伯氏疟原虫ANKA株感染小鼠的T细胞、NK细胞及细胞因子变化

doi:10.11843/j.issn.0366-6964.2022.11.026

摘要/Abstract

摘要： 旨在系统分析伯氏疟原虫感染引起宿主T细胞、NK细胞、Tim-3表达及细胞因子的变化。选取64只雌性C57BL/6小鼠随机分为8组，每组8只，分别于感染后0、4、7、9、11、13、16和19 d获取小鼠脾及外周血免疫细胞，利用流式细胞术检测小鼠主要免疫细胞亚群及免疫检查点分子Tim-3表达水平的变化；同时检测血清中细胞因子的变化。结果表明，感染疟原虫后，小鼠脾CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及NK细胞的比例均逐渐降低（P<0.01），且伴随着3种细胞Tim-3表达量的升高（P<0.01）。小鼠外周血CD3⁺CD4⁺ T细胞的比例呈先降低后升高趋势，CD3⁺CD8⁺ T细胞的比例呈先升高后降低趋势（P<0.05）；小鼠外周血CD3^-NK1.1⁺细胞的比例呈先降低后升高趋势，但感染末期，其比例仍低于未感染组（P<0.05）。Tim-3分子在外周血CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及CD3^-NK1.1⁺细胞的表达均显著升高（P<0.05）。感染疟原虫后，小鼠血清中促炎细胞因子IL-2的分泌量均显著高于未感染组（P<0.05）；促炎细胞因子IFN-γ、TNF-α和IL-6在血清中分泌均呈先升高后降低趋势（P<0.05）；具有免疫抑制作用的细胞因子IL-10呈逐渐升高趋势，且感染后期急剧升高（P<0.001）。以上结果表明，感染疟原虫后，小鼠的特异性免疫反应发挥了一定的杀伤作用，但由于Tim-3免疫检查点分子及一些发挥免疫抑制作用的细胞因子（IL-10）的过度表达，有利于疟原虫逃避宿主的免疫捕杀作用。该研究提示了从宿主免疫抑制角度研究疟原虫感染的重要性。

关键词: 伯氏疟原虫, T细胞, NK细胞, Tim-3, 细胞因子

Abstract: This study aims to systematically analyze the changes of hosts' T cells, NK cells, expression of Tim-3 and cytokines post infection with Plasmodium berghei (P. berghei) in mice. A total of 64 female C57BL/6 mice were selected and randomly divided into 8 groups, with 8 in each group. Mice spleen and peripheral blood immune cells were obtained on 0, 4, 7, 9, 11, 13, 16 and 19 days post infection, and flow cytometry was used to detect the changes of main immune cell subsets and the expression of immune checkpoint molecule Tim-3. Mice sera were collected to detect the changes in cytokine levels. Results showed that the proportions of CD3⁺CD4⁺ T cells, CD3⁺CD8⁺ T cells and NK cells in mice spleen were gradually reduced post infection with P. berghei (P<0.01), and this was accompanied by an increase in the expression of Tim-3 on the surface of these three types of cells (P<0.01). In mice peripheral blood, the proportion of CD3⁺CD4⁺ T cells decreased first and then increased, proportion of CD3⁺CD8⁺ T cells increased first and then decreased (P<0.05). The proportion of circulatory CD3^-NK1.1⁺ cells also decreased first and then increased, but at the end of infection time, its proportion was still lower than that of the uninfected group. The expression level of Tim-3 in peripheral blood of CD3⁺CD4⁺ T cells, CD3⁺CD8⁺ T cells and CD3^-NK1.1⁺ cells all increased significantly (P<0.05). Post infection with P. berghei, the secretion of pro-inflammatory cytokine IL-2 in mice serum was significantly higher than that in the uninfected group (P<0.05); The secretion of pro-inflammatory cytokine IFN-γ, TNF-α and IL-6 in mice sera all increased first and then decreased, while the immunosuppressive cytokine IL-10 showed a gradual increase trend, and increased sharply in the later stage of infection (P<0.001). The results of this study showed that post infection with Plasmodium, the specific immune response of mice played a certain killing effect, but due to the overexpression of immune checkpoint molecule (Tim-3 etc.) and some immunosuppressive cytokines (IL-10 etc.), which helps the malaria parasite to escape from the host's immune killing process. This work proposes the importance of studying Plasmodium infection from the perspective of host immunosuppression.

Key words: Plasmodium berghei, T cell, NK cell, Tim-3, cytokines

中图分类号:

S852.72

张义伟, 苏紫薇, 李其龙, 陈冉, 姜宁. 伯氏疟原虫ANKA株感染小鼠的T细胞、NK细胞及细胞因子变化[J]. 畜牧兽医学报, 2022, 53(11): 4008-4018.

ZHANG Yiwei, SU Ziwei, LI Qilong, CHEN Ran, JIANG Ning. The Changes of T Cells, NK Cells and Cytokines in Mice Infected with Plasmodium berghei ANKA Strain[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4008-4018.

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