PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice

BackgroundSalmonellosis caused by Salmonella sp. is a foodborne zoonotic disease that poses a significant threat to public health security. Vaccination is a safe and effective strategy for preventing and controlling Salmonella infections. PipC is a chaperone protein associated with Salmonella invasi...

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Main Authors: Lu Zhang, Yubin Chen, Zhigang Yan, Yuntai Li, Xiaowen Yang, Li Chen, Yanying Zhang, Yingyu Chen, Yonghui Li, Qiumei Shi, Tonglei Wu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1631008/full
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author Lu Zhang
Yubin Chen
Zhigang Yan
Yuntai Li
Xiaowen Yang
Li Chen
Yanying Zhang
Yingyu Chen
Yonghui Li
Qiumei Shi
Tonglei Wu
author_facet Lu Zhang
Yubin Chen
Zhigang Yan
Yuntai Li
Xiaowen Yang
Li Chen
Yanying Zhang
Yingyu Chen
Yonghui Li
Qiumei Shi
Tonglei Wu
author_sort Lu Zhang
collection DOAJ
description BackgroundSalmonellosis caused by Salmonella sp. is a foodborne zoonotic disease that poses a significant threat to public health security. Vaccination is a safe and effective strategy for preventing and controlling Salmonella infections. PipC is a chaperone protein associated with Salmonella invasion proteins which is crucial for bacteria to invade host cells.MethodsIn this study, a ΔpipC mutant strain was generated. Subsequently, we examined the environmental stress tolerance of the mutant strain through in vitro simulation experiments. Moreover, its virulence by employing cell and mouse infection models was investigated. Furthermore, we utilized a mouse model to further explore its potential as an attenuated live vaccine against Salmonella enterica serovar Enteritidis infection.ResultsThe Salmonella strain C50336 with a deletion of the pipC gene exhibits a significant reduction in its ability to resist environmental stress and virulence. Meanwhile, the expression levels of SPI-1-related genes (invH, sipA, sipB, sipC, sopB, and sopE2) and SPI-2-related genes (spvB, ssrA, orf245, ssaS, ssaT, ssaU, sseB, and sseD) encoding the Salmonella type III secretion system (T3SS) were found to be decreased, leading to a significant reduction in the bacteria’s invasion and intracellular survival abilities. The results of the mouse intraperitoneal challenge experiment showed that compared with the wild-type strain, the 50% lethal dose (LD50) of the ΔpipC strain increased by 47 times, and the bacterial loads in the liver, spleen, and cecum were significantly reduced. When mice were immunized with the ΔpipC mutant strain, the immunized mice showed a robust immune response, with significantly increased cytokine and antibody levels in their bodies. Mice vaccinated with the ΔpipC mutant strain had 100% immune protection against wild-type Salmonella infection.ConclusionThis study demonstrates that lack of pipC affects SE pathogenicity by decreasing its virulence both in vitro and in vivo. Vaccination of mice with ΔpipC conferred development of an acquired immunity and efficacious protection against experimental systemic infection. These results indicated that the ΔpipC mutant strain can be used in the development of attenuated live vaccines.
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spelling doaj-art-7a1b8b6052a54083a9e72e6d92e2cae62025-08-20T03:16:16ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-06-011610.3389/fmicb.2025.16310081631008PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in miceLu Zhang0Yubin Chen1Zhigang Yan2Yuntai Li3Xiaowen Yang4Li Chen5Yanying Zhang6Yingyu Chen7Yonghui Li8Qiumei Shi9Tonglei Wu10Hebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaHebei Provincial Center for Livestock Breeding Improvement, Shijiazhuang, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaKey Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaCollege of Animal Medicine, Huazhong Agricultural University, Wuhan, ChinaThe Second Hospital of Qinhuangdao, Qinhuangdao, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaHebei Provincial Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao, ChinaBackgroundSalmonellosis caused by Salmonella sp. is a foodborne zoonotic disease that poses a significant threat to public health security. Vaccination is a safe and effective strategy for preventing and controlling Salmonella infections. PipC is a chaperone protein associated with Salmonella invasion proteins which is crucial for bacteria to invade host cells.MethodsIn this study, a ΔpipC mutant strain was generated. Subsequently, we examined the environmental stress tolerance of the mutant strain through in vitro simulation experiments. Moreover, its virulence by employing cell and mouse infection models was investigated. Furthermore, we utilized a mouse model to further explore its potential as an attenuated live vaccine against Salmonella enterica serovar Enteritidis infection.ResultsThe Salmonella strain C50336 with a deletion of the pipC gene exhibits a significant reduction in its ability to resist environmental stress and virulence. Meanwhile, the expression levels of SPI-1-related genes (invH, sipA, sipB, sipC, sopB, and sopE2) and SPI-2-related genes (spvB, ssrA, orf245, ssaS, ssaT, ssaU, sseB, and sseD) encoding the Salmonella type III secretion system (T3SS) were found to be decreased, leading to a significant reduction in the bacteria’s invasion and intracellular survival abilities. The results of the mouse intraperitoneal challenge experiment showed that compared with the wild-type strain, the 50% lethal dose (LD50) of the ΔpipC strain increased by 47 times, and the bacterial loads in the liver, spleen, and cecum were significantly reduced. When mice were immunized with the ΔpipC mutant strain, the immunized mice showed a robust immune response, with significantly increased cytokine and antibody levels in their bodies. Mice vaccinated with the ΔpipC mutant strain had 100% immune protection against wild-type Salmonella infection.ConclusionThis study demonstrates that lack of pipC affects SE pathogenicity by decreasing its virulence both in vitro and in vivo. Vaccination of mice with ΔpipC conferred development of an acquired immunity and efficacious protection against experimental systemic infection. These results indicated that the ΔpipC mutant strain can be used in the development of attenuated live vaccines.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1631008/fullSalmonella enterica serovar EnteritidisPipCvirulenceimmune protectionvaccine
spellingShingle Lu Zhang
Yubin Chen
Zhigang Yan
Yuntai Li
Xiaowen Yang
Li Chen
Yanying Zhang
Yingyu Chen
Yonghui Li
Qiumei Shi
Tonglei Wu
PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
Frontiers in Microbiology
Salmonella enterica serovar Enteritidis
PipC
virulence
immune protection
vaccine
title PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
title_full PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
title_fullStr PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
title_full_unstemmed PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
title_short PipC affects the virulence of Salmonella enterica serovar Enteritidis and its deletion strain provides effective immune protection in mice
title_sort pipc affects the virulence of salmonella enterica serovar enteritidis and its deletion strain provides effective immune protection in mice
topic Salmonella enterica serovar Enteritidis
PipC
virulence
immune protection
vaccine
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1631008/full
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