Okamoto Aoi, Shibuta Tatsuki, Morita Nanaka, Fujinuma Ryota, Shiraishi Masaya, Matsuda Reimi, Okada Mayu, Watanabe Satoe, Umemura Tsukuru, Takeuchi Hiroaki
Medical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, Japan.
Department of Medical Science Technology, School of Health Science at Fukuoka, International University of Health and Welfare, 137-1 Enokiz, Okawa 831-8501, Japan.
Microorganisms. 2025 Mar 26;13(4):753. doi: 10.3390/microorganisms13040753.
is a pathogenic bacterium that causes gastric and extragastric diseases. We have previously demonstrated that one of the mechanisms of -associated chronic immune thrombocytopenia involves immune complexes of platelets, a protein Lpp20 and an anti-Lpp20 antibody. However, it remains unclear how Lpp20 enters the body. We hypothesize that bacterial extracellular vesicles (bEVs) transport Lpp20. Thus, this study assessed Lpp20 in the bEVs released from seven clinical isolates, using immunoprecipitation (IP), immunoblotting (IB), and surface plasmon resonance imaging (SPRi), with anti-GroEL (a marker of bEVs) and anti-Lpp20 antibodies. Lpp20 and bEVs were each detected in lysates of all seven strains. IP-IB experiments demonstrated that bEVs containing Lpp20 were produced by five of the strains (J99, SS1, HPK5, JSHR3, and JSHR31). SPRi using an anti-Lpp20 antibody demonstrated significantly higher reflectance from the strain HPK5 than from its -disrupted strains ( < 0.01), indicating localization of Lpp20 on the bEVs' surface; Lpp20 may also be contained within bEVs. The bEVs containing Lpp20 were not detected from two clinical strains (26695 and JSHR6) or from two -disrupted strains (26695ΔLpp20 and HPK5ΔLpp20). Differences in Lpp20 detection in bEVs are likely due to variations in bEV production resulting from strain diversity.
是一种可导致胃部和胃外疾病的致病细菌。我们之前已经证明,与……相关的慢性免疫性血小板减少症的机制之一涉及血小板免疫复合物、一种Lpp20蛋白和一种抗Lpp20抗体。然而,Lpp20如何进入体内仍不清楚。我们推测细菌细胞外囊泡(bEVs)可转运Lpp20。因此,本研究使用免疫沉淀(IP)、免疫印迹(IB)和表面等离子体共振成像(SPRi),利用抗GroEL(bEVs的一种标志物)和抗Lpp20抗体,评估了从七种临床分离株释放的bEVs中的Lpp20。在所有七种菌株的裂解物中均检测到了Lpp20和bEVs。IP-IB实验表明,五种菌株(J99、SS1、HPK5、JSHR3和JSHR31)产生了含有Lpp20的bEVs。使用抗Lpp20抗体的SPRi显示,HPK5菌株的反射率明显高于其缺失菌株(<0.01),表明Lpp20定位于bEVs表面;Lpp20也可能包含在bEVs内。在两种临床菌株(26695和JSHR6)或两种缺失菌株(26695ΔLpp20和HPK5ΔLpp20)中未检测到含有Lpp20的bEVs。bEVs中Lpp20检测的差异可能是由于菌株多样性导致的bEV产生差异。