Norman et al Clostridioides difficile Toxin B subverts germinal center and antibody recall responses by stimulating a drug-treatable CXCR4-dependent mechanism

Raw data for the Cell Reports manuscript concerning impact of TcdB2 on germinal center reactions. The PDF attachment included (Norman et al project metadata 110623) lists all information needed to analyze and interpret the raw data and is broken down figure by figure corresponding to its organization in the manuscript. On 04/04/24, this entry was amended to add a PDF of the project metadata corresponding to the manuscript revisions (Norman et al project metadata ADDENDUM 040424) and to add raw data corresponding to new experiments that were added during the course of revision. On 04/26/24 following provisional acceptance of the revised manuscript, the final dataset was re-uploaded. PROJECT SUMMARY: Recurrent Clostridioides difficile infection (CDI) results in significant morbidity and mortality. We previously established that CDI in mice did not protect against reinfection and was associated with poor pathogen-specific B cell memory (Bmem), recapitulating our observations with human Bmem. Herein, we demonstrate that the secreted toxin TcdB2 is responsible for subversion of Bmem responses. TcdB2 from an endemic C. difficile strain delayed IgG class switch following vaccination, attenuated IgG recall to a vaccine booster, and prevented germinal center formation. The mechanism of TcdB2 action included increased B cell CXCR4 expression and responsiveness to its ligand CXCL12, accounting for altered cell migration and a failure of germinal center-dependent Bmem. These results were reproduced in a C. difficile infection model and an FDA-approved CXCR4-blocking drug rescued germinal center formation. We therefore provide mechanistic insights into C. difficile-associated pathogenesis and illuminate a target for clinical intervention to limit recurrent disease.

本数据集为《Cell Reports》期刊投稿稿件的原始数据，主题为TcdB2对生发中心（germinal center）反应的影响。随附的PDF文件（Norman等项目元数据 110623）涵盖了分析与解读该原始数据所需的全部信息，并依照稿件的排版结构分图梳理对应内容。2024年4月4日，本条目完成更新：新增了对应稿件修订版的项目元数据PDF（Norman等项目元数据 补编 040424），并补充了修订阶段新增实验的原始数据。2024年4月26日，在修订稿件获期刊临时接收后，最终数据集已重新上传。

项目摘要：复发性艰难梭菌感染（Recurrent Clostridioides difficile infection, CDI）可导致严重的发病与死亡风险。本团队前期研究证实，小鼠感染艰难梭菌后无法抵御再次感染，且伴随病原体特异性B细胞记忆（Bmem）缺陷，该结果与我们在人类Bmem中观察到的现象一致。本研究表明，地方性流行艰难梭菌菌株分泌的毒素TcdB2是扰乱Bmem应答的核心因子。TcdB2可延缓疫苗接种后的IgG类别转换，削弱加强疫苗接种后的IgG回忆应答，并阻断生发中心形成。TcdB2的作用机制包括上调B细胞表面趋化因子受体4（CXCR4）的表达水平，增强其配体趋化因子配体12（CXCL12）的应答能力，进而改变细胞迁移模式，导致生发中心依赖型Bmem生成失败。上述实验结果在艰难梭菌感染模型中得到了重复验证，且经美国食品药品监督管理局（FDA）批准的CXCR4阻断剂可挽救生发中心的形成。综上，本研究阐明了艰难梭菌相关致病机制，并为限制复发性感染的临床干预提供了潜在靶点。