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为了了解体外抗体应答中T、B淋巴细胞的变化,以进一步阐明体外抗体应答的细胞学过程,并为完善体外免疫实验系统提供方向和依据,利用荧光激活细胞分选仪(FACS),在本实验室建立的人淋巴细胞体外免疫实验系统的基础上,对体外诱发抗体应答中淋巴细胞的变化进行了分析和研究,得到以下初步结果。一、T细胞所占比例的变化与设置培养物时细胞浓度相关,如果T细胞一开始占比例较大时,其变化为下降,否则其变化为上升。但最终都维持在30%上下。二、CD4~+(T辅助)细胞和CD8~+(T抑制)细胞的变化在扁桃体细胞和脾细胞中有所不同。在两周的培养过程中,前者无论加或不加C.M.(T细胞条件培液)或MTG(混合丝裂原),CD4~+细胞的比例均高于CD8~+细胞;而后者在加了C.M.或MTG后,CD8~+细胞的比例明显上升,但CD4~+细胞却有下降趋势。论文讨论了上述变化与免疫应答强度的可能关系。此外,亮氨酸甲酯(Leu-O-Me)的加入对CD8~+细胞没有影响。三、B淋巴细胞的变化,在培养两周过程中,无论扁桃体还是脾脏来源的细胞,B细胞富集的程度没有发生明显的变化,直至培养结束时,B细胞的比例仍维持在60%左右。T、B细胞间的比例也无明显变化,这表明所建立的体外免疫实验系统对B细胞的存活及其应答的诱发是适宜的。CD19~+B细胞的变化较为明显,其意义目前尚难推测。此外,一般认为CD5~+B细胞是一类分泌自身抗体的细胞,在培养过程中一直处于少数,未见增多。四、对于本实验中设置“框界”(“gating”)的合理性进行了讨论。
In order to understand the changes of T and B lymphocytes in vitro antibody response to further clarify the cytological process of antibody response in vitro and to provide the direction and basis for improving the in vitro immune system, fluorescence activated cell sorter (FACS) Laboratory established human lymphocytes in vitro immune experimental system based on the in vitro induced antibody response in lymphocyte changes were analyzed and the following preliminary results are obtained. First, the proportion of T cells and culture set changes in cell concentration, if the proportion of T cells at the beginning of a larger proportion, its decline, or change for the rise. But eventually maintained at 30% up and down. Second, CD4 ~ (T helper) cells and CD8 ~ + (T suppressor) changes in the cells of tonsils and spleen cells are different. In the two weeks of culture, the former with or without CM (T cell conditioned medium) or MTG (mixed mitogen), CD4 ~ + cells were higher than the proportion of CD8 ~ + cells; while the latter added After CM or MTG, the proportion of CD8 ~ + cells increased significantly, but CD4 ~ + cells showed a decreasing trend. The paper discusses the possible relationship between these changes and the intensity of immune responses. In addition, addition of Leu-O-Me had no effect on CD8 ~ + cells. Three, B lymphocyte changes, in the course of two weeks of culture, whether tonsil or spleen-derived cells, B cell enrichment levels did not change significantly until the end of the culture, the proportion of B cells remained at about 60% . T, B cells also showed no significant change in the ratio, indicating that the established in vitro immune experimental system of B cell survival and response induced induction is appropriate. The change of CD19 ~ + B cells is more obvious, its significance is hard to speculate at present. In addition, it is generally believed that CD5 ~ + B cells are a class of cells that secrete autoantibodies, and they have been in a minority during the culturing process without any increase. Fourth, the rationality of setting “gating” in this experiment was discussed.