發(fā)布時間：2018-05-16 08:13

  本文選題：B淋巴細胞 + 免疫耐受��； 參考：《暨南大學》2011年碩士論文

【摘要】：目的：本實驗擬通過研究①體外CD45RB mAb處理過的B細胞對T細胞的作用；②體內(nèi)B細胞缺失的情況下,抗CD45RB對CD3+CD45RBhiT細胞的影響；③抗CD45RB抗體處理的B細胞在體內(nèi)誘導耐受；④抗CD45RB抗體誘導的免疫耐受中B細胞在體內(nèi)的遷移狀況等,闡述B淋巴細胞在抗CD45RB誘導的免疫耐受中可能發(fā)揮的作用機制。 方法：在第0、1、3、5、7天分別向BALB/C裸鼠腹腔注射抗CD45RB抗體,7天后取脾制成單細胞懸液,與BALB/C小鼠T細胞、C57BL/6脾細胞混合培養(yǎng),在第0、1、3、5、7天檢測Thl, Th2, Treg, Tm。以B細胞缺陷鼠(B6μMT-/-)為受體建立皮膚移植模型,在移植后第0、1、3、5、7天,分別向受體鼠腹腔注射抗CD45RB單抗,在移植后第0、1、3、5、7、9天檢測脾淋巴細胞CD3+CD45RBhi細胞比例。將BALB/C小鼠脾淋巴細胞、C57BL/6脾淋巴細胞混合培養(yǎng),在第0、1、3、5、7天分別加入抗CD45RB抗體,培養(yǎng)7天后用尼龍毛柱分離淋巴細胞中的B細胞,給B細胞缺陷鼠(B6μMT-/-)通過尾靜脈注入分離的B細胞,然后建立以B細胞缺陷鼠為受體的心臟移植模型,觀察受體鼠生存期。通過CFSE將C57BL/6小鼠B細標記后,將標記的B細胞注入B細胞缺陷鼠體內(nèi),建立心臟移植模型,并在第0、1、3、5、7天分別向受體鼠腹腔注射抗CD45RB抗體,在第1、3、5、7、10天觀察B細胞向胸腺的遷移情況。 結(jié)果：在裸鼠體內(nèi)用抗CD45RB抗體處理過的B細胞,與T細胞混合培養(yǎng)時,可顯著Treg、Th2細胞比例明顯升高,Thl細胞的比例下降,Tm細胞無明顯影響。在體內(nèi)B細胞缺失的情況下,抗CD45RB抗體依然能夠降低T細胞表面CD45RB的表達,與對照組B細胞存在組相比,抗CD45RB抗體對T細胞表面CD45RB表達下調(diào)更為快速,但最終CD3+CD45RBhiT細胞比例無明顯變化。體外抗CD45RB抗體處理過的B細胞可以延長受體的生存時間,但不能形成完全耐受。B6μMT-/-鼠在接受心臟移植和CFSE標記的B細胞后,第5天可以在胸腺內(nèi)觀察到標記的B細胞,并逐漸增多。對照組在第10天才在胸腺發(fā)現(xiàn)少量B細胞。 結(jié)論：在抗CD45RB誘導的免疫耐受中,B細胞可能介導了對T細胞各亞群比例的影響,且在中樞耐受中也起到一定作用。但單獨誘導B細胞的耐受只能延長移植物生存期,不能誘導完全耐受。B細胞缺失的情況下,抗CD45RB抗體對T細胞表面CD45RB表達下調(diào)更為快速,但與B細胞存在相比,最終CD3+CD45RBhiT細胞的比例是無明顯差別的。
[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.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R392

【參考文獻】

相關(guān)期刊論文 前1條

1 黃赤兵;張艮甫;金歡勝;范明齊;王平賢;賈維勝;馮嘉瑜;肖亞;方針強;;供體抗原特異性的CD4~+CD25~+Treg對腎移植大鼠體內(nèi)B細胞功能的影響[J];免疫學雜志;2009年01期

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本文編號：1896114