Comparative Immunology, Microbiology and Infectious Diseases 35 (2012) 209–216 Contents lists available at SciVerse ScienceDirect Comparative Immunology, Microbiology and Infectious Diseases j o ur nal homep age : w ww.elsevier.com/locate/cimid Immunological characterization of Mycoplasma hyopneumoniae recombinant proteins Simone Simionatto a,b , Silvana B. Marchioro a , Vanessa Galli a , Clarice B. Brum a , Catia S. Klein c , Raquel Rebelatto c , Everton F. Silva a , Sibele Borsuk a , Fabricio R. Conceic ¸ ão a , Odir A. Dellagostin a,∗ a Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil b Faculdade de Ciências Biológicas e Ambientais, Universidade Federal da Grande Dourados, Dourados, MS, Brazil c Embrapa Suínos e Aves, Concórdia, SC, Brazil a r t i c l e i n f o Article history: Received 15 July 2011 Received in revised form 18 November 2011 Accepted 9 January 2012 Keywords: Enzootic pneumonia Recombinant protein Vaccine a b s t r a c t Mycoplasma hyopneumoniae, the primary pathogen of enzootic pneumonia, is highly preva- lent worldwide and causes major economic losses to the pig industry. Commercial vaccines are widely used in the control of this disease, however, they provide only partial protec- tion. The aim of this study was to evaluate 34 recombinant proteins of M. hyopneumoniae expressed in Escherichia coli. Antigenic and immunogenic properties of these proteins were analyzed. For this, the proteins were tested against hyperimmune and convalescent pig sera through ELISA and Western blot. Immunogenicity of the recombinant proteins was evaluated in BALB/c mice following intramuscular inoculation. Most antigens were able to induce a strong immune response and sera from inoculated mice were able to recognize native proteins by cell ELISA and Western blot. Several recombinant proteins were specif- ically recognized by convalescent pig sera, indicating they are expressed during infection. These data may help to develop more efficacious vaccines against M. hyopneumoniae. © 2012 Elsevier Ltd. All rights reserved. 1. Introduction Mycoplasma hyopneumoniae is the primary pathogen of enzootic pneumonia (EP), a chronic respiratory disease in pigs prevalent in almost all swine producing areas. The dis- ease causes significant economic losses due to increased medication use and decreased performance of the pigs [1]. Moreover, infected animals are more sensitive to concur- rent respiratory infections by other pathogenic bacterial species or viruses [2]. ∗ Corresponding author at: Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Campus Universitário S/N, CEP 96010- 900 Pelotas, RS, Brazil. Tel.: +55 53 3275 7350; fax: +55 53 3275 7551. E-mail addresses: simonesimionatto@ufgd.edu.br (S. Simionatto), odir@ufpel.edu.br (O.A. Dellagostin). Control of M. hyopneumoniae infection can be accom- plished by a number of different ways, namely by optimization of management practices and housing conditions, the use of antimicrobials and vaccination. Commercial vaccines, consisting of inactivated whole-cell preparations, are widely used to control M. hyopneumo- niae infection [3]. The main effects of vaccination include less clinical symptoms, lung lesions and medication use, and improved performance. However, these vaccines are not enough to eliminate M. hyopneumoniae from infected pig herds [4]. Investigation of new vaccines is actively occurring, especially recombinant subunit and naked DNA vaccines [4–15]. These strategies have the potential to result in a new vaccine able to control M. hyopneumoniae infection. However, only three of these vaccines have been tested in pigs and were able to provide partial protection in vaccinated animals [10,14,15]. Thus, the identification of 0147-9571/$ – see front matter © 2012 Elsevier Ltd. All rights reserved. doi:10.1016/j.cimid.2012.01.007