应用流式细胞术分析林麝外周血CD4^+、CD8^+T淋巴细胞

Analysis of CD4^+ and CD8^+ T Lymphocytes in Peripheral Blood of Forest Musk Deer(Moschus berezovskii) by Flow Cytometry

通过对圈养林麝(Moschus berezovskii)外周血淋巴细胞CD4^+、CD8^+亚群的检测,探讨林麝细胞免疫功能状态,并探索应用流式细胞仪分析其淋巴细胞亚群的方法,为研究林麝重大疾病的病理机制及诊断方法提供科学依据。本研究选取健康林麝和患呼吸道疾病林麝各5头,以双色流式细胞术检测其外周血淋巴细胞CD4^+、CD8^+亚群的含量,并进行比较。结果显示,羊源CD4、CD8的流式荧光抗体能够标记林麝细胞并有效检测;患病林麝与健康林麝相比,外周血CD4^+细胞含量无差异(P>0.05),CD8^+细胞含量则显著降低(P<0.01),CD4^+/CD8^+比值显著增高(P<0.01)。结果表明,患呼吸系统炎性疾病的林麝其外周血淋巴细胞CD8^+亚群变化显著,检测淋巴细胞亚群对林麝疾病的诊断有重要意义。

To investigate the immune functions of CD4^+ and CD8^+ lymphocyte subsets in peripheral blood of captive-breeding forest musk deer(Moschus berezovskii)and to explore the method for detecting lymphocyte subsets by flow cytometry will provide a scientific basis for studying the pathological mechanism and diagnostic methods of major diseases in this species.Five healthy musk deer and five musk deer with respiratory diseases were selected to compare CD4^+ and CD8^+ lymphocyte subgroups their peripheral blood by double-color flow cytometry.Data were processed and analyzed by SPSS18.0 software,and statistically described utilizing mean and standard deviation(Mean±SD).Differences between groups were analyzed by T-test,and P-value<0.05 was considered significant.The test results showed that flow-cytometric fluorescent antibodies against sheep-derived CD4 and CD8 could label the cells of forest musk deer effectively(Table 1 and Fig.1,2).Compared with healthy forest musk deer,the content of CD4^+cells in peripheral blood of diseased forest musk deer increased slightly,but there was no statistical difference(P>0.05)(Fig.3),while the content of CD8^+ cells decreased significantly(P<0.01)(Fig.3),and the ratio of CD4^+/CD8^+increased significantly(P<0.01)(Fig.3).The changes of CD8^+ lymphocyte subsets in peripheral blood of forest musk deer with respiratory inflammatory diseases were significant.Detection of lymphocyte subsets is of great importance in the diagnosis of forest musk deer diseases.