抗BLyS抗体抑制BLyS诱导小鼠脾细胞增殖实验方法的优化

Development of Anti-BLyS Antibody Neutralization Assay for BLyS-induced Splenocyte Proliferation

目的:建立BLyS抗体抑制BLyS作用下小鼠脾细胞增殖实验的方法。方法:首先通过DOE全因子实验验证BLyS能抑制脾淋巴细胞增殖后的凋亡,初步确定BLyS和刺激因子LPS的使用浓度、细胞密度及培养天数。然后进一步优化BLyS、Belimumab和LPS的使用浓度、孵育方式、培养天数等条件,确定抗体抑制BLyS作用下小鼠脾细胞增殖实验的方法。结果:BLyS能抑制小鼠脾淋巴细胞增殖后的凋亡;Belimumab对BLyS作用下小鼠脾细胞的增殖有显著中和作用。结论:建立BLyS抗体抑制BLyS作用下小鼠脾细胞增殖实验的方法,为进一步研究人BLyS与自身免疫疾病治疗药物奠定了基础。

Objectives： Establishing a method to detect the neutralization on BLyS-induced splenocyte proliferation. Methods： Design of experiment determine the initial concentration of BLyS, cell density and stimulation time. stimulation such as concentration of LPS, cell density and stimulation effect of anti-BLyS antibody （DOE） has been applied to Once the conditions of LPS time were determined, the concentrations of BLyS and Belimumab were optimized. Thereafter, the incubation time of Belimumab/BLyS were measured. Results： BLyS effectively inhibited the apoptosis of mouse splenocytes stimulated by LPS; Belimumab had a significant neutralization effect on BLyS-induced mouse splenocyte proliferation. Conclusion： It has been successfully established a method of measuring the inhibitory effect of anti-BLyS antibody against BLyS-induced splenocyte proliferation. It warrants future development of therapies for autoimmune diseases through inhibition of BLyS.