RRC ID 39262
著者 Okubo M, Brown MP, Chiba K, Kasukawa R, Nishimaki T.
タイトル Detection of TNF alpha and Fas ligand mRNA within synovial mononuclear cells by fluorescence in-cell labeling PCR (FICL-PCR).
ジャーナル Mol Biol Rep
Abstract T cells that infiltrate the synovial lesions of rheumatoid arthritis may play a key role in its pathogenesis. To learn more about their functional nature, we determined the frequency of synovial T cells that harbored the TNF alpha and Fas ligand transcript by a technique, called Fluorescence In-Cell Labeling Polymerase Chain Reaction (FICL-PCR). The mRNA of interest was detected in fixed cells by the incorporation during PCR of a fluorescein-12-dUTP label following an initial reverse transcription PCR step. Using this technique the CD3 transcript was detected in the T leukemic cell line, MOLT-4, with calculated sensitivity and specificity values of 91% and 100%, respectively. The percentage mean (+/-S.D.) of TNF alpha mRNA positive cells and Fas ligand mRNA positive cells in peripheral blood mononuclear cells from 12 rheumatoid arthritis patients were 5.1+/-2.3% and 4.8+/-3.1%, respectively. The percentage mean (+/-S.D.) of TNF alpha mRNA positive cells and Fas ligand mRNA positive cells among synovial mononuclear cells from six rheumatoid arthritis patients was 16.8+/-8.3% and 10.8+/-1.8%, respectively. This result indicates that the cytotoxic T cells expressing TNF alpha accumulate in rheumatoid arthritic lesions where they may play a pathogenic role.
巻・号 25(4)
ページ 217-24
公開日 1998-11-1
DOI 10.1023/a:1006863303503
PMID 9870611
MeSH Arthritis, Rheumatoid / immunology CD3 Complex / metabolism Fas Ligand Protein Fluorescence Humans Ligands Membrane Glycoproteins / genetics Membrane Glycoproteins / metabolism* Molecular Sequence Data RNA, Messenger / genetics RNA, Messenger / metabolism Reverse Transcriptase Polymerase Chain Reaction / methods* Sensitivity and Specificity Synovial Fluid / cytology* Synovial Fluid / immunology T-Lymphocytes / immunology T-Lymphocytes / metabolism* Tumor Necrosis Factor-alpha / genetics Tumor Necrosis Factor-alpha / metabolism*
IF 1.402
引用数 8
リソース情報
ヒト・動物細胞 MOLT-4(RCB0206) DAUDI(RCB1640)