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为研究小鼠脾脏巨噬细胞(Mφ)对凋亡细胞诱导免疫应答的影响,我们用氯磷酸二钠脂质体(clodronate-liposomes,CL)清除小鼠脾脏Mφ,于不同时间点对三组(nave、PBSL、CL)小鼠选择性回输e450标记的DO11.10CD4~+T细胞、凋亡细胞及其相关抗原,然后用流式细胞仪检测小鼠血液与脾脏中免疫细胞的清除率、DO11.10CD4~+T细胞(即KJ126+T细胞)的增殖及Foxp3+Treg细胞的增殖。结果显示:(1)注射CL 1d后,外周血单核细胞的清除率约93%,脾脏中F4/80+Mφ、单核细胞与树突细胞的清除率分别为99%、90%、54%;(2)PBSL组与CL组中,KJ126+T细胞占CD4+T细胞的比率分别为1.32%、0.56%,差异有统计学意义(P<0.0001);Foxp3+T细胞占CD4~+T细胞的比率分别为0.69%、0.89%,差异有统计学意义(P=0.0004)。表明CL可以有效清除血液与脾脏中的免疫细胞;清除脾脏Mφ可以抑制KJ126+T细胞的增殖和促进Foxp3+Treg细胞的增殖。提示脾脏Mφ参与了提呈凋亡细胞相关抗原的过程,参与调节凋亡细胞诱导的抗原特异性T细胞的免疫应答能力及凋亡细胞诱导的免疫耐受。说明脾脏Mφ在凋亡细胞诱导的免疫反应中起着重要的作用。
In order to study the effect of Mφ on immune responses induced by apoptotic cells in mice, the Mφ of mouse spleen was removed by clodronate-liposomes (CL) (nave, PBSL, CL) mice e450-labeled DO11.10CD4 ~ + T cells, apoptotic cells and their associated antigen, and then detected by flow cytometry of immune cells in the blood and spleen of mice Clearance, proliferation of DO11.10CD4 ~ + T cells (ie KJ126 + T cells) and proliferation of Foxp3 + Treg cells. The results showed that: (1) The clearance rate of peripheral blood mononuclear cells was about 93% after CL 1d injection, and the clearance rates of F4 / 80 + Mφ, monocytes and dendritic cells in spleen were 99%, 90%, 54 %; (2) The ratio of KJ126 + T cells to CD4 + T cells in PBSL group and CL group was 1.32% and 0.56% respectively, with significant difference (P <0.0001); Foxp3 + T cells were 0.69%, 0.89%, the difference was statistically significant (P = 0.0004). It shows that CL can effectively eliminate immune cells in the blood and spleen. Clearing Mφ in the spleen can inhibit the proliferation of KJ126 + T cells and promote the proliferation of Foxp3 + Treg cells. These results suggest that Mφ is involved in the process of presenting apoptosis-related antigens and is involved in the regulation of immune response of antigen-specific T cells induced by apoptotic cells and immune tolerance induced by apoptotic cells. It shows that Mφ plays an important role in the immune response induced by apoptotic cells.