title: Mycobacterium avium Subspecies paratuberculosis Recombinant Proteins Modulate Antimycobacterial Functions of Bovine Macrophages.
abstract: It has been shown that Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) activates the Mitogen Activated Protein Kinase (MAPK) p38 pathway, yet it is unclear which components of M. paratuberculosis are involved in the process. Therefore, a set of 42 M. paratuberculosis recombinant proteins expressed from coding sequences annotated as lipoproteins were screened for their ability to induce IL-10 expression, an indicator of MAPKp38 activation, in bovine monocyte-derived macrophages. A recombinant lipoprotein, designated as MAP3837c, was among a group of 6 proteins that strongly induced IL-10 gene transcription in bovine macrophages, averaging a 3.1-fold increase compared to non-stimulated macrophages. However, a parallel increase in expression of IL-12 and TNF-α was only observed in macrophages exposed to a subset of these 6 proteins. Selected recombinant proteins were further analyzed for their ability to enhance survival of M. avium within bovine macrophages as measured by recovered viable bacteria and nitrite production. All 6 IL-10 inducing MAP recombinant proteins along with M. paratuberculosis cells significantly enhanced phosphorylation of MAPK-p38 in bovine macrophages. Although these proteins are likely not post translationally lipidated in E. coli and thus is a limitation in this study, these results form the foundation of how the protein component of the lipoprotein interacts with the immune system. Collectively, these data reveal M. paratuberculosis proteins that might play a role in MAPK-p38 pathway activation and hence in survival of this organism within bovine macrophages.
authors.affiliation: National Animal Disease Center, USDA-Agricultural Research Service, Ames, Iowa, United States of America.
authors.forename: John P
authors.initials: JP
authors.lastname: Bannantine
authors.affiliation: National Animal Disease Center, USDA-Agricultural Research Service, Ames, Iowa, United States of America.
authors.forename: Judith R
authors.initials: JR
authors.lastname: Stabel
authors.affiliation: Department of Veterinary Clinical Sciences, Washington State University, Pullman, Washington, United States of America.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Laws
authors.affiliation: Department of Veterinary Clinical Sciences, Washington State University, Pullman, Washington, United States of America.
authors.forename: Maria Clara
authors.initials: MC
authors.lastname: D Cardieri
authors.affiliation: Department of Veterinary Clinical Sciences, Washington State University, Pullman, Washington, United States of America.
authors.forename: Cleverson D
authors.initials: CD
authors.lastname: Souza
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology*; D002417:Cattle; D016207:Cytokines / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007113:Immunity, Innate / Q000187:drug effects*; D056747:Immunomodulation / Q000187:drug effects*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology* / Q000382:microbiology; D050296:Microbial Viability / Q000187:drug effects / Q000276:immunology; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis; D010587:Phagocytosis; D010766:Phosphorylation; D011994:Recombinant Proteins; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D016207:Cytokines; D011994:Recombinant Proteins; D009569:Nitric Oxide; D048051:p38 Mitogen-Activated Protein Kinases