Immunization with a combination of recombinant <i>Brucella abortus</i> proteins induces T helper immune response and confers protection against wild‐type challenge in BALB/c mice

Li, Zhiqiang; Wang, Shuli; Wei, Shujuan; Yang, Guangli; Zhang, Chunmei; Li, Xi; Zhang, Jinliang; Cui, Yanyan; Hao, Junfang; Zhang, Huan; Zhang, Hui

doi:10.1111/1751-7915.14015

Cited by 6 publications

(5 citation statements)

References 58 publications

(74 reference statements)

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“…Indeed, the acquisition of Brucella infection occurs due to mucosal exposure. Currently, immunization is a very successful strategy for preventing and managing animal brucellosis (Li et al 2022 ). When administered to animals, the optimal vaccine for brucellosis should possess avirulent or attenuated characteristics, effectively preventing Brucella infections.…”

Section: Discussionmentioning

confidence: 99%

“…Simultaneously, it should not impede the accuracy of serological diagnostic methods (Heidary et al 2022 ). Furthermore, the intervention must have the capability to effectively inhibit abortion and virulence reversal while concurrently facilitating extended durations of immune protection (Li et al 2022 ). Nevertheless, contemporary vaccinations have significant drawbacks that restrict their use, notwithstanding their efficacy in averting animal infections.…”

Section: Discussionmentioning

confidence: 99%

“…Hence, the endeavor to create a vaccination that is both safe and productive is a challenging undertaking. Outer membrane proteins (OMPs) are a subset of surface antigens found in Brucella (Li et al 2022 ; Heidary et al 2022 ). One may argue that oligomeric membrane proteins (OMPs) possess favorable attributes that provide viable options for producing recombinant proteins intended for both preventive and diagnostic purposes (Heidary et al 2022 ).…”

Section: Discussionmentioning

confidence: 99%

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Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Kazemi-Roudsari,

Doosti,

Jami

2024

AMB Expr

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Brucellosis is regarded as one of the world’s most severe zoonotic diseases. This study aimed to investigate the possibility of using recombinant Lactococcus lactis (L. lactis) as a live vector to produce recombinant Brucella abortus (B. abortus) Omp10. The gene sequences were obtained from GenBank. The proteins’ immunogenicity was assessed using Vaxijen. After confirming the cloning of the Omp10 gene in the pNZ8148 vector by enzymatic digestion and PCR, transformation into L. lactis was done. SDS-PAGE and western blot methods evaluated omp10 protein expression. Mice received oral recombinant L. lactis vaccines. IgG antibodies against Omp10 were tested using ELISA. Real-time PCR and ELISA were used to analyze cytokine responses. Survival rate and histopathological changes were evaluated after the challenge. Omp10 was chosen for its 1.5524 antigenicity score. Enzymatic digestion and PCR identified a 381-bp gene fragment. A 10 kDa band indicated the success of L. lactis transformation. Mice administered the L. lactis-pNZ8148-Omp10-Usp45 vaccination 14 days after priming showed significantly higher Omp10-specific total IgG and IgG1 (P < 0.001) than the PBS control group. The mice who received the L. lactis-pNZ8148-Omp10-Usp45 and IRBA vaccines had significantly elevated levels of IFN-γ, TNFα, IL-4, and IL-10 in samples collected on days 14 and 28 (P < 0.001). Inflammatory response, morphological damage, alveolar edema, and lymphocyte infiltration were reduced in the target group. A recombinant L. lactis expressing the Omp10 protein was constructed as an oral Lactococcus-based vaccine and compared to live attenuated vaccines for future brucellosis investigations.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Kazemi-Roudsari,

Doosti,

Jami

2024

AMB Expr

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“…The discovery of immunogenic proteins that may elicit a cell‐mediated immune response, which is necessary to fight the pathogen's intracellular location, has been the foundation of current techniques for developing novel vaccines vs. B. abortus (Piri‐Gharaghie, Beiranvand, et al., 2022 ; Piri‐Gharaghie, Doosti, et al., 2022 ). The omp25, the antigen of B. abortus , can boost the immune defence (Li et al., 2022 ). Mice that received immunization with pure omp25 showed exceptional protection against B. abortus infection (Zhang et al., 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Brucella abortus antigen omp25 vaccines: Development and targeting based on Lactococcus lactis

