{"title":"鲍曼不动杆菌对人宫颈癌上皮细胞的黏附性和细胞毒性:探索抗 OmpA 抗体的作用","authors":"Reyhaneh Rafiei Delfan , Zahra Fekrirad , Mohammadreza Jalali Nadoushan , Iraj Rasooli","doi":"10.1016/j.medmic.2024.100113","DOIUrl":null,"url":null,"abstract":"<div><div>We investigated the invasion of <em>Acinetobacter baumannii</em> strains to epithelial cells and elucidated the role of antibodies against outer membrane protein A (OmpA). <em>A. baumannii</em> ATCC 19606 and clinical isolate 58ST were utilized. OmpA was expressed, purified, and administered to BALB/c mice, inducing anti-OmpA antibodies. OmpA cytotoxicity was evaluated. Two <em>A. baumannii</em> strains were selected to infect human cervical HeLa cells. Serum resistance was determined at sera dilutions. Adhesion, internalization, and proliferation of live and killed <em>A. baumannii</em> in HeLa cells were examined with and without anti-OmpA sera. HeLa cell viability was assessed with and without exposure of live <em>A. baumannii</em> strains to anti-OmpA sera. Cytoskeleton inhibitor experiments were conducted on epithelial cells to probe microfilament and microtubule involvement in <em>A. baumannii</em> invasion. OmpA prompted antibody production without toxicity in mice. <em>A. baumannii</em> strains displayed varying cell invasion abilities, notably the clinical strain exhibiting the highest invasion. <em>A. baumannii</em> cells localized within vacuoles during internalization, migrating towards the nucleus, using a zipper-like invasion process. Bacterial proliferation within host cells led to HeLa cell death. Pre-treatment with anti-OmpA antibodies significantly curbed adhesion and invasion of <em>A. baumannii</em> in HeLa cells. Microscopic imaging provided proof of the intracellular presence of <em>A. baumannii</em> in HeLa cells. In conclusion, the OmpA plays a crucial part in <em>A. baumannii</em> - epithelial cell interactions. The results add to our knowledge of pathogenesis during the initial stages of infection by <em>A. baumannii</em>.</div></div>","PeriodicalId":36019,"journal":{"name":"Medicine in Microecology","volume":"22 ","pages":"Article 100113"},"PeriodicalIF":0.0000,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590097824000168/pdfft?md5=d4232009dc918160b6ff9afa917b2fec&pid=1-s2.0-S2590097824000168-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Adherence and cytotoxicity of Acinetobacter baumannii on human cervical carcinoma epithelial cells: Exploring the role of anti-OmpA antibodies\",\"authors\":\"Reyhaneh Rafiei Delfan , Zahra Fekrirad , Mohammadreza Jalali Nadoushan , Iraj Rasooli\",\"doi\":\"10.1016/j.medmic.2024.100113\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>We investigated the invasion of <em>Acinetobacter baumannii</em> strains to epithelial cells and elucidated the role of antibodies against outer membrane protein A (OmpA). <em>A. baumannii</em> ATCC 19606 and clinical isolate 58ST were utilized. OmpA was expressed, purified, and administered to BALB/c mice, inducing anti-OmpA antibodies. OmpA cytotoxicity was evaluated. Two <em>A. baumannii</em> strains were selected to infect human cervical HeLa cells. Serum resistance was determined at sera dilutions. Adhesion, internalization, and proliferation of live and killed <em>A. baumannii</em> in HeLa cells were examined with and without anti-OmpA sera. HeLa cell viability was assessed with and without exposure of live <em>A. baumannii</em> strains to anti-OmpA sera. Cytoskeleton inhibitor experiments were conducted on epithelial cells to probe microfilament and microtubule involvement in <em>A. baumannii</em> invasion. OmpA prompted antibody production without toxicity in mice. <em>A. baumannii</em> strains displayed varying cell invasion abilities, notably the clinical strain