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Protein-specific analysis of humoral immune responses in a clinical trial for vaccines against contagious bovine pleuropneumonia.

ABSTRACT: Specific humoral immune responses in a clinical trial on cattle for vaccines against contagious bovine pleuropneumonia (CBPP) were investigated. The trial included a subunit vaccine consisting of five recombinant putative variable surface proteins of the infectious agent Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) compared to the currently approved attenuated vaccine strain T1/44 and untreated controls. Humoral immune responses to 65 individual recombinant surface proteins of M. mycoides SC were monitored by a recently developed bead-based array assay. Responses to the subunit vaccine components were found to be weak. Animals vaccinated with this vaccine were not protected and had CBPP lesions similar to those of the untreated controls. In correlating protein-specific humoral responses to T1/44-induced immunity, five proteins associated with a protective immune response were identified by statistical evaluation, namely, MSC_1046 (LppQ), MSC_0271, MSC_0136, MSC_0079, and MSC_0431. These five proteins may be important candidates in the development of a novel subunit vaccine against CBPP.

SUBMITTER: Hamsten C

PROVIDER: S-EPMC2863394 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature

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Protein-specific analysis of humoral immune responses in a clinical trial for vaccines against contagious bovine pleuropneumonia.

Hamsten Carl C Tjipura-Zaire Georgina G McAuliffe Laura L Huebschle Otto J B OJ Scacchia Massimo M Ayling Roger D RD Persson Anja A

Clinical and vaccine immunology : CVI 20100331 5

Specific humoral immune responses in a clinical trial on cattle for vaccines against contagious bovine pleuropneumonia (CBPP) were investigated. The trial included a subunit vaccine consisting of five recombinant putative variable surface proteins of the infectious agent Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) compared to the currently approved attenuated vaccine strain T1/44 and untreated controls. Humoral immune responses to 65 individual recombinant surface prot ...[more]

PMID: 20357055

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Project description:AimsThe aim of this study was to analyse the current situation of Contagious Bovine Pleuropneumonia (CBPP) in the Central African Republic (CAR) by seroprevalence analysis, as well as isolation and characterization of strains of the etiologic agent, Mycoplasma mycoides mycoides (Mmm), circulating in livestock breeding regions.Material and methodsThe strains obtained were subjected to whole genome sequencing by Illumina technology and genotyped using the eMLST technique based on 62 genes of the Mmm core genome. Their sensitivity to tetracycline was assessed by determination of the minimum inhibitory concentration (MIC) on agar. A seroprevalence analysis by competitive ELISA was conducted in livestock breeding regions (West, Centre and East CAR), including both zebu and taurine cattle breeds, and both males and females.ResultsThe three strains isolated in the three regions of the CAR shared exactly the same genomic sequence. Phylogenetic analysis showed that they were closely related to a strain isolated in the CAR in 1991, also sequenced in this study, and clustered with Mmm strains originating from East and Central Africa. The recent isolates presented increased MIC values, though they were still sensitive to tetracycline. The global CBPP prevalence in the CAR was estimated at 12.5% with no significant differences observed between cattle breeding regions, nor between males and females. However, a significantly higher prevalence was observed in zebu compared to taurine cattle, most likely in relation to their herding system based on cattle transhumance and nomadic pastoralism.ConclusionCBPP is enzootic in the CAR in spite of control campaigns based on use of the live T1 vaccines, which have shown little efficacy due to poor implementation in the field. New strategies combining controlled use of antibiotics and inactivated vaccines, with increased thermostability, should be well received by livestock keepers and allow a better control of CBPP in the region. The fact that the recent Mmm isolates are still resistant to tetracycles is encouraging.

Project description:For the development of a competitive ELISA (cELISA) to detect serum antibodies against the Mycoplasma mycoides subsp. Mycoides (Mmm) (strain PG1), the causative agent of contagious bovine pleuropneumonia (CBPP), all the proteins of this pathogen were analyzed. Then, a specific extracellular region of a transmembrane protein with the potential for diagnosis was identified. After that, a monoclonal antibody (Mab) named 3A8 was obtained using this extracellular region as an immunogen. Finally, a cELISA was established with the extracellular domain of this transmembrane protein as the coating antigen, Mab 3A8 as the competitive antibody, and HRP-labeled goat anti-mouse IgG as the enzyme-labeled antibody. This established method was used to detect the antibody dynamic regularity of goats which are artificially immunized Mmm and was also compared with a commercial ELISA kit. Further, the sera of 1011 different cattle from border provinces of China were monitored using a candidate Mab 3A8 cELISA. The detection results of known background sera used in this study indicate that a candidate diagnostic marker was successfully identified by analyzing all the coding proteins of Mmm in this research, and the cELISA established based on the Mab 3A8 against this protein can detect CBPP-positive serum with specificity and has no cross-reaction with other related epidemic disease-positive sera. In addition, we tested the sera collected from the border areas of China using the established ELISA, and no positive sample was detected. The research protocol of the CBPP cELISA established in this study is different from the traditional method, which can greatly reduce the investment of manpower and capital and save development time. We believe that this study's protocol could serve as a reference for the development of detection methods for mycoplasma and other complex pathogens. KEY POINTS: • A Mmm-specific diagnostic marker was obtained based on protein characteristics. • A cELISA was established for CBPP serum antibody detection. • The serological investigation was conducted for CBPP in the border areas of China.

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Protein-specific analysis of humoral immune responses in a clinical trial for vaccines against contagious bovine pleuropneumonia.

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Protein-specific analysis of humoral immune responses in a clinical trial for vaccines against contagious bovine pleuropneumonia.

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