Protein and antigen profiles of Leptospira interrogans serovar Hardjo/ Perfil proteico e antigenico da Leptospira interrogans sorovariedade Hardjo

• Published in: Ciência rural 2009-12, Vol.39 (9), p.2539-2543
• Main Authors: Lafeta, Barbara Nobre, de Castro, Elaine Cristina, da Silva, Nivaldo
• Format: Article
• Language: eng ; spa
• Subjects: Antigens; Gram-negative bacteria; hardjoprajitno; immunomodulatory proteins; Leptospira interrogans; Membrane proteins; outer membrane protein; proteína de membrana externa; proteínas imunomodulatórias
• ISSN: 0103-8478; 1678-4596
• DOI: 10.1590/S0103-84782009000900024

The protein profile of the outer membrane of Leptospira interrogans serovar Hardjo subtype hardjoprajitno associated with the bovine natural immune response was investigated. The outer membrane proteins were extracted utilizing Triton XH4 and precipitated with acetone. The protein sample was then resolved by SDS-PAGE and reacted in western blot against sera from a hyperimmune rabbit and from naturally infected bovines. In silver stained gels, 14 protein bands were observed, among which four proteins, with 22, 29, 47 and 63kDa, appeared as major constituents. Western blot tests with hyperimmune rabbit antiserum detected bands corresponding to proteins with 35; 27; 24; 21; 17 and 14kDa, while 32kDa and 45kDa proteins were the most immunoreactive with sera from naturally infected bovines. Key words: Leptospira interrogans, outer membrane protein, hardjoprajitno, immunomodulatory proteins. Estudou-se o perfil proteico da membrana externa da Leptospira interrogans sorovariedade Hardjo, amostra hardjoprajitno, associado a resposta imune natural de bovinos infectados. Foram utilizados Triton X114 para a extracao das proteinas de membrana externa e acetona para precipita-las. As proteinas extraidas foram analisadas por SDS-PAGE e western blot contra soro de coelhos hiperimunes e de bovinos naturalmente infectados. Em geis corados com nitrato de prata, 14 bandas proteicas foram identificadas, e quatro dessas bandas, com 22, 29, 47 e 63kDa, foram as mais proeminentes. Os western blots com soro hiperimune de coelho detectaram bandas correspondentes a proteinas com pesos moleculares de 35, 27, 24, 21, 17 e 14kDa, e bandas de 32 e 45kDa destacaram-se nos testes com soros de bovinos naturalmente infectados. Palavras-chave: Leptospira interrogans, proteina de membrana externa, hardjoprajitno, proteinas imunomodulatorias.