Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens

作者全名："Zou, Jin-Tao; Jing, Hai-Ming; Yuan, Yue; Lei, Lang-Huan; Chen, Zhi-Fu; Gou, Qiang; Xiong, Qing-Shan; Zhang, Xiao-Li; Zhao, Zhuo; Zhang, Xiao-Kai; Zeng, Hao; Zou, Quan-Ming; Zhang, Jin-Yong"

作者地址："[Zou, Jin-Tao; Jing, Hai-Ming; Yuan, Yue; Lei, Lang-Huan; Chen, Zhi-Fu; Gou, Qiang; Xiong, Qing-Shan; Zhao, Zhuo; Zhang, Xiao-Kai; Zeng, Hao; Zou, Quan-Ming; Zhang, Jin-Yong] Third Mil Med Univ, Coll Pharm, Natl Engn Res Ctr Immunol Prod, Chongqing, Peoples R China; [Zou, Jin-Tao; Jing, Hai-Ming; Yuan, Yue; Lei, Lang-Huan; Chen, Zhi-Fu; Gou, Qiang; Xiong, Qing-Shan; Zhao, Zhuo; Zhang, Xiao-Kai; Zeng, Hao; Zou, Quan-Ming; Zhang, Jin-Yong] Third Mil Med Univ, Coll Pharm, Dept Microbiol & Biochem Pharm, Chongqing, Peoples R China; [Lei, Lang-Huan] Chongqing Med Univ, Childrens Hosp, Dept Crit Care Med, Chongqing, Peoples R China; [Zhang, Xiao-Li] Third Mil Med Univ, Coll Pharm, Dept Clin Hematol, Chongqing, Peoples R China"

通信作者："Zou, QM; Zhang, JY (corresponding author), Third Mil Med Univ, Coll Pharm, Natl Engn Res Ctr Immunol Prod, Chongqing, Peoples R China.; Zou, QM; Zhang, JY (corresponding author), Third Mil Med Univ, Coll Pharm, Dept Microbiol & Biochem Pharm, Chongqing, Peoples R China."

来源：PLOS PATHOGENS

ESI学科分类：MICROBIOLOGY

WOS号：WOS:000677728000002

JCR分区：Q1

影响因子：6.7

年份：2021

卷号：17

期号：7

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："Author summary Pore-forming toxins, a kind of exotoxins utilized by many pathogens as immune escaping weapons that targeting the immune cells and disturbing the immune system, are conventionally deemed as perfect antigens for vaccine development against infectious diseases. In this study, we reported that fusion of Hla(H35A), a typical pore-forming toxin toxoid, to candidate protein antigens from different species resulted in improved immunogenicity and protective efficacy. The improvement was mainly due to the increased antigen uptake and activating of immune-associated signaling pathways, probably by targeting ADAM10, the receptor of Hla on host immune cells. The importance of this work was to demonstrate the possible mechanisms of that pore-forming toxin function as atypical carrier protein to improve the immunogenicity of other proteins and confirm the potential of non-conservative but highly immunogenic exotoxins derived from hyper-virulent clinical strains for application in rational antigen design and vaccines development. Highly immunogenic exotoxins are used as carrier proteins because they efficiently improve the immunogenicity of polysaccharides. However, their efficiency with protein antigens remains unclear. In the current study, the candidate antigen PA0833 from Pseudomonas aeruginosa was fused to the alpha-hemolysin mutant Hla(H35A) from Staphylococcus aureus to form a Hla(H35A)-PA0833 fusion protein (HPF). Immunization with HPF resulted in increased PA0833-specific antibody titers, higher protective efficacy, and decreased bacterial burden and pro-inflammatory cytokine secretion compared with PA0833 immunization alone. Using fluorescently labeled antigens to track antigen uptake and delivery, we found that Hla(H35A) fusion significantly improved antigen uptake in injected muscles and antigen delivery to draining lymph nodes. Both in vivo and in vitro studies demonstrated that the increased antigen uptake after immunization with HPF was mainly due to monocyte- and macrophage-dependent macropinocytosis, which was probably the result of HPF binding to ADAM10, the Hla host receptor. Furthermore, a transcriptome analysis showed that several immune signaling pathways were activated by HPF, shedding light on the mechanism whereby Hla(H35A) fusion improves immunogenicity. Finally, the improvement in immunogenicity by Hla(H35A) fusion was also confirmed with two other antigens, GlnH from Klebsiella pneumoniae and the model antigen OVA, indicating that Hla(H35A) could serve as a universal carrier protein to improve the immunogenicity of protein antigens."

基金机构：National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31970138]

基金资助正文："This work was funded by the National Natural Science Foundation of China (grant number 31970138) to JYZ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."