HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

Epitope mapping of Mycoplasma hyopneumoniae using phage displayed peptide libraries and the immune responses of the selected phagotopes.

Abstract
Phage display techniques have been widely employed to map the epitope structures which served as the basis for developing molecular vaccines. In the present study, we applied this technique to map the epitopes of Mycoplasma hyopneumoniae, the etiologic agent causing swine enzootic pneumonia, and evaluated directly the immune responses in mice of the selected phage-displayed epitopes (phagotopes). Two phage-displayed random peptide libraries were biopanned with the protein A-purified IgG of the rabbit anti-M. hyopneumoniae hyperimmune serum and the selected phage clones were sequenced and analyzed. Some of the inserts of the selected phagotopes showed a good match with the known proteins of M. hyopneumoniae. Others, which did not match with any known proteins, but shared extensive homology with each other, were clustered and classified as the conformational epitopes of M. hyopneumoniae. To evaluate the potential of using these phagotopes as effective vaccines, several phage clones were chosen to immunize mice. IgA coproantibody, IgA in bronchoalveolar lavage fluid and serum IgG responses were assayed. The serum raised by the phage clones clearly recognized several major mycoplasmal proteins indicating that the phagotope-induced immune responses were antigen-specific. The stronger IgG1 response revealed that the immune responses of the epitope-displaying phage were mainly through Th2 activation. The growth inhibition assay showed that the selected phage clones CS4 and varphi58 are potential vaccine candidates and suggested that the mycoplasmal 97 kDa, 56 kDa, 30 kDa and 23 kDa proteins may play important roles in the immune responses. The present work demonstrates that the whole epitope profile of a microorganism can be obtained through screening the phage displayed peptide libraries with the hyperimmune serum and reveals the potential of using epitope-displaying phages as peptide vaccines.
AuthorsWen-Jen Yang, Jen-Feng Lai, Kou-Cheng Peng, Huey-Jenn Chiang, Chun-Nan Weng, David Shiuan
JournalJournal of immunological methods (J Immunol Methods) Vol. 304 Issue 1-2 Pg. 15-29 (Sep 2005) ISSN: 0022-1759 [Print] Netherlands
PMID16054642 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Adhesins, Bacterial
  • Antibodies, Bacterial
  • Antibodies, Viral
  • Bacterial Vaccines
  • Epitopes
  • Growth Inhibitors
  • Immunoglobulin G
  • Peptide Library
Topics
  • Adhesins, Bacterial (genetics, immunology)
  • Administration, Intranasal
  • Amino Acid Sequence
  • Animals
  • Antibodies, Bacterial (biosynthesis)
  • Antibodies, Viral (biosynthesis)
  • Bacterial Vaccines (administration & dosage, immunology)
  • Bacteriophage M13 (growth & development, immunology)
  • Epitope Mapping (methods)
  • Epitopes (immunology)
  • Female
  • Growth Inhibitors (administration & dosage, immunology)
  • Immunoglobulin G (biosynthesis)
  • Injections, Intraperitoneal
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Mycoplasma (immunology)
  • Peptide Library
  • Rabbits
  • Swine

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: