Monitoring of CD95 (APO-1/Fas) ligand expression in human T cells by quantitative RT-PCR.
CD95 (APO-1/Fas) receptor/ligand interaction is a key regulatory pathway for apoptosis in lymphoid cells. We developed a quantitative RT-PCR for the human CD95-L to determine expression levels in lymphoid cell lines and in lymphocytes derived from blood of healthy individuals. In untreated peripheral blood T lymphocytes and T cell lines constitutive expression of the CD95-L mRNA was found at low levels. Stimulation of T cells by treatment with PMA and ionomycine (P/I) lead up to a 100-fold maximal increase in CD95-L mRNA after 4 h. CD95-L mRNA is produced by activated CD8 and CD4T cells. In vivo increased CD95-L mRNA expression was found in freshly isolated T cells during the acute phase of EBV infection. In contrast to T cells, CD95-L mRNA could be induced in some B lineage cell lines only after five days of stimulation. Since defective or accelerated CD95/CD95-L interaction is considered to be involved in the pathogenesis of lymphoproliferation, autoimmunity and AIDS, the quantitative RT-PCR assay described in this paper may provide a powerful tool for monitoring CD95-L expression in these diseases.
Herr
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Hermatology/Oncology, University Children's Hospital and Division of Molecular Oncology, German Cancer Research Center, Heidelberg, Germany.
Balemans
L
L
Böhler
T
T
Walczak
H
H
Debatin
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KM