Identification of Novel Seroreactive Antigens in Johne’s Disease Cattle by Using the Mycobacterium tuberculosis Protein Array

Authors

John Bannantine, USDA ARS National Animal Disease CenterFollow
Joseph J. Campo, Antigen Discovery, Inc., Irvine
Lingling Li, Pennsylvania State UniversityFollow
Arlo Randall, Antigen Discovery, Inc., Irvine
Jozelyn Pablo, Antigen Discovery, Inc., Irvine
Craig A. Praul, Pennsylvania State University, University Park
Juan Antonio Raygoza Garay, Pennsylvania State University, University Park
Judith R. Stabel, USDA-ARS, National Animal Disease CenterFollow
Vivek Kapur, Pennsylvania State UniversityFollow

ORCID IDs

http://orcid.org/0000-0002-5692-7898

Document Type

Article

Date of this Version

2017

Citation

Bannantine JP, Campo JJ, Li L, Randall A, Pablo J, Praul CA, Raygoza Garay JA, Stabel JR, Kapur V. 2017. Identification of novel seroreactive antigens in Johne's disease cattle by using the Mycobacterium tuberculosis protein array. Clin Vaccine Immunol 24:e00081-17. https://doi.org/10 .1128/CVI.00081-17.

Comments

U.S. government works are not subject to copyright.

Abstract

Johne’s disease, a chronic gastrointestinal inflammatory disease caused by Mycobacterium avium subspecies paratuberculosis, is endemic in dairy cattle and other ruminants worldwide and remains a challenge to diagnose using traditional serological methods. Given the close phylogenetic relationship between M. aviumsubsp. paratuberculosis and the human pathogen Mycobacterium tuberculosis, here, we applied a whole-proteome M. tuberculosis protein array to identify seroreactive and diagnostic M. avium subsp. paratuberculosis antigens. A genome-scale pairwise analysis of amino acid identity levels between orthologous proteins in M. avium subsp. paratuberculosis and M. tuberculosis showed an average of 62% identity, with more than half the orthologous proteins sharing 􏰀75% identity. Analysis of the M. tuberculosis protein array probed with sera from M. avium subsp. paratuberculosis- infected cattle showed antibody binding to 729 M. tuberculosis proteins, with 58% of them having 􏰄70% identity to M. avium subsp. paratuberculosis orthologs. The results showed that only 4 of the top 40 seroreactive M. tuberculosis antigens were orthologs of previously reported M. avium subsp. paratuberculosis antigens, revealing the existence of a large number of previously unrecognized candidate diagnostic antigens. Enzyme-linked immunosorbent assay (ELISA) testing of 20 M. avium subsp. paratuberculosis recombinant proteins, representing reactive and nonreactive M. tuberculosis orthologs, further confirmed that the M. tuberculosis array has utility as a screening tool for identifying candidate antigens for Johne’s disease diagnostics. Additional ELISA testing of field serum samples collected from dairy herds around the United States revealed that MAP2942c had the strongest seroreactivity with Johne’s disease-positive samples. Collectively, our studies have considerably expanded the number of candidate M. avium subsp. paratuberculosis proteins with potential utility in the next generation of rationally designed Johne’s disease diagnostic assays.