支架蛋白JLP调控B细胞上CD40诱导的MAPK信号通路活化及细胞增殖

目的 对支架蛋白JLP在小鼠B细胞上CD40诱导信号通路活化及细胞增殖中的作用进行研究,探索支架蛋白JLP在CD40诱导小鼠B细胞信号通路及后续生物学效应中的作用.方法 磁珠分选法分选出jlp野生型和jlp基因敲除小鼠模型脾脏B细胞,使用免疫印迹法检测在接受CD40配体（CD40L）刺激后jlp野生型和jlp基因敲除型小鼠脾脏B细胞p38、Erk和Jnk MAPK,磷酸化c-Jun以及NF-κB信号通路及的活化情况,并使用CFSE检测两种基因型小鼠B细胞在接受CD40L刺激后细胞增殖情况.结果 jlp基因敲除小鼠B细胞在受到CD40L刺激后其p38、Erk、Jnk MAPK信号通路以及c-Jun活化情况均较jlp野生型小鼠B细胞明显减弱,而NF-κB信号通路的激活情况在两种基因型B细胞中并无明显差异,jlp基因敲除小鼠B细胞中CD40诱导的细胞增殖水平较jlp野生型B细胞明显降低.结论 支架蛋白JLP参与调控B细胞上CD40诱导的MAPK信号通路活化及细胞增殖.

Scaffolding Protein JLP Regulates CD40L-Induced MAPK Signal Pathway Activity and Cell Proliferation in B Lymphocyte

Objective To investigate the role of scaffolding protein JLP in regulating CD40L - induced activation of signal pathways and B cell proliferation, thus to explore whether JLP is involved in CD40L - triggered signals activity and downstream biological outcomes. Methods B lymphocyte were purified from splenocytes from jlp ＋/＋ and jlp -/- mice by magnetic sorting for the negative selection. Western blotting was used for detecting the p38, Erk, Jnk MAPK, c - Jun and IK -Bct phosphorylation. CFSE was performed to analyze the proliferation of two genotypic B cells. Results The phosphorylated forms of p38, Erk and Jnk were impaired in jlp -/- splenic B cells comparing with control B cells upon CD40 treatment. CD40 engagement also resulted in a marg - inal decrease in c - Jun phosphorylation in jlp-/- B cells. However, no substantial differences in the level of phosphorylated IkBa could be observed between two genotypic of B cells. Conclusion Scaffolding protein JLP is involved in regulating CD40 - triggered activation of MAPK signal pathway and the proliferation in B lymphocyte.