Identification of conserved Mycoplasma agalactiae surface antigens by immunoproteomics.

Autor: Cacciotto C; Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Italy; Mediterranean Center for Disease Control (MCDC), Università degli studi di Sassari, Italy; Porto Conte Ricerche Srl, Alghero, Italy., Addis MF; Mediterranean Center for Disease Control (MCDC), Università degli studi di Sassari, Italy; Porto Conte Ricerche Srl, Alghero, Italy; Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Italy. Electronic address: filippa.addis@unimi.it., Pagnozzi D; Mediterranean Center for Disease Control (MCDC), Università degli studi di Sassari, Italy., Coradduzza E; Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Italy; Istituto Zooprofilattico Sperimentale della Sardegna 'G. Pegreffi', Sassari, Italy., Pittau M; Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Italy; Mediterranean Center for Disease Control (MCDC), Università degli studi di Sassari, Italy., Alberti A; Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Italy; Mediterranean Center for Disease Control (MCDC), Università degli studi di Sassari, Italy. Electronic address: alberti@uniss.it.
Jazyk: angličtina
Zdroj: Veterinary immunology and immunopathology [Vet Immunol Immunopathol] 2021 Jun; Vol. 236, pp. 110239. Date of Electronic Publication: 2021 Apr 07.
DOI: 10.1016/j.vetimm.2021.110239
Abstrakt: Contagious agalactia represents one of the most relevant infectious diseases of dairy sheep, with Mycoplasma agalactiae being the primary etiological agent. The early, sensitive, and specific identification of infected animals, as well as the development of efficient prophylactic tools, remain challenging. Here, we present a comprehensive characterization of M. agalactiae antigens focusing on those shared among different isolates. Leveraging on previous proteomic data obtained on individual strains, we adopted a strategy entailing sample pooling to optimize the identification of conserved proteins that induce an immune response. The liposoluble proteins from previously characterized field isolates and the type strain PG2 T were enriched by Triton X-114 fractionation, pooled, analysed by one-dimensional (1D) and two-dimensional (2D) electrophoresis, and subjected to western immunoblotting against sheep sera collected during natural infection with M. agalactiae. Immunodominant antigens were identified by Matrix-Assisted Laser Desorption-Time-Of-Flight-Mass Spectrometry (MALDI-TOF-MS). This combined immunoproteomic approach confirmed the role of several known immunogens, including P80, P48, and P40, and most variable surface proteins (Vpmas), and unveiled novel immunodominant, conserved antigens, including MAG_1000, MAG_2220, MAG_1980, phnD, MAG_4740, and MAG_2430. Genomic context, functional prediction, subcellular localization, and invariable expression of these proteins in all isolates suggest their possible involvement in bacterial pathogenicity and metabolism. Moreover, most of the identified antigens elicit a host humoral response since the early stages of infection, persisting for at least 270 days. The immunodominant, conserved antigen panel identified in this work supports the development of effective vaccines and diagnostic tools with higher sensitivity and specificity in all the natural infection stages.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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