Infection-induced antibodies against the major outer membrane protein of Campylobacter jejuni mainly recognize
Shouxiong Huang1, Orhan Sahin, Qijing Zhang
1Food Animal Health Research Program, Ohio Agriculture Research and Development Center, The Ohio State University,Wooster, OH, USA.
Abstract:
The major outer membrane protein (MOMP) of Campylobacter jejuni is an abundant surface protein with a pore-forming function and may be a potential candidate for vaccine development. Despite the fact that MOMP is immunogenic and the recombinant MOMP (rMOMP) can be readily produced in Escherichia coli, the nature of the antibody response to MOMP during in vivo infection is not well understood. In this study, various methods involving detergent replacement and liposome reconstitution were used to refold rMOMP, and antibody responses to MOMP elicited in Campylobacter-colonized chickens were evaluated using sera from chickens either naturally or experimentally infected by C. jejuni. The results demonstrated that proteoliposomes restored the reactivity of rMOMP to rabbit antibodies elicited by native MOMP, indicating the recovery of native MOMP conformation by this refolding method. Importantly, sera from naturally or experimentally infected chickens reacted weakly with denatured rMOMP, but strongly with rMOMP reconstituted in proteoliposome, suggesting that the chicken antibody response to MOMP is predominantly directed against conformational epitopes. These observations provide direct evidence for conformation-dependent humoral responses to MOMP induced by Campylobacter infection, demonstrate that C. jejuni MOMP is immunogenic in its natural host and suggest that proteoliposomes may be potentially used for the evaluation of rMOMP-based vaccines.
Insights
Antibody responses to Campylobacter jejuni's major outer membrane protein (MOMP) are conformation-dependent. Refolding recombinant MOMP (rMOMP) in proteoliposomes better mimics native protein structure for vaccine evaluation.
Area of Science:
- Microbiology
- Immunology
- Vaccine Development
Background:
- Campylobacter jejuni major outer membrane protein (MOMP) is abundant, pore-forming, and a potential vaccine candidate.
- The immunogenicity of MOMP and production of recombinant MOMP (rMOMP) are established, but in vivo antibody responses are poorly understood.
Purpose of the Study:
- To investigate the nature of antibody responses to Campylobacter jejuni MOMP during natural and experimental infections in chickens.
- To evaluate refolding methods for rMOMP and their impact on antibody recognition.
Main Methods:
- Refolding of recombinant MOMP (rMOMP) using detergent replacement and liposome reconstitution.
- Evaluation of antibody responses in chicken sera against native and refolded rMOMP.
Main Results:
- Proteoliposome reconstitution restored the reactivity of rMOMP to antibodies against native MOMP.
- Chicken sera showed weak reactivity to denatured rMOMP but strong reactivity to proteoliposome-reconstituted rMOMP.
- Antibody responses in chickens are primarily directed against conformational epitopes of MOMP.
Conclusions:
- Humoral immune responses to C. jejuni MOMP are conformation-dependent in the natural host.
- C. jejuni MOMP is immunogenic in chickens, eliciting conformation-specific antibodies.
- Proteoliposomes show potential for evaluating rMOMP-based vaccines.
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