title: Sonicated Protein Fractions of Mycoplasma hyopneumoniae Induce Inflammatory Responses and Differential Gene Expression in a Murine Alveolar Macrophage Cell Line.
abstract: Mycoplasma hyopneumoniae is known to cause porcine enzootic pneumonia (EP), an important disease in swine production. The objective of this study was to examine the effects of sonicated protein fractions of M. hyopneumoniae on inflammatory response and gene expression in the murine alveolar macrophage MH-S cell line. The effects of sonicated protein fractions and intact M. hyopneumoniae on the gene expression of cytokines and iNOS were assessed using RT-PCR. The Annealing Control Primer (ACP)-based PCR method was used to screen differentially expressed genes. Increased transcription of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, COX-2, and iNOS mRNA was observed after exposure to the supernatant (SPT), precipitant (PPT), and intact M. hyopneumoniae protein. A time-dependent analysis of the mRNA expression revealed an upregulation after 4 h for IL-6 and iNOS and after 12 h for IL-1β and TNF-α, for both SPT and PPT; the fold change in COX-2 expression was less. A dose- and time-dependent correlation was observed in nitrite (NO) production for both protein fractions; however, there was no significant difference between the effects of the two protein fractions. In a differential gene analysis, PCR revealed differential expression for nine gene bands after 3 h of stimulation - only one gene was downregulated, while the remaining eight were upregulated. The results of this study provide insights that help improve our understanding of the mechanisms underlying the pathogenesis of and macrophage defenses against M. hyopneumoniae assault, and suggest targets for future studies on therapeutic interventions for M. hyopneumoniae infections.
authors.affiliation: Laboratory of Veterinary Pharmacokinetics and Pharmacodynamics, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.|Department of Biological Chemistry, The Weizmann Institute of Science, 76100 Rehovot, Israel.
authors.forename: Dereje
authors.initials: D
authors.lastname: Damte
authors.affiliation: Laboratory of Veterinary Pharmacokinetics and Pharmacodynamics, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Seung-Jin
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Laboratory of Veterinary Pharmacokinetics and Pharmacodynamics, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Biruk Tesfaye
authors.initials: BT
authors.lastname: Birhanu
authors.affiliation: Center for Nutraceutical and Pharmaceutical Materials, Division of Bioscience and Bioinformatics, Science Campus, Myongji University, Yongin 449-728, Republic of Korea.
authors.forename: Joo-Won
authors.initials: JW
authors.lastname: Suh
authors.affiliation: Laboratory of Veterinary Pharmacokinetics and Pharmacodynamics, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Seung-Chun
authors.initials: SC
authors.lastname: Park
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology* / Q000302:isolation & purification; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D007249:Inflammation / Q000139:chemically induced*; D016676:Macrophages, Alveolar / Q000187:drug effects*; D051379:Mice; D045705:Mycoplasma hyopneumoniae / Q000737:chemistry*; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D016207:Cytokines; D012333:RNA, Messenger; D052247:Nitric Oxide Synthase Type II
keywords: Cytokine; M. hyopneumoniae; nitric oxide; sonicated protein fraction