NKT细胞在小鼠皮肤移植口服免疫耐受中的作用

发布时间：2018-06-21 07:57

本文选题：NKT细胞 + 免疫耐受　；参考：《吉林大学》2011年硕士论文

【摘要】：目的：本实验探讨NKT细胞在小鼠皮肤移植模型中如何影响口服免疫耐受。目前由于免疫抑制剂高效性和临床广泛应用使同种异体移植进步显著,移植物存活时间延长,排斥率明显降低。但是长期以来,由于免疫抑制剂的毒副作用及排异反应,严重影响了移植物的长期存活和受者的生存质量。因此诱导特异性移植耐受,减少排斥反应,避免免疫抑制剂的毒副作用及解决移植器官长期存活成为焦点。大量实验表明,通过术前口服供体抗原可以诱导移植免疫耐受。NKT细胞是一类具有独特生物学活性的细胞亚群,在移植免疫耐受的形成中扮演着重要的角色。方法：建立同种异体小鼠尾-背皮肤移植模型,受鼠分别给予供体脾淋巴细胞灌胃,腹腔注射环孢素及供体脾淋巴细胞灌胃和环孢素腹腔注射联合应用,术后观察移植皮肤存活状况。应用流式细胞技术检测各组小鼠脾细胞中NKT细胞变化,并提取产生免疫耐受的小鼠的NKT细胞,过继性转移输注给正常受鼠,再行皮肤移植,观察移植皮肤存活时间,进一步研究NKT细胞诱导耐受情况。结果： (1)对照组小鼠移植皮片平均存活时间(MST)为12天,单纯口服组、单纯CSA组、口服+环孢素组的皮片存活时间分别为15天,14天和16.5天。与对照组比较,单纯CSA组皮片存活时间显著延长(P0.0001)；而与对照组比较,单纯口服组皮片存活时间也有显著延长(P0.0001),且诱导免疫耐受的效果优于单纯CSA组(P0.05)。口服+CSA组皮片移植物存活时间明显优于单纯口服组(P0.01)及单纯CSA组(P0.05)。 (2)流式细胞技术检测受鼠脾细胞中NKT细胞(NK1.1+CD3+)的百分比。与对照组比较,单纯CSA组脾细胞中NKT细胞百分比无显著变化,而单纯口服组,NKT细胞显著增加,P0.05；与单纯口服组比较,口服+CSA组NKT细胞百分比显著增加,P0.05；与对照组比较,NKT细胞百分比增加更为明显,P0.001。 (3)提取产生免疫耐受的小鼠的NKT细胞,并给予另一组小鼠鼠尾静脉过继转移输注,再行皮肤移植,移植皮肤较对照组的存活期明显延长(P0.001)。结论：通过口服供体抗原结合免疫抑制剂腹腔注射可以诱导小鼠同种异体皮肤免疫耐受,免疫耐受后受体脾细胞中NKT细胞明显增加,过继性转移输注耐受小鼠的NKT细胞可以诱导野生型小鼠的皮肤移植免疫耐受。
[Abstract]:Objective : To investigate the effect of NKT cells on oral immune tolerance in mouse skin transplantation model .

Methods : The rat tail - back skin transplantation model was established , and the mice were given donor spleen lymphocyte lavage , intraperitoneal injection of cyclosporin and donor spleen lymphocytes . The survival of transplanted skin was observed . The NKT cells were detected by flow cytometry . The NKT cells in the spleen cells of each group were detected by flow cytometry . The survival time of transplanted skin was observed and the tolerance of NKT cells was further studied .

Results :

( 1 ) The average survival time ( MST ) of the grafts in control group was 12 days . The survival time of the skin grafts was 15 days , 14 days and 16.5 days in the simple oral group and the simple CSA group . Compared with the control group , the survival time of the only CSA group was significantly prolonged ( P0.01 ) .
Compared with the control group , the survival time of the simple oral group was significantly prolonged ( P < 0.01 ) , and the effect of inducing immune tolerance was better than that of the simple CSA group ( P0.05 ) . The survival time of the oral + CSA group was obviously better than that of the simple oral group ( P0.01 ) and the simple CSA group ( P0.05 ) .

( 2 ) The percentage of NKT cells ( NK1 . 1 + CD3 + ) in spleen cells was detected by flow cytometry . Compared with the control group , the percentage of NKT cells in the spleen cells of the simple CSA group was not significantly changed , while only the NKT cells increased significantly in the simple oral group ( P0.05 ) .
Compared with the simple oral group , the percentage of NKT cells in oral and CSA group was significantly increased ( P0.05 ) .
Compared with the control group , the percentage of NKT cells increased more obviously , P0.001 .

( 3 ) The NKT cells were extracted from the mice with immune tolerance , and the other group of mice were given the adoptive transfer infusion of tail vein , then skin transplantation was performed , and the survival time of the transplanted skin was significantly prolonged compared with the control group ( P0.001 ) .

Conclusion : The immune tolerance of allogeneic skin can be induced by intraperitoneal injection of oral donor antigen and immune inhibitor , and the NKT cells in the spleen cells of the receptor after immune tolerance increase obviously , and the NKT cells in the adoptive transfer infusion tolerance mice can induce the immune tolerance of skin graft in wild type mice .
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.4

【参考文献】