major histocompatibility complex (MHC)‐bound peptides on antigen‐presenting cells (APCs) engaged the TCR to increase acidification rates of the extracellular media was measured by microphysiometer. We exposed two myelin peptide‐specific human T‐cell clones, MSF132E11 (DRB1*1501 restricted) and TOM3A6 (DRB5*0101 restricted), to truncated analogues of the parent MBP 84‐102 peptide, in the presence of MHC restricted human antigen‐presenting cells, and measured the extracellular acidification rate signal changes, IFN‐γ production and T‐cell proliferation. The core epitopes recognized by these clones were identified by microphysiometer and found to be MBP 88–100 and MBP 91–100, respectively. These epitopes were identical to those identified by the IFN‐γ and proliferation assays. We conclude that measurement of real‐time extracellular acidification rate signals by the microphysiometer may facilitate rapid identification of human T‐cell epitopes involved in immune disorders and the development of specific T‐cell antagonists. J. Cell. Biochem. 77:409–417, 2000. © 2000 Wiley‐Liss, Inc.