HSV-1突变株M6感染人支气管上皮细胞后对巨噬细胞介导的免疫反应的影响

张振潇; 张晶晶; 廖芸; 李丹丹; 李恒; 刘龙丁

doi:10.12259/j.issn.2095-610X.S20240702

HSV-1突变株M6感染人支气管上皮细胞后对巨噬细胞介导的免疫反应的影响

doi: 10.12259/j.issn.2095-610X.S20240702

张振潇^{1, 2},
张晶晶¹,
廖芸^{1, 2},
李丹丹^{1, 2},
李恒^{1, 2, ,},
刘龙丁^{1, 2, ,}

1.
中国医学科学院 & 北京协和医学院医学生物学研究所
2.
病毒疫苗研发系统创新研究重点实验室，云南昆明　650118

基金项目: 国家自然科学基金资助项目（82171817）；云南省自然科学基金资助项目（202301AS070066）

详细信息

作者简介:
张振潇（1999～），男，山东济南人，在读硕士研究生，主要从事疱疹病毒的感染与免疫研究工作

通讯作者:
李恒，E-mail：liheng@imbcams.com.cn

刘龙丁，E-mail：longdingl@gmail.com

中图分类号: R373.9
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出版历程
- 收稿日期: 2024-03-27
- 网络出版日期: 2024-06-14
- 刊出日期: 2024-07-25

Effects of HSV-1 Mutant Strain M6 on Macrophage-Mediated Immune Response after Infecting Human Bronchial Epithelial Cells

Zhenxiao ZHANG^{1, 2},
Jingjing ZHANG¹,
Yun LIAO^{1, 2},
Dandan LI^{1, 2},
Heng LI^{1, 2
, ,},
Longding LIU^{1, 2
, ,}

1.
Institute of Medical Biology，Chinese Academy of Medical Sciences & Peking Union Medical College，Kunming Yunnan 650118
2.
Key Laboratory of Systemic Innovative Research on Virus Vaccine，Chinese Academy of Medical Sciences，Kunming Yunnan 650118，China

摘要

摘要: 目的探究1型单纯疱疹病毒（herpes simplex virus type 1，HSV-1）突变株M6感染人支气管上皮细胞（16HBE细胞）后对巨噬细胞介导的免疫反应的影响。方法用HSV-1感染16HBE细胞分析培养液中细胞因子的变化；将巨噬细胞与被HSV-1毒株感染的16HBE细胞的上清液共培养并通过尾静脉回输至小鼠体内，分别在第1、3、7、28、56、90天对小鼠淋巴结细胞因子表达水平、脾脏T细胞比例变化、小鼠中和抗体表达水平以及特异性T细胞反应进行检测。结果 16HBE细胞被HSV-1突变株感染后，上清液中募集和激活巨噬细胞相关的细胞因子均较高水平表达但略低于野毒株组；尾静脉回输实验后，突变株组小鼠淋巴结炎症因子、趋化因子和T细胞的比例随时间发生了不同的变化，并引起了弱于野毒株组的体液免疫和强于野毒株组的特异性T细胞免疫反应，且仅极少数与野毒株组具有显著性差异（P < 0.05）。结论 16HBE细胞被HSV-1突变株M6感染后能够释放募集和激活巨噬细胞的细胞因子，使巨噬细胞携带HSV-1突变株的特异性活化信息，激活了宿主的免疫系统，诱导了宿主的体液免疫和细胞免疫。
- 1型单纯疱疹病毒 /
- 人支气管上皮细胞 /
- 巨噬细胞 /
- 突变株
Abstract: Objective To explore the impact of herpes simplex virus type 1 （HSV-1） mutant strain M6 on macrophage-mediated immune responses after infecting human bronchial epithelial cells （16HBE cells）. Methods HSV-1 infection of 16HBE cells was conducted to assess cytokine alterations in the culture medium; Macrophages were co-cultured with supernatants from HSV-1-infected 16HBE cells and subsequently reintroduced into mice via tail vein infusion. Cytokine expression levels in mouse lymph nodes, changes in spleen T cell proportions, mouse neutralizing antibody levels, and specific T cell responses were measured on days 1, 3, 7, 28, 56, and 90. Results Following infection of 16HBE cells with the HSV-1 mutant strain, cytokine expression in the supernatant, associated with macrophage recruitment and activation, was elevated, albeit slightly lower than that observed in the wild-type strain group. Following the tail vein infusion experiment, the mutant strain group exhibited temporal changes in the proportions of inflammatory factors, chemokines, and T cells within the mouse lymph nodes, resulting in weaker humoral immunity compared to the wild-type strain group, yet eliciting stronger specific T cell responses, with only a few parameters showing significant differences from the wild-type strain group （P < 0.05）. Conclusion 16HBE cells infected with the HSV-1 mutant strain M6 can release cytokines that recruit and activate macrophages, allowing macrophages to carry specific activation information of the HSV-1 mutant strain, activating the host's immune system and inducing the host's Humoral immunity and cellular immunity.
- Herpes simplex virus 1 /
- Human bronchial epithelial cells /
- Macrophages /
- Mutant strain

HTML全文

图 1 HSV-1 2种毒株感染16HBE细胞24 h后细胞因子表达水平的变化

2种毒株感染后均引起了各类细胞因子的不同变化。热图结果经过了归一化处理，具体方法是对数据采用了对数转换取其log10值，从而更有效地展示数据的变化范围和趋势。

Figure 1. Changes in cytokine expression levels after infection of 16HBE cells with two HSV-1 strains for 24 hours

下载: 全尺寸图片幻灯片

图 2 尾静脉回输实验后不同时间点小鼠的淋巴结中炎症因子的变化情况

A：淋巴结IL-6的变化情况；B：淋巴结GM-CSF的变化情况；C：淋巴结IL-1β、IL-10、TGF-β的变化情况；D：淋巴结TNF-α、IFN-γ的变化情况。^*P < 0.05，^**P < 0.01，^***P < 0.001，^****P < 0.0001。

Figure 2. Changes in inflammatory factors in lymph nodes of mice at different time points after tail vein reinfusion experiment

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图 3 尾静脉回输实验后不同时间点小鼠的淋巴结中趋化因子的变化情况及T细胞的分型情况

A：淋巴结主要趋化因子的变化情况；B：CD4⁺T细胞和CD8⁺T细胞的分型策略；C：淋巴结CD4⁺T细胞和CD8⁺T细胞的变化情况。^*P < 0.05，^**P < 0.01，^***P < 0.001，^****P < 0.0001。

Figure 3. Changes in chemokines and T cell typing in lymph nodes of mice at different time points after tail vein infusion experiment

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图 4 尾静脉回输实验后不同时间点小鼠血清的中和抗体水平的变化情况

A：中和抗体检测标准；B：血清中和抗体的变化情况。^*P < 0.05，^**P < 0.01，^***P < 0.001，^****P < 0.0001。

Figure 4. Changes in neutralizing antibody levels in mouse serum at different time points after the tail vein infusion experiment

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图 5 尾静脉回输实验后不同时间点小鼠脾脏细胞的IFN-γ、IL-4的ELISpot反应

A：小鼠脾脏细胞的IFN-γ的ELISpot反应；B：小鼠脾脏细胞的IL-4的ELISpot反应。^*P < 0.05，^**P < 0.01，^***P < 0.001，^****P < 0.0001。

Figure 5. ELISpot response of IFN-γ and IL-4 in mouse spleen cells at different time points after tail vein infusion experiment

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