Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens.
Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens.
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DOI：
10.1371/journal.ppat.1009752
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发表时间：
2021-07
期刊：
影响因子：
6.7
通讯作者：
Zhang JY
中科院分区：
文献类型：
--
作者：
Zou JT;Jing HM;Yuan Y;Lei LH;Chen ZF;Gou Q;Xiong QS;Zhang XL;Zhao Z;Zhang XK;Zeng H;Zou QM;Zhang JY
高免疫原性外毒素被用作载体蛋白，因为它们有效地提高多糖的免疫原性。然而，它们对蛋白质抗原的效率仍不清楚。本研究将铜绿假单胞菌的候选抗原PA 0833与金黄色葡萄球菌的α-溶血素突变体HlaH 35 A融合，构建了HlaH 35 A-PA 0833融合蛋白（HPF）。与单独PA 0833免疫相比，用HPF免疫导致PA 0833特异性抗体滴度增加，保护效力更高，细菌负荷和促炎细胞因子分泌减少。使用荧光标记的抗原来跟踪抗原摄取和递送，我们发现HlaH 35 A融合显著改善了注射肌肉中的抗原摄取和向引流淋巴结的抗原递送。体内和体外研究均表明，HPF免疫后增加的抗原摄取主要是由于单核细胞和巨噬细胞依赖的巨胞饮作用，这可能是HPF与Hla宿主受体ADAM 10结合的结果。此外，转录组分析表明，几种免疫信号通路被HPF激活，揭示了HlaH 35 A融合提高免疫原性的机制。最后，HlaH 35 A融合蛋白的免疫原性也得到了其他两种抗原，肺炎克雷伯氏菌GlnH和模型抗原OVA的证实，表明HlaH 35 A可以作为一种通用的载体蛋白，以提高蛋白抗原的免疫原性。孔形成毒素是一类被许多病原体利用的免疫逃逸武器，它靶向免疫细胞，干扰免疫系统，被认为是开发抗感染性疾病疫苗的理想抗原。在这项研究中，我们报告了融合HlaH 35 A，一个典型的孔形成毒素类毒素，从不同物种的候选蛋白抗原导致提高免疫原性和保护效力。这种改善主要是由于抗原摄取增加和免疫相关信号通路的激活，这可能是通过靶向宿主免疫细胞上Hla的受体ADAM 10实现的。本工作的重要性在于证明孔形成毒素作为非典型载体蛋白的可能机制，以提高其他蛋白的免疫原性，并确认来自高毒力临床菌株的非保守但高免疫原性外毒素用于合理抗原设计和疫苗开发的潜力。
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 α-hemolysin mutant HlaH35A from Staphylococcus aureus to form a HlaH35A-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 HlaH35A 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 HlaH35A fusion improves immunogenicity. Finally, the improvement in immunogenicity by HlaH35A fusion was also confirmed with two other antigens, GlnH from Klebsiella pneumoniae and the model antigen OVA, indicating that HlaH35A could serve as a universal carrier protein to improve the immunogenicity of protein antigens. 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 HlaH35A, 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.
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DOI：
10.1084/jem.20091990
发表时间：
2010-03-15
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Gibb DR;El Shikh M;Kang DJ;Rowe WJ;El Sayed R;Cichy J;Yagita H;Tew JG;Dempsey PJ;Crawford HC;Conrad DH
通讯作者：
Conrad DH
影响因子：
7.3
作者：
Bianconi, Irene;Alcala-Franco, Beatriz;Bragonzi, Alessandra
通讯作者：
Bragonzi, Alessandra
DOI：
10.1126/science.1219364
发表时间：
2012-06-01
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
King NP;Sheffler W;Sawaya MR;Vollmar BS;Sumida JP;André I;Gonen T;Yeates TO;Baker D
通讯作者：
Baker D
影响因子：
17.1
作者：
Liang, Frank;Lindgren, Gustaf;Lore, Karin
通讯作者：
Lore, Karin
影响因子：
6.4
作者：
Kebaier, Chahnaz;Chamberland, Robin R.;Duncan, Joseph A.
通讯作者：
Duncan, Joseph A.