摘要
目的探讨钙调神经磷酸酶(Calcineurin,CaN)对金黄色葡萄球菌肠毒素C2(staphylococcal enterotoxin C2,SEC2)发挥超抗原活性的作用。方法以终浓度为0.5、1、2、3和4μg/ml的重组野生型rSEC2及其增强型突变体蛋白mSEC2(T20L/G22E/H118A/H122A)刺激SD大鼠脾淋巴细胞,MTT法检测大鼠脾淋巴细胞增殖水平;检测终浓度为1μg/ml的rSEC2和mSEC2在诱导大鼠脾淋巴细胞增殖过程中CaN活性的变化、终浓度为0.1、0.5、1、2和3μg/ml的CaN特异性抑制剂环孢菌素A(cyclosporine A,CsA)对该过程的影响,以及CaN3个亚基CaN Aα、CaN Aβ、CaN B在mRNA水平上的变化。结果在低浓度rSEC2和mSEC2刺激下,大鼠脾淋巴细胞增殖指数(proliferation index,PI)与给药浓度成正比,高浓度刺激下,可明显抑制大鼠脾淋巴细胞的增殖,1μg/ml浓度增殖效果最好;CsA对大鼠脾淋巴细胞增殖具有较强的抑制作用,呈明显的剂量依赖性,当CsA浓度为0.5～1μg/ml时,rSEC2和mSEC2的刺激作用即被完全抑制;CaN的活性及其相关亚基(CaN Aβ和CaN B)mRNA水平与其超抗原的刺激增殖活性具有明显相关性。结论 CaN对SEC2发挥超抗原活性具有重要作用,为提高SEC2的临床应用价值及规避毒副作用奠定了理论基础。
Objective To investigate the effect of calcineurin （CAN） on the superantigne activity of staphylococcal en- terotoxin C2 （SEC2）. Methods Splenic lymphocytes of SD rats were stimulated with recombinant wild SEC2 （rSEC2） and its enhanced mutant （mSEC2） （T20L/G22E / H118A/H122A）, at various final concentrations （0. 5, 1, 2, 3 and 4 μg / ml）, and determined for proliferation level by MTF assay. The changes of CaN activity in the course of induction of proliferation of rat lymphocytes with rSEC2 and mSEC2, at a final concentration of 1 μg / ml, were determined, while the effect of various final concentrations （0. 1, 0. 5. 1, 2 and 3 μg/ml） of cyclosporine A （CsA）, a specific inhibitor of CaN, on the course, as well as the changes of three CaN subunits （CaN Act, CaN AI3 and CaN B） at mRNA level, were evaluated. Results The proliferation index （PI） of rat splenic lymphocytes stimulated with low concentration rSEC2 and mSEC2 was dose-dependent. However, high rSEC2 and mSEC2 concentrations inhibited the proliferation significantly, of which the those at a concentration of 1 μg/ml showed a satisfactory inhibitory effect. CsA showed strong dose-dependent inhibitory effect on the proliferation of rat splenic lymphocytes, of which those at a concentration of 0. 5 - 1 μg / ml com- pletely inhibited the stimulating effect of rSEC2 and mSEC2. CaN activity as well as CaN Aβ and CaN B mRNA levels were significantly related to the superantigen activity of CaN. Conclusion CaN played an important role in superantigen activity of SEC2, which laid a theoretical basis for enhancing the clinical application and avoiding the adverse reaction of SEC2.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第10期1413-1417,共5页
Chinese Journal of Biologicals
基金
国家科技重大专项重大新药创制(2012ZX09102301-013)
沈阳市科技计划项目(F11-264-1-11
F12-152-9-00)资助
