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目的:以NKT细胞形态和分化途径为焦点,研究超抗原SEB活化的NKT细胞亚群的特征。方法:C57BL/J小鼠脾细胞分别经SEB或者ConA诱导,收集体外扩增10d和5d的淋巴细胞以及获取正常黏附性巨噬细胞。用荧光抗体染色,流式细胞仪测定淋巴细胞和大淋巴细胞群表面CD69的表达、NKT细胞亚群百分数和不同NKT细胞亚群的分化途径。倒置显微镜400倍下比较大淋巴细胞的形态。结果:SEB活化的淋巴细胞和大淋巴细胞表面CD69分子表达由正常值的0.11%分别提高到55.00%和68.95%。大细胞群中CD8+和TCRVB8+NKT细胞亚群的百分数由原始的0.36%和0.81%分别提高到30.29%和31.48%。这些细胞位于流式细胞图的上部,是大体积细胞群。显微镜下显示SEB活化的大淋巴细胞体积不仅大于ConA活化的T淋巴细胞,而且比正常巨噬细胞大5倍以上,是非黏附性细胞。胞内质粒松散、胞质与核之比>1。SEB活化的CD8+和TCRVβ8+NKT细胞亚群均由T细胞直接分化而来。结论:SEB活化10d的NKT细胞亚群主要是CD8+NK1.1+和TCRVβ8+NK1.1+的NKT细胞。这些细胞是大体积淋巴细胞,应属于T细胞的亚群。
OBJECTIVE: To study the characteristics of NKT cell subsets activated by superantigen SEB with NKT cell morphology and differentiation pathway as the focus. Methods: The splenocytes of C57BL / J mice were induced by SEB or ConA respectively, and the lymphocytes were collected for 10d and 5d and the normal adherent macrophages were obtained. Fluorescent antibody staining was used to determine the expression of CD69 on the surface of lymphocytes and large lymphocytes, the percentage of NKT cells and the differentiation of different NKT subpopulations by flow cytometry. Inverted microscope 400 times larger lymphocyte morphology. Results: The expression of CD69 on the surface of activated lymphocytes and large lymphocytes increased from 0.11% to 55.00% and 68.95% respectively. The percentage of CD8 + and TCRVB8 + NKT cell subsets in the large cell population increased from 0.36% and 0.81% to 30.29% and 31.48% respectively. These cells are located in the upper part of the flow cytometry and are large populations of cells. Microscopically, SEB-activated macrophages were not only larger than ConA-activated T lymphocytes but were more than 5-fold larger than normal macrophages and were nonadherent cells. Intracellular plasmid loose, cytoplasm and nuclear ratio> 1. SEB-activated CD8 + and TCRVβ8 + NKT cell subsets are all differentiated directly by T cells. Conclusion: The NKT cell subsets activated by SEB for 10 days are mainly CD8 + NK1.1 + and TCRVβ8 + NK1.1 + NKT cells. These cells are bulky lymphocytes and should belong to a subset of T cells.