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微生物与感染  2023, Vol. 18 Issue (6): 321-330    DOI: 10.3969/j.issn.1673-6184.2023.06.001
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急慢性感染中B淋巴细胞应答规律比较研究的病毒工具构建
金程萌,白露,李雨檬,王倩瑜,李建华
复旦大学上海医学院基础医学院病原生物学系,教育部/卫健委/中国医科院医学分子病毒学重点实验室, 上海 200032
Construction of the viral tool for comparative study of B lymphocyte response patterns in acute and chronic infections
JIN Chengmeng, BAI Lu, LI Yumeng, WANG Qianyu, LI Jianhua
Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China
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摘要: 在急性或慢性等不同感染情况下B淋巴细胞应答规律各异。本研究旨在构建重组Armstrong株和重组Clone13株淋巴细胞脉络丛脑膜炎病毒(lymphocytic choriomeningitis virus, LCMV),以诱导针对WE株糖蛋白(glycoprotein, GP)特异的B细胞受体(B cell receptor, BCR)转基因细胞KL25HL B细胞应答,从而为比较急慢性感染过程中宿主B细胞的免疫应答差异建立实验模型。首先,通过共转染细胞,使之转录LCMV基因组片段并表达病毒蛋白,成功拯救出重组LCMV-Armstrong(recombinant LCMV-Armstrong,rARM)和重组LCMV-Clone13(recombinant LCMV-Clone13,rCL13)。测序分析表明,这2种重组病毒的GP序列被成功替换为LCMV-WE株的GP序列,并证实它们在感染细胞后具备蚀斑形成能力。随后,本研究使用rARM及rCL13感染野生型C56BL/6J小鼠。结果表明,这2种重组病毒保留了野生LCMV-Armstrong(wild type LCMV-Armstrong, wtARM)或野生LCMV-Clone13(wild type LCMV-Clone13, wtCL13)株等所具备的急性或慢性感染特性。具体而言,在rARM和rCL13建立感染后,rARM将在1周内被宿主免疫系统快速清除,而rCL13可在宿主体内持续存在超过2个月。最后,用不同LCMV感染过继转移了KL25HL B细胞的野生型小鼠。结果显示,与wtARM或wtCL13相比,感染rARM或rCL13后可明显诱导特异性B细胞发生激活、增殖等免疫应答反应。本研究构建的rARM及rCL13在感染小鼠并过继KL25HL B细胞后,可作为急慢性感染中B淋巴细胞应答规律比较研究的有力工具,对于深入理解体液免疫应答机制具有重要意义。
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金程萌
白露
李雨檬
王倩瑜
李建华
关键词 淋巴细胞脉络丛脑膜炎病毒B细胞感染模型    
Abstract:B lymphocyte response patterns vary in different infection scenarios, such as acute or chronic infections. This study aimed to construct recombinant Armstrong and Clone13 strains of lymphocytic choriomeningitis virus (LCMV) to induce the immune response of KL25HL B cells carrying the transgenic B cell receptor (BCR) specific to the glycoprotein (GP) of the WE strain, thereby establishing a model for comparing host B cell immune response differences between acute and chronic infection processes. Firstly, by co-transfecting cells with plasmids to transcribe LCMV genome fragments and express viral proteins, recombinant LCMV-Armstrong (rARM) and recombinant LCMV-Clone13 (rCL13) were successfully rescued. Sequencing analysis results indicated that the GP sequences of these two recombinant viruses were accurately replaced with the GP sequence of the LCMV-WE strain, and they were confirmed to have plaque-forming ability after infecting cells. For further characterizing the properties of rARM and rCL13, they were used to infect C57BL/6J mice. The results suggested that these two recombinant viruses retained the acute or chronic infection properties of wild type LCMV-Armstrong (wtARM) or wild type LCMV-Clone13 (wtCL13) strains. Specifically, after establishing infection, rARM would be rapidly cleared by the host immune system within a week, while rCL13 would persist in the host for more than two months. Finally, wild-type mice adoptively transferred with KL25HL B cells were infected with different LCMV strains to study the immune response. The results showed that, compared with wtARM or wtCL13, infection with rARM or rCL13 could significantly induce the activation and proliferation of KL25HL B cells. The rARM and rCL13 constructed in this study, after infecting mice that were adoptively transferred with KL25HL B cells, could serve as a powerful tool for comparative research on B lymphocyte response patterns in acute and chronic infections, which is of great significance for deepening understanding of humoral immune response mechanisms.
Key wordsLymphocytic choriomeningitis virus    B cells    Infection    Model
ZTFEL:  R392.3  
  R373  
基金资助:国家自然科学基金(81971937)
通讯作者: 李建华   
引用本文:   
金程萌,白露,李雨檬,王倩瑜,李建华. 急慢性感染中B淋巴细胞应答规律比较研究的病毒工具构建[J]. 微生物与感染, 2023, 18(6): 321-330.
JIN Chengmeng, BAI Lu, LI Yumeng, WANG Qianyu, LI Jianhua. Construction of the viral tool for comparative study of B lymphocyte response patterns in acute and chronic infections. JOURNAL OF MICROBES AND INFECTIONS, 2023, 18(6): 321-330.
链接本文:  
http://jmi.fudan.edu.cn/CN/10.3969/j.issn.1673-6184.2023.06.001     或     http://jmi.fudan.edu.cn/CN/Y2023/V18/I6/321

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