| 不同剂量流感病毒感染小鼠模型免疫应答的比较性研究 |
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| 引用本文： | 田晶,王宏杰,张轶博,隋天意.不同剂量流感病毒感染小鼠模型免疫应答的比较性研究[J].现代预防医学,2021,0(8):1473-1480. |
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| 作者姓名： | 田晶 王宏杰 张轶博 隋天意 |
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| 作者单位： | 1.锦州医科大学，辽宁 锦州 121000；2.大连医科大学附属第二医院，辽宁 大连 116027 |
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| 摘 要： | 目的 比较不同剂量流感病毒感染小鼠后肺组织急性损伤情况，初步探讨感染剂量与机体抗病毒的固有、适应性免疫应答的关系。方法 用流感病毒A/PR/8/34（PR8；H1N1）构建小鼠感染102 PFU和103 PFU剂量模型，HE法检测肺组织损伤情况，HA法和免疫荧光染色法检测肺组织病毒滴度和感染程度，RT - PCR法检测肺组织炎性因子干扰素α（interferon - α, IFN - α）、干扰素β（interferon - α, IFN - β）、白细胞介素1β（interleukin - 1β, IL - 1β）、白细胞介素6（interleukin - 6, IL - 6）、肿瘤坏死因子α（tumor necrosis factor - α, TNF - α）及Toll样受体7（Toll - like receptor - 7, TLR7）、髓样分化因子88（myeloid differentiation primary response gene 88, MyD88）、肿瘤坏死因子受体相关蛋白6（TNF receptor associated factor 6, TRAF6）和核因子κB（nuclear factor κB, NF - κB）mRNA水平，免疫组化法对肺组织TLR7、MyD88、TRAF6和NF - κB蛋白定位。流式细胞术检测肺组织、支气管肺泡灌洗液（bronchoalveolar lavage fluid, BALF）和淋巴结中CD4+、CD8+T细胞比例。结果 与102 PFU组比较，103 PFU组小鼠肺组织病毒滴度显著升高（t = 3.071，P = 0.008），炎症因子IFN - α、IFN - β、IL - 1β、IL - 6、TNF - α mRNA水平上升（t = 4.816，P = 0.003；t = 4.104，P = 0.006；t = 3.992，P = 0.007；t = 4.049，P = 0.007；t = 3.737，P = 0.009），TLR7、MyD88、TRAF6和NF - κB mRNA水平有不同程度的升高（t = 2.905，P = 0.027；t = 3.854，P = 0.008；t = 3.760，P = 0.009；t = 4.312，P = 0.005）及蛋白表达显著增加（t = 4.712，P = 0.003；t = 4.008，P = 0.007；t = 4.398，P = 0.005；t = 4.564，P = 0.004），BALF、肺组织和淋巴结中CD4+T细胞比例呈上升趋势（t = 3.771，P = 0.009；t = 2.463，P = 0.049；t = 3.386，P = 0.015），BALF和肺组织CD8+T细胞比例上调（t = 5.297，P = 0.002；t = 3.502，P = 0.013）。结论 病毒载量影响机体固有和适应性免疫应答状态，通过上调 TLR7 - MyD88 - TRAF6 - NF - κB信号通路促进下游炎症反应，并启动CD4+T和CD8+T细胞发挥抗病毒效应。
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| 关 键 词： | 不同剂量 流感病毒 炎症反应 免疫应答 |
Comparative study of immune responses in mice infected with influenza virus A/PR/8/34 (H1N1) at different doses |
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| TIAN Jing,WANG Hong-jie,ZHANG Yi-bo,SUI Tian-yi.Comparative study of immune responses in mice infected with influenza virus A/PR/8/34 (H1N1) at different doses[J].Modern Preventive Medicine,2021,0(8):1473-1480. |
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| Authors: | TIAN Jing WANG Hong-jie ZHANG Yi-bo SUI Tian-yi |
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| Affiliation: | *Jinzhou Medical University, Jinzhou, Liaoning 121000, China |
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| Abstract: | Abstract Objective To compare the acute lung injury of mice infected with different doses of influenza virus, and to preliminarily investigate the relationship between the infection dose and the innate and adaptive immune response to the virus. Methods The influenza model was constructed with influenza virus A/PR/8/34 (PR8; H1N1). We observed lung tissue damage of the mice under different infection doses of 102 PFU and 103 PFU. The lung virus titer and infection degree were detected by HA method and immunofluorescence staining method. RT-PCR method was used to detect levels of lung tissue inflammatory factors of IFN-α, IFN-β, IL-1β, IL-6, TNF-α and Toll-like receptor pathway related factors of TLR7, MyD88, TRAF6 and NF-κB mRNA. Immunohistochemical localization of TLR7, MyD88, TRAF6, and NF-κB proteins in lung tissue were performed. The percentages of CD4 + and CD8 +T cells in lung tissue, bronchoalveolar lavage fluid (BALF) and lymph nodes were measured by FCM. Results Compared to the 102 PFU group, the levels of pulmonary virus titer and inflammatory cytokines of IFN-α, IFN-β, IL-1β , IL-6, TNF-α mRNA were significantly increased in the 103 PFU group (t=3.071, P=0.008; t=4.816, P=0.003; t=4.104, P=0.006; t=3.992, P=0.007; t=4.049, P=0.007; t=3.737, P=0.009). The levels of mRNA and proteins of Toll-like receptor pathway related factors were increased to different degrees in the 103 PFU group compared to the 102 PFU group (t=2.905, P=0.027; t=3.854, P=0.008; t=3.760, P=0.009; t=4.312, P=0.005; t=4.712, P=0.003; t=4.008,P=0.007; t=4.398, P=0.005; t=4.564, P=0.004). The percentages of CD4+T cells in BALF, lung tissues and lymph nodes were upregulated (t=3.771, P=0.009; t=2.463, P=0.049; t=3.386, P=0.015). The percentages of CD8 +T cells in BALF and lung tissues showed an increasing trend (t=5.297, P=0.002; t=3.502, P=0.013). Conclusion Viral load affects the status of innate and adaptive immune response, promotes inflammation by up -regulating the TLR7 -MyD88 -TRAF6 -NF -κB signaling pathway, and initiates CD4+T and CD8+T cells to exert antiviral effects. |
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| Keywords: | Different doses Influenza virus Inflammatory response Immune response |
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