Construction and characterization of an OmpH-deficient mutant of <i>Pasteurella multocida</i> strain X-73

Varinrak, Thanya; Muenthaisong, Anucha; Apinda, Nisachon; Sawada, Takuo; Sthitmatee, Nattawooti

doi:10.1080/03079457.2018.1533239

Cited by 7 publications

(5 citation statements)

References 25 publications

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“…It has been shown that deletion of the OmpH gene of P. multocida is capable of destroying the bacterial capsule. The deletion of the OmpH gene resulted in a significant reduction in toxicity and the deletion strain provided a good protection against both chickens and mice [ 9 ]. This is consistent with our study—by injecting WTP0910 and ΔompH in Qinghai yaks, we found that the Qinghai yaks injected with ΔompH at the minimum lethal dose of P0910 were able to survive.…”

Section: Discussionmentioning

confidence: 99%

“…Bioinformatics analyses were performed as previously described by Jia [ 9 ]. Functional annotation and classification of all the identified proteins were performed using the Blast2GO program, based on the data in the uniport database.…”

Section: Methodsmentioning

confidence: 99%

“…The outer membrane protein H (OmpH) , among the most important outer membrane proteins of P. multocida , plays a key role in the pathogenicity and invasiveness of the bacteria [ 7 ]. The OmpH gene Pasteurella multicida of poultry origin is a 1062 bp fragment and displays 99.9% homology with the cp39 gene that codes for the capsule protein [ 8 , 9 ]. OmpH gene deletion significantly reduces the resistance of the mutant bacteria to phagocytosis.…”

Section: Introductionmentioning

confidence: 99%

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The Immunoprotection of OmpH Gene Deletion Mutation of Pasteurella multocida on Hemorrhagic Sepsis in Qinghai Yak

Jia

Zhao

et al. 2023

Veterinary Sciences

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OmpH is among the most important virulence factors of Pasteurella multocida, which mediates septicemia in yaks (Bos grunniens I) after infection with the bacteria. In the present study, yaks were infected with wild-type (WT) (P0910) and OmpH-deficient (ΔOmpH) P. multocida strains. The mutant strain was generated through the reverse genetic operation system of pathogens and proteomics technology. The live-cell bacterial count and clinical manifestations of P. multocida infection in Qinghai yak tissues (thymus, lung, spleen, lymph node, liver, kidney, and heart) were analyzed. The expression of differential proteins in the yak spleen under different treatments was analyzed using the marker-free method. We found that compared with the mutant strain, the titer of wild-type strains was significantly higher in tissues. Additionally, compared with other organs, the bacteria titer was significantly higher in the spleen. Compared with the WT p0910 strain, the mutant strain generated milder pathological changes in the tissues of yak. Proteomics analysis revealed that 57 of the 773 proteins expressed in P. multocida were significantly differentially expressed between the ΔOmpH and P0910 groups. Of the 57, 14 were over-expressed, whereas 43 were under-expressed. The differentially expressed proteins in the ΔompH group regulated the ABC transporter (ATP-powered translocation of many substrates across membranes) system, the two-component system, RNA degradation, RNA transcription, glycolysis/gluconeogenesis, biosynthesis of ubiquinone and other terpenoid-quinones, oxidative phosphorylation (citrate cycle) as well as fructose and mannose metabolism. The relationship among 54 significantly regulated proteins was analyzed using STRING. We found that WT P0910 and ΔOmpH of P. multocida infection activated the expression of ropE, HSPBP1, FERH, ATP10A, ABCA13, RRP7A, IL-10, IFN-γ, IL-17A, EGFR, and dnaJ. Overall, deletion of the OmpH gene weakened the virulence but maintained the immunogenicity of P. multocida in yak. The findings of this study provide a strong foundation for the pathogenesis of P. multocida and the management of related septicemia in yaks.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Immunoprotection of OmpH Gene Deletion Mutation of Pasteurella multocida on Hemorrhagic Sepsis in Qinghai Yak

