摘要
目的：探讨CD86嵌合抗体ch1D1对系统性红斑狼疮（ systemic lupus erythematosus, SLE）患者自身反应性B细胞的作用及影响。方法利用流式细胞术分析SLE患者B细胞表面CD86的表达水平及ch1D1对SLE患者CD4＋T细胞活化的影响；利用磁珠法分选出健康志愿者和SLE患者外周血单个核细胞（ PBMC）中的B细胞、NK细胞以及CD4＋T细胞用于后续实验；利用LDH释放法检测ch1D1介导对SLE患者B细胞的抗体依赖的细胞介导的细胞毒作用（ antibody dependent cell medi-ated cytotoxicity,ADCC）及补体依赖的细胞介导的细胞毒作用（ complement dependent cell mediated cy-totoxicity,CDC）,利用ELISA法检测ch1D1对SLE患者B细胞分泌自身抗体的影响,利用3H掺入法分析ch1D1对SLE患者CD4＋T细胞增殖的影响。结果 SLE患者B细胞上CD80（68.08±14.28 vs 46.10±12.14,n=24,P〈0.0001）和CD86（44.72±14.90 vs 13.99±10.74,n=24,P〈0.0001）的表达水平显著高于健康志愿者,提示B细胞异常活化；与对照组相比,ch1D1能更有效介导对SLE患者B细胞的ADCC和CDC作用（P=0.0172,P=0.0388）；活化的T细胞显著增强SLE患者B细胞产生自身抗体,ch1D1显著抑制了SLE患者自身抗体的分泌（P=0.0019）；SLE患者CD4＋T细胞活化和增殖水平显著高于健康志愿者,而ch1D1显著抑制 SLE患者 CD4＋T细胞的增殖和活化（ P=0.0024,P=0.0495）。结论 CD86嵌合抗体能更有效地介导对SLE患者自身反应性B细胞的ADCC和CDC作用,抑制其自身抗体的分泌,抑制自身反应性CD4＋T细胞的增殖和活化,有望成为治疗SLE的新型免疫制剂。
Objective To investigate the effects of ch1D1, an anti-CD86 chimeric antibody, on autoreactive B lymphocytes isolated from patients with systemic lupus erythematosus （ SLE） . Methods Flow cytometry analysis was performed to measure the expression of CD86 on the surface of B cells isolated from patients with SLE and to analyze the effects of ch1D1 on the activation of CD4＋T cells. The method of magnetic bead sorting was used to separate B cells, NK cells and CD4＋T cells from PBMC collected from healthy subjects and patients with SLE for subsequent experiments. Antibody-dependent cell-mediated cyto-toxicity （ADCC） and complement-dependent cytotoxicity （CDC） that were mediated by ch1D1 were meas-ured with LDH release assay. Effects of ch1D1 on the secretion of auto-antibodies and the proliferation of CD 4＋ T were detected by ELISA and 3 H -thymidine （ 3 H-TdR） incorporation assay, respectively. Results The levels of CD80 （68. 08±14. 28 vs 46. 10±12. 14, n=24, P〈0. 000 1） and CD86 （44. 72±14. 90 vsnbsp;13. 99±10. 74, n=24, P〈0. 000 1） expressed on the surface of B cells isolated from patients with SLE were significantly higher than those from the healthy subjects, suggesting the abnormal activation of B cells. Com-pared with the negative control group and the murine monoclonal antibody 1D1, ch1D1 was more effective in mediating the ADCC and CDC responses （P=0. 017 2, P=0. 038 8）. Activated T cells significantly en-hanced the secretion of auto-antibodies by B cells isolated from patients with SLE. Compared with the nega-tive control group, the enhanced secretion of auto-antibodies was significantly inhibited by treatment with ch1D1 （P=0. 001 9）. Moreover, ch1D1 significantly inhibited the proliferation and activation of CD4＋T cells induced in patients with SLE （P=0. 002 4, P=0. 049 5）. Conclusion ch1D1, the anti-CD86 chim-eric antibody, could effectively mediate the ADCC and CDC responses against autoreactive B cells isolated from patients with SLE, inhibit the secretion of auto-antibodies and suppress the proliferation and activation of auto-reactive CD4＋T cells. It might be a potential immunotherapy agent for the treatment of SLE.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2016年第7期481-486,共6页
Chinese Journal of Microbiology and Immunology
基金
杭州市科技发展计划项目（20120533Q29）
浙江省自然科学基金（LY15C080001）
杭州市医药卫生科技计划项目（20108010）
作者简介
通信作者：刘杨，Email：liuyang0620@zj．edu．cn，电话：0571—88208286
