Application of the IFN-γ ELISPOT assay to quantify T cell responses against proteins

被引：58
作者：
Schmittel, A
Keilholz, U
Bauer, S
Kuhne, U
Stevanovic, S
Thiel, E
Scheibenbogen, C
机构：
[1] Free Univ Berlin, Klinikum Benjamin Franklin, Med Klin 3, D-12200 Berlin, Germany
[2] Univ Tubingen, Inst Zelibiol, Immunol Abt, D-72076 Tubingen, Germany
来源：
JOURNAL OF IMMUNOLOGICAL METHODS | 2001年 / 247卷 / 1-2期
关键词：
ELISPOT; T cells; protein; tetanus toxoid; purified protein derivative; influenza matrix protein;
D O I：
10.1016/S0022-1759(00)00305-7
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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 toroid (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. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
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页码：17 / 24
页数：8
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