{"title":"新弗朗西斯菌FTN_0096参与细胞内复制和宿主应答因子的鉴定","authors":"Dhandy Koesoemo Wardhana, Takashi Shimizu, Kenta Watanabe, Akihiko Uda, Masahisa Watarai","doi":"10.1371/journal.pone.0329626","DOIUrl":null,"url":null,"abstract":"<p><p>Francisella tularensis is the causative agent of the zoonotic disease tularemia. We investigated a pathogenic factor of F. tularensis subsp. novicida (F. novicida). Accordingly, we established a novel infection model using HeLa cells. F. novicida usually infects macrophage lineage cells and less frequently epithelial cells. We successfully infected HeLa cells expressing the Fc receptor (HeLa-FcγRII cells) using F. novicida supplemented with mouse serum containing F. novicida antibodies. A total of 2,232 transposon mutants of F. novicida were screened to determine the relatively fewer cytotoxic strains of the HeLa-FcγRII cells, and 13 strains were thus isolated. Sequencing analysis of transposon insertion sites identified 13 genes, including FTN_0096. We focused on FTN_0096. Although the F. novicida wild-type strain proliferated in HeLa-FcγRII and THP-1 cells, the number of intracellular FTN_0096 mutant decreased. FTN_0096 mutant cannot escape from phagolysosomes in the initial phases of infection. Moreover, FTN_0096 mutant was detected in the mitochondria and Golgi complex. These findings indicate the importance of FTN_0096 of F. novicida for intracellular replication in the cells.</p>","PeriodicalId":20189,"journal":{"name":"PLoS ONE","volume":"20 8","pages":"e0329626"},"PeriodicalIF":2.6000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12316277/pdf/","citationCount":"0","resultStr":"{\"title\":\"Identification of the Francisella novicida FTN_0096 as a factor involved in intracellular replication and host response.\",\"authors\":\"Dhandy Koesoemo Wardhana, Takashi Shimizu, Kenta Watanabe, Akihiko Uda, Masahisa Watarai\",\"doi\":\"10.1371/journal.pone.0329626\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Francisella tularensis is the causative agent of the zoonotic disease tularemia. We investigated a pathogenic factor of F. tularensis subsp. novicida (F. novicida). Accordingly, we established a novel infection model using HeLa cells. F. novicida usually infects macrophage lineage cells and less frequently epithelial cells. We successfully infected HeLa cells expressing the Fc receptor (HeLa-FcγRII cells) using F. novicida supplemented with mouse serum containing F. novicida antibodies. A total of 2,232 transposon mutants of F. novicida were screened to determine the relatively fewer cytotoxic strains of the HeLa-FcγRII cells, and 13 strains were thus isolated. Sequencing analysis of transposon insertion sites identified 13 genes, including FTN_0096. We focused on FTN_0096. Although the F. novicida wild-type strain proliferated in HeLa-FcγRII and THP-1 cells, the number of intracellular FTN_0096 mutant decreased. FTN_0096 mutant cannot escape from phagolysosomes in the initial phases of infection. Moreover, FTN_0096 mutant was detected in the mitochondria and Golgi complex. These findings indicate the importance of FTN_0096 of F. novicida for intracellular replication in the cells.</p>\",\"PeriodicalId\":20189,\"journal\":{\"name\":\"PLoS ONE\",\"volume\":\"20 8\",\"pages\":\"e0329626\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2025-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12316277/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"PLoS ONE\",\"FirstCategoryId\":\"103\",\"ListUrlMain\":\"https://doi.org/10.1371/journal.pone.0329626\",\"RegionNum\":3,\"RegionCategory\":\"综合性期刊\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q1\",\"JCRName\":\"MULTIDISCIPLINARY SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"PLoS ONE","FirstCategoryId":"103","ListUrlMain":"https://doi.org/10.1371/journal.pone.0329626","RegionNum":3,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"MULTIDISCIPLINARY SCIENCES","Score":null,"Total":0}
引用次数: 0
摘要
土拉菌是人畜共患疾病土拉菌病的病原体。我们研究了土拉菌亚种的一个致病因子。novicida的词性:因此,我们利用HeLa细胞建立了一种新的感染模型。新发梭菌通常感染巨噬细胞系细胞,很少感染上皮细胞。我们成功地用含新乳弧菌抗体的小鼠血清补充了表达Fc受体的HeLa细胞(HeLa-Fcγ - rii细胞)。筛选了2232个转座子突变体,确定了对hela - fc - γ - rii细胞毒性相对较小的菌株,并分离出13株。转座子插入位点测序鉴定出FTN_0096等13个基因。我们专注于FTN_0096。尽管野生型菌株在hela - fc - γ rii和THP-1细胞中增殖,但细胞内FTN_0096突变体数量减少。FTN_0096突变体在感染初期不能从吞噬溶酶体中逃脱。此外,在线粒体和高尔基复合体中检测到FTN_0096突变体。这些结果表明,FTN_0096基因在细胞内复制中起着重要的作用。
Identification of the Francisella novicida FTN_0096 as a factor involved in intracellular replication and host response.
Francisella tularensis is the causative agent of the zoonotic disease tularemia. We investigated a pathogenic factor of F. tularensis subsp. novicida (F. novicida). Accordingly, we established a novel infection model using HeLa cells. F. novicida usually infects macrophage lineage cells and less frequently epithelial cells. We successfully infected HeLa cells expressing the Fc receptor (HeLa-FcγRII cells) using F. novicida supplemented with mouse serum containing F. novicida antibodies. A total of 2,232 transposon mutants of F. novicida were screened to determine the relatively fewer cytotoxic strains of the HeLa-FcγRII cells, and 13 strains were thus isolated. Sequencing analysis of transposon insertion sites identified 13 genes, including FTN_0096. We focused on FTN_0096. Although the F. novicida wild-type strain proliferated in HeLa-FcγRII and THP-1 cells, the number of intracellular FTN_0096 mutant decreased. FTN_0096 mutant cannot escape from phagolysosomes in the initial phases of infection. Moreover, FTN_0096 mutant was detected in the mitochondria and Golgi complex. These findings indicate the importance of FTN_0096 of F. novicida for intracellular replication in the cells.
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