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携带编码柔嫩艾美耳球虫抗原SO7基因的重组减毒鼠伤寒沙门氏菌疫苗株的免疫原性

Immunogenicity of recombinant attenuated Salmonella enterica serovar Typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen SO7.

作者信息

Konjufca Vjollca, Jenkins Mark, Wang Shifeng, Juarez-Rodriguez Maria Dolores, Curtiss Roy

机构信息

Biodesign Institute, Center for Infectious Diseases and Vaccinology, School of Life Sciences, Arizona State University, Tempe, Arizona 85287, USA.

出版信息

Infect Immun. 2008 Dec;76(12):5745-53. doi: 10.1128/IAI.00897-08. Epub 2008 Sep 22.

Abstract

Recombinant attenuated Salmonella vaccines against avian coccidiosis were developed to deliver Eimeria species antigens to the lymphoid tissues of chickens via the type 3 secretion system (T3SS) and the type 2 secretion system (T2SS) of Salmonella. For antigen delivery via the T3SS, the Eimeria tenella gene encoding sporozoite antigen SO7 was cloned downstream of the translocation domain of the Salmonella enterica serovar Typhimurium sopE gene in the parental pYA3868 and pYA3870 vectors to generate pYA4156 and pYA4157. Newly constructed T3SS vectors were introduced into host strain chi8879 (Delta phoP233 Delta sptP1033::xylE Delta asdA16), an attenuated derivative of the highly virulent UK-1 strain. The SopE-SO7 fusion protein was secreted by the T3SS of Salmonella. The vector pYA4184 was constructed for delivery of the SO7 antigen via the T2SS. The SO7 protein was toxic to Salmonella when larger amounts were synthesized; thus, the synthesis of this protein was placed under the control of the lacI repressor gene, whose expression in turn was dependent on the amount of available arabinose in the medium. The pYA4184 vector was introduced into host strain chi9242 (Delta phoP233 Delta asdA16 Delta araBAD23 Delta relA198::araC P(BAD) lacI TT [TT is the T4ipIII transcription terminator]). In addition to SO7, for immunization and challenge studies we used the EAMZ250 antigen of Eimeria acervulina, which was previously shown to confer partial protection against E. acervulina challenge when it was delivered via the T3SS. Immunization of chickens with a combination of the SO7 and EAMZ250 antigens delivered via the T3SS induced superior protection against challenge by E. acervulina. In contrast, chickens immunized with SO7 that was delivered via the T2SS of Salmonella were better protected from challenge by E. tenella.

摘要

针对鸡球虫病的重组减毒沙门氏菌疫苗被研发出来,用于通过沙门氏菌的3型分泌系统(T3SS)和2型分泌系统(T2SS)将艾美耳球虫属抗原递送至鸡的淋巴组织。为了通过T3SS递送抗原,将编码子孢子抗原SO7的柔嫩艾美耳球虫基因克隆到亲本pYA3868和pYA3870载体中鼠伤寒沙门氏菌血清型Typhimurium sopE基因的转运结构域下游,以产生pYA4156和pYA4157。新构建的T3SS载体被导入宿主菌株chi8879(Delta phoP233 Delta sptP1033::xylE Delta asdA16),它是高毒力UK-1菌株的减毒衍生物。SopE-SO7融合蛋白由沙门氏菌的T3SS分泌。构建了载体pYA4184用于通过T2SS递送SO7抗原。当合成大量SO7蛋白时,其对沙门氏菌有毒性;因此,该蛋白的合成置于lacI阻遏基因的控制之下,而lacI阻遏基因的表达又取决于培养基中阿拉伯糖的含量。将pYA4184载体导入宿主菌株chi9242(Delta phoP233 Delta asdA16 Delta araBAD23 Delta relA198::araC P(BAD) lacI TT [TT是T4ipIII转录终止子])。除了SO7,为了进行免疫和攻毒研究,我们使用了堆型艾美耳球虫的EAMZ250抗原,先前研究表明,当通过T3SS递送该抗原时,它能对堆型艾美耳球虫攻毒提供部分保护。用通过T3SS递送的SO7和EAMZ250抗原组合对鸡进行免疫,可诱导出对堆型艾美耳球虫攻毒的超强保护。相比之下,用通过沙门氏菌T2SS递送的SO7免疫的鸡,对柔嫩艾美耳球虫攻毒有更好的保护作用。

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