exhibiting the highest invasion. <em>A. baumannii</em> cells localized within vacuoles during internalization, migrating towards the nucleus, using a zipper-like invasion process. Bacterial proliferation within host cells led to HeLa cell death. Pre-treatment with anti-OmpA antibodies significantly curbed adhesion and invasion of <em>A. baumannii</em> in HeLa cells. Microscopic imaging provided proof of the intracellular presence of <em>A. baumannii</em> in HeLa cells. In conclusion, the OmpA plays a crucial part in <em>A. baumannii</em> - epithelial cell interactions. The results add to our knowledge of pathogenesis during the initial stages of infection by <em>A. baumannii</em>.</div></div>\",\"PeriodicalId\":36019,\"journal\":{\"name\":\"Medicine in Microecology\",\"volume\":\"22 \",\"pages\":\"Article 100113\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-09-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2590097824000168/pdfft?md5=d4232009dc918160b6ff9afa917b2fec&pid=1-s2.0-S2590097824000168-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Medicine in Microecology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2590097824000168\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Medicine in Microecology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2590097824000168","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
摘要
我们研究了鲍曼不动杆菌菌株对上皮细胞的侵袭,并阐明了外膜蛋白A(OmpA)抗体的作用。研究利用了鲍曼不动杆菌 ATCC 19606 和临床分离株 58ST。对 OmpA 进行表达、纯化并给 BALB/c 小鼠注射,诱导产生抗 OmpA 抗体。对 OmpA 的细胞毒性进行了评估。筛选出两种鲍曼不动杆菌菌株感染人类宫颈 HeLa 细胞。在血清稀释时测定血清抗性。在使用或不使用抗 OmpA 血清的情况下,检测 HeLa 细胞中活的和被杀死的鲍曼不动杆菌的粘附、内化和增殖情况。在鲍曼不动杆菌活菌株暴露于抗 OmpA 血清和未暴露于抗 OmpA 血清的情况下,评估 HeLa 细胞的存活率。对上皮细胞进行细胞骨架抑制剂实验,以探究微丝和微管在鲍曼不动杆菌侵袭中的参与情况。OmpA 能促使小鼠产生抗体,但无毒性。鲍曼不动杆菌菌株显示出不同的细胞侵袭能力,尤其是临床菌株显示出最高的侵袭能力。鲍曼不动杆菌细胞在内化过程中定位在空泡内,利用拉链式侵袭过程向细胞核迁移。细菌在宿主细胞内的增殖导致 HeLa 细胞死亡。用抗 OmpA 抗体进行预处理可显著抑制鲍曼不动杆菌在 HeLa 细胞中的粘附和入侵。显微成像证明了鲍曼不动杆菌在 HeLa 细胞内的存在。总之,OmpA 在鲍曼不动杆菌与上皮细胞的相互作用中起着至关重要的作用。这些结果增加了我们对鲍曼不动杆菌感染初期发病机制的了解。
Adherence and cytotoxicity of Acinetobacter baumannii on human cervical carcinoma epithelial cells: Exploring the role of anti-OmpA antibodies
We investigated the invasion of Acinetobacter baumannii strains to epithelial cells and elucidated the role of antibodies against outer membrane protein A (OmpA). A. baumannii ATCC 19606 and clinical isolate 58ST were utilized. OmpA was expressed, purified, and administered to BALB/c mice, inducing anti-OmpA antibodies. OmpA cytotoxicity was evaluated. Two A. baumannii strains were selected to infect human cervical HeLa cells. Serum resistance was determined at sera dilutions. Adhesion, internalization, and proliferation of live and killed A. baumannii in HeLa cells were examined with and without anti-OmpA sera. HeLa cell viability was assessed with and without exposure of live A. baumannii strains to anti-OmpA sera. Cytoskeleton inhibitor experiments were conducted on epithelial cells to probe microfilament and microtubule involvement in A. baumannii invasion. OmpA prompted antibody production without toxicity in mice. A. baumannii strains displayed varying cell invasion abilities, notably the clinical strain exhibiting the highest invasion. A. baumannii cells localized within vacuoles during internalization, migrating towards the nucleus, using a zipper-like invasion process. Bacterial proliferation within host cells led to HeLa cell death. Pre-treatment with anti-OmpA antibodies significantly curbed adhesion and invasion of A. baumannii in HeLa cells. Microscopic imaging provided proof of the intracellular presence of A. baumannii in HeLa cells. In conclusion, the OmpA plays a crucial part in A. baumannii - epithelial cell interactions. The results add to our knowledge of pathogenesis during the initial stages of infection by A. baumannii.