title: Mycoplasma gallisepticum in vivo induced antigens expressed during infection in chickens.
abstract: Until now only a few genes encoding virulence factors have been characterized in the avian pathogen Mycoplasma gallisepticum. In order to identify candidate targets associated with infection we applied an immunoscreening technique-in vivo induced antigen technology (IVIAT)-to detect immunogens of M. gallisepticum strain Rlow expressed preferentially during in vivo infection. We identified 13 in vivo-induced (IVI) proteins that correspond to different functional categories including: previously reported putative virulence factors (GapA, PlpA, Hlp3, VlhA 1.07 and VlhA 4.01), transport (PotE, MGA_0241 and 0654), translation (L2, L23, ValS), chaperone (GroEL) and a protein with unknown function (MGA_0042). To validate the in vivo antigenic reactivity, 10 IVI proteins were tested by Western blot analysis using serum samples collected from chickens experimentally (with strain Rlow) and naturally (outbreaks, N=3) infected with M. gallisepticum. All IVI proteins tested were immunogenic. To corroborate these results, we tested expression of IVI genes in chickens experimentally infected with M. gallisepticum Rlow, and in MRC-5 human lung fibroblasts cell culture by using relative real time reverse-transcription PCR (RT-PCR). With the exception of MGA_0338, all six genes tested (MGA_1199, 0042, 0654, 0712, 0928 and 0241) were upregulated at least at one time point during experimental infection (2-4 week post-infection). In contrast, the expression of seven out of eight IVI genes (MGA_1199, 0152, 0338, 0042, 0654, 0712, 0928) were downregulated in MRC-5 cell culture at both 2 and 4h PI; MGA_0241 was upregulated 2h PI. Our data suggest that the identified IVI antigens may have important roles in the pathogenesis of M. gallisepticum infection in vivo.
authors.affiliation: Department of Molecular Genetics and Microbiology, The Hebrew University-Haddassah Medical School, Jerusalem 91120, Israel.
authors.forename: Merav
authors.initials: M
authors.lastname: Ron
authors.affiliation: Department of Molecular Genetics and Microbiology, The Hebrew University-Haddassah Medical School, Jerusalem 91120, Israel.
authors.forename: Anna
authors.initials: A
authors.lastname: Gorelick-Ashkenazi
authors.affiliation: Mycoplasma Unit, Department of Avian and Aquatic Diseases, Kimron Veterinary Institute, Beit Dagan 50250, Israel.
authors.forename: Sharon
authors.initials: S
authors.lastname: Levisohn
authors.affiliation: Department of Clinical Microbiology and Infectious Diseases, Hadassah Hebrew University Medical Center, Ein Kerem, Jerusalem, Israel.
authors.forename: Ran
authors.initials: R
authors.lastname: Nir-Paz
authors.affiliation: Department of Pathobiology and Veterinary Science and the Center of Excellence for Vaccine Research, University of Connecticut, Storrs, CT, USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Geary
authors.affiliation: Department of Pathobiology and Veterinary Science and the Center of Excellence for Vaccine Research, University of Connecticut, Storrs, CT, USA.
authors.forename: Edan
authors.initials: E
authors.lastname: Tulman
authors.affiliation: Mycoplasma Unit, Department of Avian and Aquatic Diseases, Kimron Veterinary Institute, Beit Dagan 50250, Israel. Electronic address: innal@moag.gov.il.
authors.forename: Inna
authors.initials: I
authors.lastname: Lysnyansky
authors.affiliation: Department of Molecular Genetics and Microbiology, The Hebrew University-Haddassah Medical School, Jerusalem 91120, Israel.
authors.forename: David
authors.initials: D
authors.lastname: Yogev
mesh_terms: D000327:Adsorption; D000818:Animals; D000942:Antigens, Bacterial / Q000378:metabolism*; D002460:Cell Line; D002645:Chickens; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006801:Humans; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000235:genetics / Q000378:metabolism*; D011201:Poultry Diseases / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D037521:Virulence Factors
keywords: Immunogens; In vivo induced antigens (IVIAT); Mycoplasma gallisepticum