gp96肽复合物免疫脾巨噬细胞对细胞内钙离子动态变化及形态学观察

目的：通过研究gp96-肽复合物免疫小鼠脾巨噬细胞细胞内Ca^2 的变化及其形态学的改变，进一步探讨肿瘤细胞中提纯的热休克蛋白gp96-肽复合物的抗肿瘤免疫作用和机制。方法：以gp96-肽复合物免疫的小鼠脾巨噬细胞作为研究对象，用Fluo-3／AM负载细胞．激光扫描共聚焦显微镜检测其细胞内的Ca^2 浓度及观察Con A刺激对细胞内Ca^2 影响；扫描电镜观察细胞的形态学变化。结果：与对照组相比。经gp96-肽复合物致敏的小鼠脾巨噬细胞内的Ca^2 浓度显著增加；其细胞内的Ca^2 变化对Con A刺激更加敏感；免疫后的小鼠脾巨噬细胞呈典型的激活状态。结论：gp96-肽复合物可在体内激活脾巨噬细胞；活化的脾巨噬细胞在Con A刺激时Ca^2 浓度的变化敏感。gp96-肽复合物的抗肿瘤免疫过程中促进小鼠脾巨噬细胞活化起着重要的作用。

Intercellular Ca2+ and morphology changes on gp96-peptide complexes immunized mouse spleen macrophages

Objective:To further investigate the anti-tumor effect and mechanism of Heat Shock Protein gp96-peptide complexes derived from malignant cells by observing the intercellular Ca 2 changes and morphology changes of immuno-activated spleen macrophages.Methods:The spleen macrophages from mice immunized with gp96-peptide complexes were used as research target.The cell was stained with highly sensitive Ca 2 fluorescent indicator Fluo-3/AM. The intercellular calcium concentration and Con A-stimulated intercellular Ca 2 changes were measured by laser scanning confocal microscope (LSCM).The morphology changes of macrophages were observed by scanning electron microscope (SEM).Results:Compared with the corresponding control group,the intercellular calcium concentration of immuno-activated spleen macrophages is largely increased; ConA-stimulated intercellular Ca 2 changes of immuno-activated spleen macrophages are more significant; the macrophages are in active state after immunization.Conclusions:gp96-peptide complexes may activate the spleen macrophages of mice in vitro.The actived cells are sensitive to ConA stimulation.It is suggested that gp96-peptide complexes play an important role in activating spleen macrophages.