茶树菇凝集素AAL、AAL-2检测小鼠CD8+T淋巴细胞表面糖基化研究

Glycosylation of CD8+ T Lymphocytes from C57BL/6 Mice Recognized by Agrocybe aegerita Lectins

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。

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.