DETECTION OF INTRACELLULAR INTERFERON-GAMMA BY LIGHT-MICROSCOPY USING AN IMMUNOPEROXIDASE TECHNIQUE - CORRELATION WITH THE CORRESPONDING MESSENGER-RNA AND PROTEIN PRODUCT

被引:30
作者
DOLHAIN, RJEM
ANDERSSON, U
TERHAAR, NT
BRINKMAN, BMN
VERWEIJ, CL
DAHA, MR
BREEDVELD, FC
MILTENBURG, AMM
机构
[1] UNIV HOSP LEIDEN,DEPT NEPHROL,2300 RC LEIDEN,NETHERLANDS
[2] UNIV STOCKHOLM,ARRHENIUS LABS NAT SCI,DEPT IMMUNOL,S-10691 STOCKHOLM,SWEDEN
关键词
IMMUNOCYTOCHEMISTRY; IMMUNOFLUORESCENCE; POLYMERASE CHAIN REACTION; ELISA;
D O I
10.1002/jlb.54.6.545
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Identifying individual cytokine-producing cells may help to acquire insight into immunological processes. This study was designed to adapt a technique for the detection of individual cytokine-producing cells from an immunofluorescence to an immunoperoxidase staining procedure. The production of interferon-gamma (IFN-gamma) by anti-CD3-activated cloned human T cells was used as a model system. After the conditions for the staining procedure were optimized, the immunoperoxidase technique was slightly more sensitive than the immunofluorescence technique. The intracellular staining for IFN-gamma was preceded or paralleled by IFN-gamma mRNA production and followed by accumulation of IFN-gamma in the supernatant. It is concluded that intracellular IFN-gamma can easily be detected using an immunoperoxidase procedure. This procedure is highly sensitive and allows quantification of the production of multiple cytokines by counting the percentage of positively staining cells.
引用
收藏
页码:545 / 551
页数:7
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