title: Elongation Factor Tu and Heat Shock Protein 70 Are Membrane-Associated Proteins from Mycoplasma ovipneumoniae Capable of Inducing Strong Immune Response in Mice.
abstract: Chronic non-progressive pneumonia, a disease that has become a worldwide epidemic has caused considerable loss to sheep industry. Mycoplasma ovipneumoniae (M. ovipneumoniae) is the causative agent of interstitial pneumonia in sheep, goat and bighorn. We here have identified by immunogold and immunoblotting that elongation factor Tu (EF-Tu) and heat shock protein 70 (HSP 70) are membrane-associated proteins on M. ovipneumonaiea. We have evaluated the humoral and cellular immune responses in vivo by immunizing BALB/c mice with both purified recombinant proteins rEF-Tu and rHSP70. The sera of both rEF-Tu and rHSP70 treated BALB/c mice demonstrated increased levels of IgG, IFN-γ, TNF-α, IL-12(p70), IL-4, IL-5 and IL-6. In addition, ELISPOT assay showed significant increase in IFN-γ+ secreting lymphocytes in the rHSP70 group when compared to other groups. Collectively our study reveals that rHSP70 induces a significantly better cellular immune response in mice, and may act as a Th1 cytokine-like adjuvant in immune response induction. Finally, growth inhibition test (GIT) of M. ovipneumoniae strain Y98 showed that sera from rHSP70 or rEF-Tu-immunized mice inhibited in vitro growth of M. ovipneumoniae. Our data strongly suggest that EF-Tu and HSP70 of M. ovipneumoniae are membrane-associated proteins capable of inducing antibody production, and cytokine secretion. Therefore, these two proteins may be potential candidates for vaccine development against M. ovipneumoniae infection in sheep.
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Fei
authors.initials: F
authors.lastname: Jiang
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Jinyan
authors.initials: J
authors.lastname: He
authors.affiliation: Center For Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, United States of America.
authors.forename: Nalu
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xu
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.|Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Wenxue
authors.initials: W
authors.lastname: Wu
mesh_terms: D000818:Animals; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D007111:Immunity, Cellular / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000097:blood; D018664:Interleukin-12 / Q000097:blood; D015847:Interleukin-4 / Q000097:blood; D015848:Interleukin-5 / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D045802:Mycoplasma ovipneumoniae / Q000276:immunology* / Q000378:metabolism; D010444:Peptide Elongation Factor Tu / Q000276:immunology*; D011019:Pneumonia, Mycoplasma / Q000276:immunology / Q000662:veterinary*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D018840:HSP70 Heat-Shock Proteins; D007074:Immunoglobulin G; D015848:Interleukin-5; D015850:Interleukin-6; D008565:Membrane Proteins; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D015847:Interleukin-4; D007371:Interferon-gamma; D010444:Peptide Elongation Factor Tu