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巨噬细胞(macrophages,Mφ)在机体正常发育、体内平衡、组织修复和对病原体的免疫反应中起着重要作用。为了研究豚鼠(Cavia porcellus)Mφ在结核免疫中的作用及机制,本研究通过淀粉肉汤溶液预刺激的方法分离培养了豚鼠腹腔Mφ,并进行了细胞形态学观察、细胞活力、吞噬功能、非特异性酯酶染色和细胞表面特异性抗原表达检测等一系列鉴定。在此基础上,利用结核分枝杆菌(Mycobacterium tuberculosis,MTB)卡介苗(Bacillus Calmette-Guérin,BCG)感染分离培养的Mφ,并分析BCG对MφNO的产生和细胞凋亡的影响。结果表明,分离培养的腹腔Mφ具有Mφ所特有的形态特征、较强细胞活力和吞噬能力,并可检测到特异性的白细胞分化抗原14(cluster of differentiation 14,CD14)、CD40和CD68等的表达。与未感染的对照组相比,BCG感染24 h后Mφ产生的NO极显著增加(P<0.01),细胞的凋亡率也极显著升高(P<0.01)。表明淀粉肉汤溶液刺激的方法可成功获取豚鼠腹腔Mφ,同时证实BCG感染后能诱导其产生的NO增多,并且细胞凋亡率增加,提示其可能在抗结核免疫中起重要作用。研究结果为MTB和Mφ相互作用研究提供细胞模型和数据支撑。
Macrophages (Mφ) play an important role in normal body development, homeostasis, tissue repair and immune responses to pathogens. In order to study the role and mechanism of Mφ in the immunization of MΦ in guinea pigs, in this study, guinea pig peritoneal Mφ was isolated and cultured by pre-stimulation with starch broth solution. Cell morphology, cell viability, phagocytosis, Heterosexual esterase staining and cell surface antigen-specific detection and a series of identification. On this basis, Mφ was isolated from Mycobacterium tuberculosis (MTB) -treated Bacillus Calmette-Guérin (BCG) and the effect of BCG on MφNO production and apoptosis was analyzed. The results showed that Mφ isolated from the peritoneal cavity possessed the characteristic features of Mφ, strong cell viability and phagocytosis, and detected the expression of cluster of differentiation 14 (CD14), CD40 and CD68 . Compared with uninfected control group, the NO produced by Mφ significantly increased (P <0.01) and the apoptosis rate significantly increased (P <0.01) 24 h after BCG infection. The results showed that the stimulation of starch broth solution could successfully obtain Mφ in guinea pig peritoneal cavity, and at the same time confirmed that the increase of NO induced by BCG infection and the increase of apoptosis rate suggested that it may play an important role in anti-TB immunity. The results provide a cellular model and data support for the MTB and Mφ interaction studies.