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目的:检测GP96-肽复合物免疫小鼠脾淋巴细胞特定亚群CD8+细胞内的钙离子浓度变化,从而进一步探讨GP96-肽复合物疫苗的免疫效应和机制。方法:以GP96-肽复合物免疫小鼠的脾淋巴细胞作为研究对象, 用QUANTUM-RED标记的抗膜表面抗原CD8+进行膜表面染色的同时,进行FLUO-3/AM负载及细胞内CA2+浓度的检测,以双色流式细胞术检测特定亚群淋巴细胞内的CA2+浓度的变化,钙离子载体A23187、MNCL2和CON A作为小鼠脾淋巴细胞CD8+细胞[CA2+]I的刺激因素。结果:经GP96-肽复合物免疫小鼠脾淋巴细胞CD8+细胞[CA2+]I有一定水平的上调。与对照组相比,A23187和CON A可显著提高GP96-肽复合物免疫小鼠脾淋巴细胞CD8+细胞[CA2+]I 的水平。结论:[CA2+]I浓度的上升可能与GP96-肽复合物促进淋巴细胞CD8+T淋巴细胞的活化及增殖有关。
OBJECTIVE: To detect the changes of intracellular calcium concentration in specific subpopulation CD8 + cells of spleen lymphocytes immunized with GP96-peptide complex to further explore the immune effect and mechanism of GP96-peptide complex vaccine. Methods: The splenic lymphocytes of mice immunized with GP96-peptide complex were used as research objects. The membrane surface was stained with QUANTUM-RED-labeled anti-membrane surface antigen CD8 + and FLUO-3 / AM loaded with intracellular CA2 + The levels of CA2 + in specific subpopulations of lymphocytes were detected by two-color flow cytometry. Calcium ionophores A23187, MNCL2 and CON A were used as stimulation factors for the splenic lymphocyte CD8 + cells [CA2 +] I in mice. Results: The level of [CA2 +] I was up-regulated in splenic lymphocytes CD8 + cells immunized with GP96-peptide complex. Compared with the control group, A23187 and CON A significantly increased the level of [CA2 +] I in splenic lymphocytes CD8 + cells immunized with GP96-peptide complex. Conclusion: The increase of [CA2 +] I concentration may be related to the activation and proliferation of lymphocytes CD8 + T cells by GP96-peptide complex.