[The enhancement effect of CD59 on CD55-mediated T cell signal transduction].
Fei Liao, Meihua Gao, Bei Zhan
Abstract
Fei Liao, Meihua Gao, Bei Zhan
Abstract
AIM: To study the enhancement effect of glycosylphosphatidyl inositol (GPI)-anchored protein CD59 on CD55-mediated T cell signal transduction. METHODS: Human Jurkat cells were divided into 3 groups: Jurkat cells(Igroup), Jurkat cells transfected with blank plasmid(II group)and Jurkat cells transfected with siRNA plasmid (III group). CD59 mRNA level was detected by RT-PCR. The cell proliferation activity of the three groups was measured by MTTcolorimetry after crosslinking anti-CD55 mAb and anti-CD59 mAb. The phosphorylation levels of Src family of protein tyrosine kinase (PTK) were investigated by immunoblot analysis, and the kinetic changes of [Ca(2+);] in T cell endochylema were determined by Laser scanning confocal microscope imaging. RESULTS: CD59 expression was successfullyinhibited in IIIgroup cells after transfecting. The cell proliferation, phosphorylation levels of Src family of PTK and degree of [Ca(2+);] inIand IIgroups were increased compared with III group(P<0.05), after crosslinking of CD55 and CD59 antibodies, but there was no difference between I group and II group. CONCLUSION: CD59 can enhance the effect of CD55-mediated T cell signal transduction.
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AIM: To study the enhancement effect of glycosylphosphatidyl inositol (GPI)-anchored protein CD59 on CD55-mediated T cell signal transduction. METHODS: Human Jurkat cells were divided into 3 groups: Jurkat cells(Igroup), Jurkat cells transfected with blank plasmid(II group)and Jurkat cells transfected with siRNA plasmid (III group). CD59 mRNA level was detected by RT-PCR. The cell proliferation activity of the three groups was measured by MTTcolorimetry after crosslinking anti-CD55 mAb and anti-CD59 mAb. The phosphorylation levels of Src family of protein tyrosine kinase (PTK) were investigated by immunoblot analysis, and the kinetic changes of [Ca(2+);] in T cell endochylema were determined by Laser scanning confocal microscope imaging. RESULTS: CD59 expression was successfullyinhibited in IIIgroup cells after transfecting. The cell proliferation, phosphorylation levels of Src family of PTK and degree of [Ca(2+);] inIand IIgroups were increased compared with III group(P<0.05), after crosslinking of CD55 and CD59 antibodies, but there was no difference between I group and II group. CONCLUSION: CD59 can enhance the effect of CD55-mediated T cell signal transduction.
Key concepts: Jurkat cells, CD59, Molecular biology, Transfection, Signal transduction, Cell growth, Tyrosine phosphorylation, Phosphorylation