Jia

Zhao

et al. 2023

Veterinary Sciences

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“…Available vaccines showed variable results in protecting ducks against fowl cholera in natural outbreaks [ 2 , 3 ]. The outer membrane protein H (OmpH), a major membrane protein of the P. multocida envelop [ 4 ] and a cross protective antigen among avian P. multocida strains [ 5 , 6 , 7 ], incorporated into flow cholera vaccine candidate, provided the strong protective immunity against P. multocida in chickens [ 5 , 6 , 8 , 9 ], ducks [ 10 , 11 ], and cattle [ 12 , 13 ]. Meanwhile, the live attenuated DEV vaccines are widely used for reduction of diseases impact in ducks [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9 Editing of Duck Enteritis Virus Genome for the Construction of a Recombinant Vaccine Vector Expressing ompH Gene of Pasteurella multocida in Two Novel Insertion Sites

et al. 2022

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Duck enteritis virus (DEV) and Pasteurella multocida, the causative agent of duck plague and fowl cholera, are acute contagious diseases and leading causes of morbidity and mortality in duck. The NHEJ-CRISPR/Cas9-mediated gene editing strategy, accompanied with the Cre–Lox system, have been employed in the present study to show that two new sites at UL55-LORF11 and UL44-44.5 loci in the genome of the attenuated Jansen strain of DEV can be used for the stable expression of the outer membrane protein H (ompH) gene of P. multocida that could be used as a bivalent vaccine candidate with the potential of protecting ducks simultaneously against major viral and bacterial pathogens. The two recombinant viruses, DEV-OmpH-V5-UL55-LORF11 and DEV-OmpH-V5-UL44-44.5, with the insertion of ompH-V5 gene at the UL55-LORF11 and UL44-44.5 loci respectively, showed similar growth kinetics and plaque size, compared to the wildtype virus, confirming that the insertion of the foreign gene into these did not have any detrimental effects on DEV. This is the first time the CRISPR/Cas9 system has been applied to insert a highly immunogenic gene from bacteria into the DEV genome rapidly and efficiently. This approach offers an efficient way to introduce other antigens into the DEV genome for multivalent vector.

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“…The major membrane protein of the P. multocida envelope, the outer membrane protein H (OmpH), is a cross-protective antigen highly conserved among avian P. multocida strains. When incorporated as a fowl cholera vaccine candidate, it provided strong protective immunity against P. multocida in chickens [3][4][5][6][7] and ducks [8,9].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Protective Immune Responses of Ducks against Duck Plague and Fowl Cholera by Recombinant Duck Enteritis Virus Vector Expressing Pasteurella multocida OmpH Gene

et al. 2022

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Duck enteritis virus and Pasteurella multocida are major duck pathogens that induce duck plague and fowl cholera, respectively, in ducks and other waterfowl populations, leading to high levels of morbidity and mortality. Immunization with live attenuated DEV vaccine containing P. multocida outer membrane protein H (OmpH) can provide the most effective protection against these two infectious diseases in ducks. We have recently reported the construction of recombinant DEV expressing P. multocida ompH gene using the CRISPR/Cas9 gene editing strategy with the goal of using it as a bivalent vaccine that can simultaneously protect against both infections. Here we describe the findings of our investigation into the systemic immune responses, potency and clinical protection induced by the two recombinant DEV-ompH vaccine constructs, where one copy each of the ompH gene was inserted into the DEV genome at the UL55-LORF11 and UL44-44.5 intergenic regions, respectively. Our study demonstrated that the insertion of the ompH gene exerted no adverse effect on the DEV parental virus. Moreover, ducklings immunized with the rDEV-ompH-UL55 and rDEV-ompH-UL44 vaccines induced promising levels of P. multocida OmpH-specific as well as DEV-specific antibodies and were completely protected from both diseases. Analysis of the humoral and cellular immunity confirmed the immunogenicity of both recombinant vaccines, which provided strong immune responses against DEV and P. multocida. This study not only provides insights into understanding the immune responses of ducks to recombinant DEV-ompH vaccines but also demonstrates the potential for simultaneous prevention of viral and bacterial infections using viral vectors expressing bacterial immunogens.

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Construction and characterization of an OmpH-deficient mutant of Pasteurella multocida strain X-73

Cited by 7 publications

References 25 publications

The Immunoprotection of OmpH Gene Deletion Mutation of Pasteurella multocida on Hemorrhagic Sepsis in Qinghai Yak

The Immunoprotection of OmpH Gene Deletion Mutation of Pasteurella multocida on Hemorrhagic Sepsis in Qinghai Yak

CRISPR/Cas9 Editing of Duck Enteritis Virus Genome for the Construction of a Recombinant Vaccine Vector Expressing ompH Gene of Pasteurella multocida in Two Novel Insertion Sites

Simultaneous Protective Immune Responses of Ducks against Duck Plague and Fowl Cholera by Recombinant Duck Enteritis Virus Vector Expressing Pasteurella multocida OmpH Gene

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