Stimulation of TNF-β and IL-2 production by PMA/PHA requires p21ras, raf and ERK
- VernacularTitle:癌基因p21ras、raf和细胞外信号调节蛋白激酶参与刺激Jurkat细胞产生TNF-β和IL-2
- Author:
Tiesheng HUANG
;
Yunzhu HUANG
- Publication Type:Journal Article
- From:
Chinese Journal of Microbiology and Immunology
2001;21(2):134-137
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether the p21ras/ERK (MAPK) cascade is involved in regulating the production of TNF-β and IL-2 in Jurkat cells. Methods Cells were transiently transfected with a dominant negation construct of p21ras (ras15A or ras17N), raf (raf 1-130) or ERK1 (erk1K71R), or transforming p21ras (H-ras61L) or constitutively active raf (raf22W) using lipofectamine. Control cells received a plasmid carrying β-galactosidease. Transfectant cells were stimulated with PMA/PHA and released TNF-β and IL-2 quantitated by ELISA. Results In control cells, the production of TNF-β and IL-2 was approx. 2 fold by PMA/PHA stimulation (P<0.001), while transfection with a dominant negative mutant construct abrogated the effects of PMA/PHA. Transfection with a constitutively active mutant had no effect on basal production of TNF-β or IL-2, but all of the constitutively active mutants significantly enhanced the response of the cells to PMA/PHA (P<0.05). MEK1 inhibitor PD98059 inhibited the effects of PMA/PHA and active construct genes on ERK activities and cytokine production. Conclusion These data demonstrate that the p21ras-ERK cascade plays an important role in regulating the production of both TNF-β and IL-2 in Jurkat cells stimulated with PMA/PHA.
