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目的:研究在异基因骨髓细胞移植中葡萄球菌肠毒素B(SEB)诱导的耐受强度和细胞学特征。方法:选用C57BL/J小鼠作为受体,BALB/c小鼠作为供体。所有受体小鼠在接受6×107骨髓细胞前均接受6.0Gy60Coγ射线的照射,随机分成3组:组1为只接受6.0Gy60Coγ射线照射的对照组(照射对照组,RI);组2为照射后注射生理盐水(移植对照组,Tran.);组3为照射后注射60μgSEB(SEB组)。180d后由组2和组3实验鼠获得两组C57BL/L-BALB/c嵌合体小鼠。用流式细胞术分析移植后30~180d受体小鼠体内CD4+T、CD8+T、CD3+/NK1.1+NKT淋巴细胞亚群的数量和MHCH-2Kb、H-2Kd抗原表达的百分率,用MLR方法测定嵌合体小鼠淋巴细胞对ConA和异源性抗原的反应性。结果:(1)接受大剂量骨髓细胞移植后,注射SEB和注射生理盐水的两组小鼠均可存活180d以上,SEB组小鼠呈现出BALB/c供体小鼠的颜色特征(白色);Tran.组小鼠呈现出其灰白色。(2)SEB组嵌合体小鼠对ConA的反应性明显低于RI组和Tran.组;对异源性抗原的应答高于Tran.组。(3)SEB组小鼠外周血中CD4+T细胞的数量在移植后30~60d明显下降,随后增加;CD8+T细胞的数量不变。CD3+/NK1.1+NKT细胞的数量,从接受移植后30d开始增加,随着时间的延长而递增,到180d达到5.71%。存活180d的嵌合体小鼠,体内供体小鼠特有的MHCH-2Kd抗原的表达率高达80.95%,自体MHCH-2Kb抗原表达的百分率只占1.45%。(4)与SEB组相反,Tran.组小鼠CD8+T细胞的数量持续下降;CD3+/NK1.1+NKT细胞的数量的增加只在移植后180d上升达到5.07%。结论:在异基因骨髓细胞移植中,SEB诱导的耐受性比单纯骨髓移植诱导的耐受性强。SEB诱导的耐受性与移植早期特异性CD4+T细胞数量的减少和NKT细胞数的持续增加有关。反应性的降低表现为T细胞对ConA反应性的下降。
Objective: To study the intensity and cytological characteristics of staphylococcal enterotoxin B (SEB) induced in allogeneic bone marrow transplantation. Methods: C57BL / J mice were used as recipients and BALB / c mice as donor. All recipient mice received 6.0Gy60Coγ-ray irradiation before receiving 6 × 107 bone marrow cells, and were randomly divided into three groups: group 1 was a control group receiving only 6.0Gy60Coγ ray irradiation (irradiation control group, RI); group 2 was irradiation After injection of saline (transplantation control group, Tran.); Group 3 was irradiated with 60 μg of SEB (SEB group). Two groups of C57BL / L-BALB / c chimeric mice were obtained from group 2 and group 3 mice after 180 days. The numbers of CD4 + T, CD8 + T, CD3 + / NK1.1 + NKT lymphocyte subsets and the percentage of MH-2Kb and H-2Kd antigen expression in recipient mice after 30-180 d transplantation were analyzed by flow cytometry. Reactivity of Chimeric Mouse Lymphocytes to ConA and Heterologous Antigens by MLR Method. Results: (1) Both groups of mice injected with SEB and NS injected survived more than 180 days after receiving large doses of bone marrow cell transplantation, and the color characteristics (white) of BALB / c donor mice showed SEB mice. Tran. Mice appeared gray. (2) The response to ConA in SEB chimera mice was significantly lower than that in RI and Tran. The response to heterologous antigens was higher than that in Tran. (3) The number of CD4 + T cells in peripheral blood of SEB group decreased significantly from 30 to 60 days after transplantation, and then increased; the number of CD8 + T cells remained unchanged. The number of CD3 + / NK1.1 + NKT cells increased from 30 days after transplantation, and increased with time, reaching 5.71% after 180 days. The chimeric mice that survived for 180 days had 80.95% of MHCH-2Kd antigen specific for in vivo donor mice and only 1.45% of MHCH-2Kb antigen expressed in vivo. (4) In contrast to the SEB group, the number of CD8 + T cells in Tran. Mice continued to decline; the increase in the number of CD3 + / NK1.1 + NKT cells increased only 5.07% 180 days after transplantation. CONCLUSIONS: SEB-induced tolerance is more tolerant than allogeneic bone marrow transplantation in allogeneic bone marrow transplantation. The SEB-induced tolerance is associated with a reduction in the number of early-specific CD4 + T cells and a continual increase in the number of NKT cells. The decrease in reactivity is manifested by a decrease in the reactivity of T cells to ConA.