title: Identification of vaccine candidate antigens of Staphylococcus pseudintermedius by whole proteome characterization and serological proteomic analyses.
abstract: The recent emergence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) has complicated considerably the treatment of infections caused by these bacteria. Therefore new treatment strategies are urgently needed, namely through the development of vaccines towards the control of bacterial infections. Our study describes an extensive characterization of the proteome of S. pseudintermedius through a 2-DE MALDI-TOF/TOF approach, followed by SERological Proteome Analysis (SERPA) to identify potential vaccine candidate antigens. We were able to identify 361 unique proteins, of which 39 are surface proteins. In order to assess the immunogenic potential of S. pseudintermedius proteins, a Western blot analysis of two-dimensional gels was carried out with serum from healthy dogs, dogs with atopic dermatitis infected and not infected with S. pseudintermedius. Only immunogenic areas detected by ≥ 50% of the dogs with atopic dermatitis infected with S. pseudintermedius sera and by <50% of the healthy dogs sera were excised and identified from Coomassie-colloidal stained gels. The areas identified by IgE were not considered as vaccine targets, because those proteins could induce hypersensitivity. We were able to identify 13 unique proteins after in-gel digestion of selected protein gel spots, with 4 antigenic proteins showing promising features for vaccine development. No specific antibodies were identified in the dogs with atopic dermatitis not infected with S. pseudintermedius sera that could contribute to prevention of infection. The SERPA approach employed in this study revealed novel candidate therapeutic targets for the control of S. pseudintermedius infections.
authors.affiliation: Antibiotic Resistance Laboratory, CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (FMV-UL), Av. da Universidade Técnica, 1300-477 Lisbon, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Natacha
authors.initials: N
authors.lastname: Couto
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Martins
authors.affiliation: Dermatology and Immunoalergology Service, Clinical Research, CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (FMV-UL), Av. da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Ana Mafalda
authors.initials: AM
authors.lastname: Lourenço
authors.affiliation: Antibiotic Resistance Laboratory, CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (FMV-UL), Av. da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Constança
authors.initials: C
authors.lastname: Pomba
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal. Electronic address: varela@itqb.unl.pt.
authors.forename: Ana
authors.initials: A
authors.lastname: Varela Coelho
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004283:Dog Diseases; D004285:Dogs; D020543:Proteome / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011711:Pyoderma / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D013203:Staphylococcal Infections / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D013209:Staphylococcal Vaccines / Q000235:genetics / Q000276:immunology; D013210:Staphylococcus / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D020543:Proteome; D013209:Staphylococcal Vaccines
keywords: MRSP; Proteomics; S. pseudintermedius; SERPA; Vaccine candidates