Mycoplasma gallisepticum in vivo induced antigens expressed during infection in chickens

Merav Ron, Anna Gorelick-Ashkenazi, Sharon Levisohn, Ran Nir-Paz, Steven J. Geary, Edan Tulman, Inna Lysnyansky*, David Yogev

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

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.

Original languageAmerican English
Pages (from-to)265-274
Number of pages10
JournalVeterinary Microbiology
Volume175
Issue number2-4
DOIs
StatePublished - 2015

Bibliographical note

Funding Information:
This research was supported by Research Grant Number 2007474 from the United States-Israel Binational Science Foundation (BSF).

Publisher Copyright:
© 2014 Elsevier B.V.

Keywords

  • Immunogens
  • In vivo induced antigens (IVIAT)
  • Mycoplasma gallisepticum

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