感染伯氏疟原虫小鼠外周血淋巴细胞及其表面分子的检测  

Lymphocyte subsets and their surface molecules detection in the peripheral blood of Plasmodium berghei infected mouse

作　　者：郑贵星[1] 陈冰霞 张梦欣 金晨曦 谢世豪 齐艳伟 ZHENG Gui-xing;CHEN Bing-xia;ZHANG Meng-xin;JIN Chen-xi;XIE Shi-hao;QI Yan-wei(Department of Blood Transfusion,The First Affiliated Hospital of Guangzhou Medical University,Guangzhou 511436,China;The Third Clinical School,Guangzhou Medical University,Guangzhou511436,China;Department of Pathogenic Biology and Immunology,Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]广州医科大学附属第一医院输血科,广州511436 [2]广州医科大学第三临床学院,广州511436 [3]广州医科大学病原生物学与免疫学教研室,广州511436

出　　处：《现代免疫学》2021年第4期265-271,共7页Current Immunology

基　　金：国家自然科学基金青年科学基金(81902087);广东省自然科学基金(2017A030310535);广东省大学生创新创业训练计划项目(S201910570023,S201910570027);广州医科大学大学生创新创业训练项目(2018A036);广州医科大学第三临床学院大学生科研项目(2018A0024,2018A0020)。

摘　　要：为探讨C57BL/6小鼠在感染伯氏疟原虫(Plasmodium berghei)后外周血中的免疫细胞及其表面分子变化情况,将C57BL/6小鼠分为感染组和对照组(n=5),分别尾静脉注射伯氏疟原虫混悬液和生理盐水;感染第6天(Day 6,D6)处死小鼠,取外周全血并制备免疫细胞悬液;FACS检测小鼠B细胞、T细胞、CD8^(+)T细胞和NK细胞百分比及其表面分子CXCR3、CD69、CD62L表达水平变化情况。结果显示,疟原虫感染后,小鼠外周血B细胞百分比与对照组相比显著降低[(7.28±0.57)%vs(44.49±4.31)%,P<0.01],而T细胞、CD8^(+)T细胞和NK细胞百分比与对照组相比变化不显著[(33.26±6.65)%vs(35.99±3.45)%、(17.86±5.34)%vs(14.61±3.86)%、(13.32±1.92)%vs(11.75±1.64)%,均P>0.05];感染组B细胞和T细胞中CXCR3表达水平显著高于对照组[(10.52±0.65)%vs(5.46±1.59)%、(25.44±5.67)%vs(12.45±0.23)%,均P<0.05],而NK细胞中CXCR3表达相比对照组显著降低[(2.33±0.23)%vs(5.42±1.37)%,P<0.05],CD8^(+)T细胞中CXCR3表达水平高于对照组,但差异无统计学意义[(30.13±6.40)%vs(21.29±0.60)%,P>0.05];在感染组T细胞、CD8^(+)T细胞和NK细胞中CD69的表达水平显著高于对照组[(25.89±1.89)%vs(1.68±0.56)%、(20.43±3.91)%vs(2.34±1.05)%、(34.58±1.67)%vs(5.28±1.04)%,均P<0.001],而在B细胞中该指标高于对照组,但差异无统计学意义[(38.83±2.24)%vs(15.29±15.12)%,P>0.05];在感染组B细胞、T细胞、CD8^(+)T细胞和NK细胞中CD62L的表达水平相比对照组显著降低[(32.62±5.73)%vs(92.32±0.49)%、(2.60±0.50)%vs(76.33±8.34)%、(1.80±0.51)%vs(88.07±2.63)%、(0.25±0.06)%vs(14.69±2.37)%,均P<0.001]。综上,感染组小鼠外周血B细胞减少,B细胞和T细胞CXCR3表达水平升高,而NK细胞表达水平减少,T细胞、CD8^(+)T细胞和NK细胞高表达CD69,而感染组B细胞、T细胞、CD8^(+)T细胞和NK细胞低表达CD62L。这提示感染伯氏疟原虫后,小鼠的免疫应答处于低水平,可能机制是下调免疫细胞的CD62L表达水平。To investigate the changes of immune cells and their surface molecules in the peripheral blood of C57BL/6mice infected with Plasmodium berghei,mice were divided into the infected group and the normal group,and were injected Plasmodium berghei suspension or normal saline respectively through tail vein.The mice were sacrificed on Day 6,and the blood samples were taken to prepare peripheral blood immune cell suspension,the percentages of B cell,T cell,CD8^(+)T cell,NK cell and their surface molecules CXCR3,CD69and CD62Lwere measured by FACS.The results showed that,compared with the normal control,the B cell percentage was significantly reduced in the infected mice([7.28±0.57]%vs[44.49±4.31]%,P<0.01),while the percentages of T cell,CD8^(+)T cell and NK cell did not change significantly([33.26±6.65]%vs[35.99±3.45]%,[17.86±5.34]%vs[14.61±3.86]%,[13.32±1.92]%vs[11.75±1.64]%,all P>0.05);The expression level of CXCR3was significantly elevated in the B cells and T cells([10.52±0.65]%vs[5.46±1.59]%,[25.44±5.67]%vs[12.45±0.23]%,all P<0.05),but significantly decreased in NK cells in the infected group than in the normal group([2.33±0.23]%vs[5.42±1.37]%,P<0.05)and there was no significant change in CD8^(+)T cells([30.13±6.40]%vs[21.29±0.60]%,P>0.05);As to CD69level,it was significantly increased in the T cells,CD8^(+)T cells and NK cells in the infected group than those in the normal group([25.89±1.89]%vs[1.68±0.56]%,[20.43±3.91]%vs[2.34±1.05]%,[34.58±1.67]%vs[5.28±1.04]%,all P<0.01),the percentage of CD69+B cells was also elevated,but not statistically significant([38.83±2.24]%vs[15.29±15.12]%,P>0.05).The levels of CD62Lin the B cell,T cell,CD8^(+)T cell and NK cell were significantly decreased after the infection([32.62±5.73]%vs[92.31±0.49]%,[2.60±0.50]%vs[76.33±8.34]%,[1.80±0.51]%vs[88.07±2.63]%,[0.25±0.06]%vs[14.69±2.37]%,all P<0.001).In conclusion,in the infected group,B cell percentage decreased,the CXCR3levels increased in the B cell,T cell and decreased in the NK cell,CD69level increased

关 键 词：伯氏疟原虫 外周血 免疫细胞 细胞表面分子 

分 类 号：R392.5[医药卫生—免疫学]