B细胞在抗CD45RB抗体诱导免疫耐受中的作用机制

发布时间：2018-05-16 08:13

本文选题：B淋巴细胞 + 免疫耐受　；参考：《暨南大学》2011年硕士论文

【摘要】：目的：本实验拟通过研究①体外CD45RB mAb处理过的B细胞对T细胞的作用；②体内B细胞缺失的情况下,抗CD45RB对CD3+CD45RBhiT细胞的影响；③抗CD45RB抗体处理的B细胞在体内诱导耐受；④抗CD45RB抗体诱导的免疫耐受中B细胞在体内的迁移状况等,阐述B淋巴细胞在抗CD45RB诱导的免疫耐受中可能发挥的作用机制。方法：在第0、1、3、5、7天分别向BALB/C裸鼠腹腔注射抗CD45RB抗体,7天后取脾制成单细胞悬液,与BALB/C小鼠T细胞、C57BL/6脾细胞混合培养,在第0、1、3、5、7天检测Thl, Th2, Treg, Tm。以B细胞缺陷鼠(B6μMT-/-)为受体建立皮肤移植模型,在移植后第0、1、3、5、7天,分别向受体鼠腹腔注射抗CD45RB单抗,在移植后第0、1、3、5、7、9天检测脾淋巴细胞CD3+CD45RBhi细胞比例。将BALB/C小鼠脾淋巴细胞、C57BL/6脾淋巴细胞混合培养,在第0、1、3、5、7天分别加入抗CD45RB抗体,培养7天后用尼龙毛柱分离淋巴细胞中的B细胞,给B细胞缺陷鼠(B6μMT-/-)通过尾静脉注入分离的B细胞,然后建立以B细胞缺陷鼠为受体的心脏移植模型,观察受体鼠生存期。通过CFSE将C57BL/6小鼠B细标记后,将标记的B细胞注入B细胞缺陷鼠体内,建立心脏移植模型,并在第0、1、3、5、7天分别向受体鼠腹腔注射抗CD45RB抗体,在第1、3、5、7、10天观察B细胞向胸腺的迁移情况。结果：在裸鼠体内用抗CD45RB抗体处理过的B细胞,与T细胞混合培养时,可显著Treg、Th2细胞比例明显升高,Thl细胞的比例下降,Tm细胞无明显影响。在体内B细胞缺失的情况下,抗CD45RB抗体依然能够降低T细胞表面CD45RB的表达,与对照组B细胞存在组相比,抗CD45RB抗体对T细胞表面CD45RB表达下调更为快速,但最终CD3+CD45RBhiT细胞比例无明显变化。体外抗CD45RB抗体处理过的B细胞可以延长受体的生存时间,但不能形成完全耐受。B6μMT-/-鼠在接受心脏移植和CFSE标记的B细胞后,第5天可以在胸腺内观察到标记的B细胞,并逐渐增多。对照组在第10天才在胸腺发现少量B细胞。结论：在抗CD45RB诱导的免疫耐受中,B细胞可能介导了对T细胞各亚群比例的影响,且在中枢耐受中也起到一定作用。但单独诱导B细胞的耐受只能延长移植物生存期,不能诱导完全耐受。B细胞缺失的情况下,抗CD45RB抗体对T细胞表面CD45RB表达下调更为快速,但与B细胞存在相比,最终CD3+CD45RBhiT细胞的比例是无明显差别的。
[Abstract]:Objective: to investigate the effect of CD45RB mAb on T cells in vitro and the effect of anti CD45RB on CD3 CD45RBhiT cells induced tolerance of B cells treated with anti CD45RB antibody in vivo. (4) the migration of B cells in vivo during the immune tolerance induced by anti CD45RB antibody. The possible mechanism of B lymphocytes in the immune tolerance induced by anti CD45RB was discussed. Methods: BALB/C nude mice were injected intraperitoneally with anti CD45RB antibody for 7 days. The spleen cells were mixed with T cells of BALB/C mice C57BL / 6. Thl, Th2, Tregand Tm were detected on the 7th day. The skin transplantation model was established with B cell deficient mice (B6 渭 MT-r -) as the receptor. On the 7th day after transplantation, anti CD45RB monoclonal antibodies were injected intraperitoneally into the recipient mice. The percentage of CD3 CD45RBhi cells in splenic lymphocytes was measured on day 0 1, 3 and 5 7 days after transplantation. Spleen lymphocytes of BALB/C mice were co-cultured with C57BL / 6 splenic lymphocytes. Anti CD45RB antibodies were added on the 7th day. After 7 days of culture, B cells in lymphocytes were separated by nylon capillary column. B cells were injected into B cells of B cell deficient mice by tail vein injection. Then the B-cell deficient rat heart transplantation model was established to observe the survival time of the recipient mice. After C57BL/6 mice B were labeled with CFSE, B cells were injected into B cell deficient mice to establish heart transplantation model. Anti CD45RB antibody was injected intraperitoneally into recipient mice on the 7th day of 0 ~ 1 ~ (th) ~ (th) day. The migration of B cells to the thymus was observed on the 7th day of the first trimester. Results: in nude mice treated with anti-THL antibody, the ratio of Treg-Th2 cells increased significantly when mixed with T cells. In the absence of B cells in vivo, anti CD45RB antibody can still reduce the expression of CD45RB on T cell surface. Compared with the control group, anti CD45RB antibody can down-regulate the expression of CD45RB on T cell surface more rapidly. But there was no significant change in the proportion of CD3 CD45RBhiT cells in the end. The B cells treated with anti CD45RB antibody in vitro could prolong the survival time of the receptor, but could not form completely tolerated. B6 渭 MT-r.-B cells could be observed in the thymus on the 5th day after heart transplantation and CFSE labeled B cells. And gradually increase. In the control group, a small number of B cells were found in the thymus on the 10th day. Conclusion: B cells may mediate the effect on T cell subsets in the immune tolerance induced by CD45RB and play a certain role in the central tolerance. However, the induction of B cell tolerance alone could only prolong the survival time of the grafts. However, when the completely tolerant. B cells were not induced, the down-regulation of CD45RB expression on T cell surface by anti CD45RB antibody was more rapid, but compared with the existence of B cell. Ultimately, there was no significant difference in the proportion of CD3 CD45RBhiT cells.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

【参考文献】