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Immunological and protective evaluation of purE/purK gene-deletion mutant of Brucella melitensis M5 strain

文献类型： 外文期刊

作者： Wang, Xin 1 ; Ji, Xue 1 ; Feng, Sheng 1 ; Sun, Yang 1 ; Zhu, Lingwei 1 ; Liu, Jun 1 ;

作者机构： 1.Chinese Acad Agr Sci, Changchun Vet Res Inst, Yujinxiang St 573, Changchun 130122, Jilin, Peoples R China

2.Jilin Univ, Inst Zoonosis, State Key Lab Diag & Treatment Severe Zoonot Infec, Coll Vet Med,Key Lab Zoonosis Res,Minist Educ, Changchun 130062, Jilin Province, Peoples R China

关键词： Brucella melitensis; Gene-deletion mutant; purE gene; purK gene; Vaccine

期刊名称：MICROBIAL PATHOGENESIS （ 影响因子：3.5； 五年影响因子：3.6 ）

ISSN： 0882-4010

年卷期： 2025 年 200 卷

页码：

收录情况： SCI

摘要： Brucellosis is listed by the World Health Organization as one of the seven most neglected global zoonotic diseases. A live-attenuated vaccine is still the main strategy used to prevent the spread of brucellosis. In this study, we constructed a two-gene (purE and purK)-deletion vaccine in Brucella melitensis vaccine strain M5 with homologous recombination. We evaluated its safety and immunogenicity in vitro and vivo, and its immunoprotective effect against B. melitensis wild-type strain 16M infection. In vitro, strain M5-Delta purEK induced 10.50 % lactose dehydrogenase (LDH) release, only half that induced by the vaccine strain M5 (29.92 % LDH). The ability of M5-Delta purEK to invade macrophages also decreased gradually, with a reduction of 1.02-log. In vivo, the hepatosplenic load and hepatosplenomegaly decreased significantly over time in mice inoculated with M5-Delta purEK (P < 0.05). It elicited anti-Brucella-specific IgG responses and triggered the secretion of gamma-interferon (IFN-gamma) along with a low level of interleukin-6 (IL-6). Serum samples inoculated with M5-Delta purEK were only 20 % positive on the Rose Bengal Plate Test. Hematoxylin-eosin staining showed only slight infiltration of neutrophils in the spleens of M5-Delta purEK-infected mice on days 14 and 42 after infection. The hepatocytes in the hepatic parenchyma were sparse and lightly stained cytoplasm, with few lymphocytes around the confluent area, which gradually returned to normal within 42 days. Like parental strain M5, strain M5-Delta purEK conferred protection against infection with B. melitensis wild-type strain 16M. In conclusion, we have shown that the deletion of purE and purK diminishes the cytotoxic effect of vaccine strain M5 on cells and organs, while leaving the degree of protection against infection with virulent wild-type Brucella strain 16M unchanged.

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