摘要
目的 对CD4 0配体CD15 4 (CD4 0L)在人B淋巴细胞中刺激转录因子NF κB活化进行研究。方法 应用重组CD15 4刺激EBV/LMP1阴性人RamosB细胞 ,观察对NF κBluciferase(萤光素酶 )活化的作用 ,判断CD15 4刺激对NF κB抑制蛋白IκB α、 β及 ε磷酸化和降解的影响 ,并对CD15 4刺激诱导进入细胞核NF κB亚单位进行分析。结果 CD15 4刺激 :(1)导致NF κBluciferase活性升高 ,且与CD15 4剂量正相关 ;(2 )引起细胞内IκB α、 β及 ε蛋白总量减少 ,IκB α、 β及 ε阳性细胞也明显减少 ;(3)主要诱导p5 0、p6 5及c Rel亚单位从细胞浆进入细胞核 ;(4)诱导p6 5磷酸化。结论 在人RamosB细胞中 ,CD15 4通过诱导IκB α、 β及 ε降解释放p5 0、p6 5及c Rel进入细胞核及p6 5磷酸化激活NF κB。
Objective To investigate the mechanism of CD15 4 -induced NF-κB activation in human B lymphocytes. Methods Using recombinant CD154 to stimulate EBV/LMP1 negative Ramos B cell to obser ve its effect in inducing NF-κB luciferase activity and degradation of IκB- α, -β, and -ε, and also to specify NF-κB subunits translocated into the n ucleus upon CD154 stimulation. Results CD154 stimulation of Ramos B cells induced degradation of IκB-α, -β, and -ε, phosphorylatio n of p65, binding of NF-κB subunits to DNA as shown by gel shift assay or by E LISA performed with nuclear extracts for the p50, p65 and c-Rel NF-κB subunit s and functional NF-κB activity as assessed by a luciferase reporter assay. Conclusion CD154-induced NF-κB activation in Ramos B cel ls requires degradation of IκB-α, -β, and -ε, releasing p50, p65 and c- Rel into the nucleus and also phosphorylation of p65. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第9期675-678,共4页
Chinese Journal of Microbiology and Immunology
