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
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.
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.