[关键词]
[摘要]
本研究使用两种大型真菌茶树菇新型凝集素AAL(Agrocybe aegerita lectin)和AAL2(Agrocybe aegerita lectin 2),以检测小鼠CD8+ T淋巴细胞表面糖基化。用生物素标记的AAL和AAL2结合流式细胞仪,分别分析从C57BL/6小鼠脾脏和腹股沟淋巴结分离出来的CD8+T淋巴细胞表面的糖基化水平,及其细胞因子表达水平。结果发现脾脏中AAL+CD8+,AAL2+CD8+T淋巴细胞所占的比例(%)分别为6.72±3.00和12.18±5.28,而腹股沟淋巴结的AAL+CD8+,AAL2+CD8+T淋巴细胞所占的比例(%)分别为12.18±5.28和10.15±3.46。细胞因子表达水平结果发现小鼠脾脏AAL2+CD8+T细胞表达perforin的细胞比例低于AAL2-CD8+T细胞(p<0.05,有统计学意义),腹股沟淋巴结中AAL+CD8+T细胞表达perforin的细胞比例则高于AAL-CD8+T细胞(p<0.05,有统计学意义)。该数据为研究小鼠脾脏和腹股沟淋巴结CD8+ T细胞表面糖基化提供了新的数据,也为两种新型凝集素AAL和AAL2的应用提供理论依据。
[Key word]
[Abstract]
Two novel lectins, AAL (Agrocybe aegerita lectin) and AAL2 (Agrocybe aegerita lectin 2), were employed to detect the glycosylation of CD8+ T lymphocytes from C57BL/6 mouse spleen and inguinal lymph nodes. Biotinylated AAL and AAL2, combined with flow cytometry, were used to analyze the glycosylation of CD8+ T lymphocytes and cytokine expression. The proportion (%) of AAL+CD8+ and AAL2+CD8+ T lymphocytes in spleen was found to be 6.72±3.00 and 12.18±5.28, respectively. The proportion (%) of AAL+CD8+ and AAL2+CD8+ T lymphocytes in lymph nodes was 12.18±5.28 and 10.15±3.46, respectively. Moreover, the frequency of perforin+ cells in spleen AAL2+CD8+ T cells was lower than that in AAL2-CD8+ T cells (p<0.05), and the frequency of perforin+ cells in lymph node AAL+CD8+ T cells was higher than that in AAL-CD8+ T cells (p<0.05). This study provides new data on the glycosylation of spleen and inguinal lymph node CD8+ T cells and the application of two new lectins, AAL and AAL2.
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[基金项目]
国家自然科学基金项目(81102850);广东省卫生厅基金项目(A2011434);广东省教育厅育苗工程项目(LYM11070);东莞市科技局项目(2011108102049);湛江市科技攻关项目(2011C3109015);广东医学院大学生创新实验项目(2013ZYDC004)