Novel antigenic proteins of Mycoplasma agalactiae as potential vaccine and serodiagnostic candidates

Author(s):Show less -

Barbosa, Maysa Santos ^[1] ; dos Santos Alves, Rubens Prince ^[1] ; Rezende, Izadora de Souza ^[1] ; Pereira, Samuel Santos ^[1] ; Campos, Guilherme Barreto ^{[1, 2]} ; Freitas, Leandro Martins ^[2] ; Chopra-Dewasthaly, Rohini ^[3] ; de Souza Ferreira, Luis Carlos ^[1] ; de Sa Guimara, Ana Marcia ^[1] ; Marques, Lucas Miranda ^{[1, 2]} ; Timenetsky, Jorge ^[1]

Affiliation

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, Av Prof Lineu Prestes, BR-1374 Sao Paulo - Brazil

^[2] Univ Fed Bahia, Multidisciplinary Inst Hlth, St Rio Contas 58, Vitoria Da Conquista - Brazil

^[3] Univ Vet Med Vienna, Inst Microbiol, Dept Pathobiol, Vet Pl 1, A-1210 Vienna - Austria

• Document type: Journal article
• Source: Veterinary Microbiology; v. 251, DEC 2020.
• Web of Science Citations: 0

Abstract

Contagious agalactia (CA) is a serious disease notifiable to the World Organisation for Animal Health (OIE) causing severe economic losses to sheep and goat producers worldwide. Mycoplasma agalactiae, considered as its main etiological agent, inflicts a variety of symptoms in infected animals, including keratoconjunctivitis, mastitis, arthritis, ankylosis, abortions, stillbirths and granular vulvovaginitis. Despite its significance, developing a successful vaccine remains elusive, mostly due to the lack of knowledge about M. agalactiae's pathogenicity factors and pathogenic mechanisms, including its ``core{''} antigens. The aim of this study was to identify, characterize and express antigenic proteins of M. agalactiae as potential vaccine candidates. Predicted proteins of type strain PG2 were analyzed using bioinformatic algorithms to assess their cellular localization and to identify their linear and conformational epitopes for B cells. Out of a total of 156 predicted membrane proteins, three were shortlisted as potential antigenic surface proteins, namely {[}MAG\_1560 (WP\_011949336.1), MAG\_6130 (WP\_011949770.1) and P40 (WP\_011949418.1)]. These proteins were expressed in recombinant Escherichia coli strains. Purified proteins were evaluated for their antigenicity using Western blot and ELISA using sera of M. agalactiae-naturally infected and non-infected sheep and goats. All 3 proteins were specifically recognized by the tested sera of M. agalactiae-infected animals. Also, specific rabbit antisera raised against each of these 3 proteins confirm their membrane localization using TritonX-114 phase partioning, Western and colony immunoblotting. In conclusion, our study successfully identified P40 (as proof of concept and validation) and two novel antigenic M. agalactiae proteins as potential candidates for developing effective CA vaccines. (AU)

• FAPESP's process: 14/09425-7 - Cloning, purification and characterization of recombinant antigenic proteins derived from Mycoplasma agalactiae
• Grantee: Maysa Santos Barbosa
• Support Opportunities: Scholarships in Brazil - Master