军事医学2026，Vol.50Issue(3)：182-191,10.DOI:10.7644/j.issn.1674-9960.2025-00289
一种人外周血单个核细胞来源淋巴样类器官优化模型的构建及其在重组破伤风毒素Hc片段蛋白免疫评价中的应用
Development and application of an optimized human PBMC-derived lymphoid organoid model for immune evaluation of recombinant TeNT-Hc protein
邵方泽 1张章 1廖伟年 1吕鹏 1于蕊 1徐俊杰 1张军1
作者信息
- 1. 军事科学院军事医学研究院前沿生物技术国家级重点实验室,北京 100071
- 折叠
摘要
Abstract
Objective To establish a lymphoid organoid model based on human peripheral blood mononuclear cells(PBMCs)and characterize the antigen-specific immune responses induced by the recombinant tetanus neurotoxin heavy chain(TeNT-Hc)protein based on this model.Methods Using viability and apoptosis of in vitro cultured PBMCs as indicators,the effects of key parameters,including different organoid culture methods,extracellular matrix,serum components and seeding densities,on the construction of lymphoid organoid model were compared.Recombinant TeNT-Hc protein was used as the model antigen to stimulate the organoids.Histological staining,flow cytometry and enzyme-linked immunosorbent assay(ELISA)were applied to evaluate the formation of germinal center-like structures,dynamic changes in specific immune cell subsets,and levels of antibody secretion.Results Under the optimized culture conditions,the cell viability of the constructed lymphoid organoids was 65.1%after 16 days of culture.Stimulation of lymphoid-like organoids with the model antigen resulted in marked activation and differentiation of B cell and T cell subsets,accompanied by the expansion of follicular helper T cell subsets.Germinal center-like structures were observed on day 8.Lymphoid organoids mounted B and T cell immune responses upon antigen stimulation,and antigen-specific antibody secretion was detected by day 6.PBMC-derived organoids from five healthy volunteers trended towards immune activation in response to antigen stimulation while retaining inter-individual differences in immune response intensity.Conclusion This established PBMC-derived lymphoid organoid model can effectively induce antigen-specific immune responses and possesses the dual ability to conduct standardized analysis and reflect the immune heterogeneity of the population.It provides a novel research tool for screening of vaccine candidate antigens and for the exploration of humoral immune mechanisms.关键词
淋巴样类器官/外周血单个核细胞/抗原特异性免疫应答/生发中心/体液免疫Key words
lymphoid organoids/peripheral blood mononuclear cells/antigen-specific immune responses/germinal center/humoral immunity分类
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邵方泽,张章,廖伟年,吕鹏,于蕊,徐俊杰,张军..一种人外周血单个核细胞来源淋巴样类器官优化模型的构建及其在重组破伤风毒素Hc片段蛋白免疫评价中的应用[J].军事医学,2026,50(3):182-191,10.
