Screening of Mycoplasma hyopneumoniae Subunit Vaccine Candidate Antigens Using Multiple Immunological Approaches

Description

This study tested the hypothesis that Mycoplasma hyopneumoniae proteins predicted to have favorable antigenic features may include immunogenic candidates for subunit vaccine development. Recombinant Mhp proteins were selected by bioinformatic analysis, expressed and purified, and screened by immunoblotting with sera from Mhp-infected pigs; selected proteins were further evaluated using IFN-γ ELISpot assays with porcine PBMCs and mouse immunization experiments assessing humoral, mucosal, and cellular immune responses. Six proteins, Mhp P2, Mhp VIR1, Mhp P10, Mhp P17, Mhp MEM1, and Mhp RED1, showed measurable immunogenicity across the screening process, with Mhp P17 inducing the strongest humoral and mucosal responses in mice. These data support the prioritization of these proteins as candidate antigens for further validation, but they should be interpreted as evidence of immunogenic potential rather than confirmed protective efficacy in pigs.

Steps to reproduce

The data were generated through an integrated antigen-screening workflow for Mycoplasma hyopneumoniae subunit vaccine candidate identification. Candidate Mhp proteins were first selected using bioinformatic prediction tools, including LipoP 1.0, SignalP 6.0, TMHMM 2.0, and VaxiJen 2.0, to evaluate antigen-related features such as lipoprotein signals, signal peptides, transmembrane regions, and predicted antigenicity. Selected genes were cloned, expressed in an Escherichia coli recombinant expression system, and the resulting proteins were purified by affinity chromatography. The purified recombinant proteins were then screened by immunoblotting using sera from Mhp-infected pigs to identify antigens recognized during infection. Proteins showing immunoreactivity were further evaluated for their ability to stimulate cellular immune responses using IFN-γ ELISpot assays with porcine peripheral blood mononuclear cells. Finally, selected immunogenic proteins were used to immunize mice, and antigen-specific humoral, mucosal, and cellular immune responses were assessed by standard immunological assays. This workflow allows the dataset to be used for comparing the relative immunogenicity of recombinant Mhp proteins and for prioritizing candidate antigens for subsequent protective-efficacy studies in pigs.

Institutions

• Huazhong Agricultural University

  Hubei, Wuhan

Categories

Licence