茶树菇凝集素检测醋酸格拉替雷诱导小鼠的CD8~＋T淋巴细胞表面糖基化研究

Detection of Surface Glycosylation of CD8^+T Lymphocytes in Glatiramer Acetate-treated Mice, Using Agrocybe aegerita Lectins

醋酸格拉替雷是治疗多发性硬化症的一线药物,可通过诱导调节性CD8+T细胞发挥免疫调节作用。为研究GA诱导的CD8+T淋巴细胞,我们将茶树菇分离的两种凝集素AAL(Agrocybe aegerita lectin)和AAL2(Agrocybe aegerita lectin 2),通过生物素标记结合流式细胞仪,分别分析GA诱导的免疫耐受小鼠模型中脾脏、腹股沟淋巴结CD8+T淋巴细胞的糖基化水平,及细胞因子的表达。发现脾脏AAL+CD8+,AAL2+CD8+T淋巴细胞的比例(%)分别为8.46±3.1和9.08±2.8,淋巴结AAL+CD8+,AAL2+CD8+T淋巴细胞的比例(%)分别为10.65±4.6和8.25±5.9,且脾脏、淋巴结CD8+T淋巴细胞中AAL+细胞的百分比均显著高于CD8-T淋巴细胞(p<0.05)。淋巴结中AAL+CD8+T细胞表达IFN-γ的细胞比例显著高于AAL-CD8+T细胞,AAL2+CD8+T细胞表达IFN-γ和perforin细胞比例均显著高于AAL2-CD8+T细胞。该结果为CD8+T淋巴细胞表面糖基化研究提供了初步数据,并暗示糖基化水平与细胞因子分泌有关。

Glatiramer acetate（GA） is a first-line agent for the treatment of multiple sclerosis, and it exerts an immunomodulatory function by inducing regulatory CD8＋ T cells. In order to investigate GA-induced CD8＋ T cells, the lectins AAL（Agrocybe aegerita lectin） and AAL2 were used to analyze the glycosylation level and cytokine expression in CD8＋ T lymphocytes in the spleen and inguinal lymph nodes in GA-induced, immune-tolerant mice, by biotin-labeling and flow cytometry. The results showed that the percentages of AAL＋CD8＋ and AAL2＋CD8＋ T lymphocytes in the spleen were 8.46 ± 3.1% and 9.08 ± 2.8%, respectively; and the percentages of AAL＋CD8＋ and AAL2＋CD8＋T lymphocytes in the inguinal lymph nodes were 10.65 ± 4.6% and 8.25 ± 5.9%, respectively. Moreover, the percentages of AAL＋ cells in CD8＋T lymphocytes in both the spleen and inguinal lymph nodes were significantly higher than those in the CD8- T lymphocytes（p 0.05）. In the inguinal lymph nodes, the proportion of AAL＋CD8＋ T cells expressing IFN-γ was significantly higher than that of AAL-CD8＋ T cells, and the proportion of cells expressing IFN-γ＋ and perforin in AAL2＋CD8＋ T cells was also significantly higher than that in AAL2-CD8＋ T cells. These studies provide preliminary data for studies on the surface glycosylation of CD8＋ T lymphocytes, and indicate that glycosylation level is associated with cytokine secretion.