Gouran,

Doosti,

Jami

2023

Veterinary Medicine & Sci

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Background Most Brucella infections take place on mucosal membranes. Therefore, creating vaccinations delivered through the mucosa may be crucial for managing brucellosis. Consequently, we assessed the efficacy of a recombinant oral antigen delivery system based on Lactococcus lactis for Brucella abortus omp25 antigen. Method Oral vaccinations with L. lactis transformed with pNZ8148 variants encoding for omp25 (pNZ8148:omp25) and free‐pNZ8148 were administered to mice. On day 30, following immunization in animal groups, anti‐omp25‐specific IgG1 antibodies were assessed by the ELISA test. Additionally, nasal and bronchoalveolar lavages containing omp25‐specific secretory IgA (sIgA) were analysed by ELISA. ELISA test and real‐time PCR were also used to analyse cytokine responses up to 28 days following the last boost. In addition, the protective potential of L. lactis pNZ8148:omp25 vaccines was assessed in BALB/c mice by exposing them to the B. abortus strain. Results Based on the initial screening results, the omp25 protein was identified for immunogenicity because it had the maximum solubility and flexibility and antigenic values of 0.75. The produced plasmid was digested using KpnI and XbaI. By electrophoretic isolation of the digestion fragments at 786 bp, the omp25 gene, the successful production of the recombinant plasmid, was confirmed. Antigen expression at the protein level revealed that the target group generated the 25 kDa‐sized omp25 protein, but there was no protein expression in the control group. Fourteen days after priming, there was a considerable amount of omp25‐specific IgG1 in the sera of mice vaccinated with pNZ8148–Usp45–omp25–L. lactis (p < 0.001 in target groups compared to the phosphate‐buffered saline control group). IFN‐γ and TNF‐α levels were more significant in samples from mice that had been given the pNZ8148–Usp45–omp25–L. lactis and IRBA vaccinations, in samples taken on days 14 and 28, respectively (p < 0.001). The pNZ8148–Usp45–omp25–L. lactis and IRBA immunization groups had significantly greater IL‐4 and IL‐10 transcription levels than the other groups. The spleen portions from the pNZ8148–Usp45–omp25–L. lactis and IRIBA vac group had less extensive spleen injuries, alveolar oedema, lymphocyte infiltration and morphological damage due to the inflammatory process. Conclusion Our study offers a novel method for using the food‐grade, non‐pathogenic and noncommercial bacterium L. lactis as a protein cell factory to produce the novel immunogenic fusion candidate romp25. This method offers an appealing new approach to assessing the cost‐effective, safe, sustainable, simple pilot development of pharmaceutical products.

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“…Recently the advances in genetic engineering to substitute S-LPS resulted in the development of the novel ELISA tests based on the application of non-LPS Brucella immunodominant antigens either as a pure natural protein or multi-epitope recombinant proteins [ 8 , 10 – 12 ]. Among them, Brucella outer membrane proteins with different molecular weight (groups I, II, and III with MW of 88–94, 36–38, and 25–34 kDa, respectively) have been recognized as potential candidates for application in brucellosis diagnosis techniques or vaccine design [ 8 , 10 , 12 , 13 ]. For example, outer membrane protein 19 kDa (OMP19), which is conserved among most Brucella species, have been proven to be capable of conferring immunity in mice [ 12 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

Serodiagnosis of human brucellosis by an indirect ELISA test using recombinant outer membrane protein 19 kDa (rOMP19) as an antigen

Golchin,

Mollayi,

Mohammadi

et al. 2023

BMC Biotechnol

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Background Brucellosis remains one of the global health concerns that reemerges in recent years. Delayed or inaccurate diagnosis end to a long treatment duration and financial burden; therefore, finding a good antigen for detection of specific anti-Brucella antibodies is crucial. We intended to evaluate the serodiagnosis value of recombinant Brucella outer membrane protein 19 kDa (rOMP19) using indirect ELISA system compared with Rose Bengal test. Results The OMP19 sequence was successfully cloned into pET-28a and produced in E. coli cells (DE3). After extraction and purification of rOMP19, this protein was used for designing indirect ELISA to detect anti-Brucella antibodies in 73 human sera, including 6 brucellosis-positive and 67 brucellosis-negative samples. The accuracy of rOMP19 ELISA was evaluated by receiver operating characteristic (ROC) curve and then compared with Rose Bengal plate test and a commercial anti-IgG Brucella ELISA kit. In comparison with Rose Bengal plate test, the area under the ROC curve was 0.985 (95% CI, 0.96–1.00). From coordinates of the curve, the optimal cut-off value was selected at 0.147, in which the diagnostic sensitivity was 100%, and the specificity was 94%. At this cut-off point, 10 samples were diagnosed as positive (6 true positives and 4 false positives), while negative samples were all correctly diagnosed. The results of our designed rOMP19 ELISA was the same as data obtained from commercial ELISA kit, which applied LPS as an antigen. Conclusions We concluded that OMP19 is an efficient antigen for the serodiagnosis of human brucellosis.

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Immunization with a combination of recombinant Brucella abortus proteins induces T helper immune response and confers protection against wild‐type challenge in BALB/c mice

Cited by 6 publications

References 58 publications

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Brucella abortus antigen omp25 vaccines: Development and targeting based on Lactococcus lactis

Serodiagnosis of human brucellosis by an indirect ELISA test using recombinant outer membrane protein 19 kDa (rOMP19) as an antigen

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