Caracterização de Leptospira spp. quanto à presença e conservação dos genes da família lig: importantes alvos para utilização em vacina e testes de diagnóstico

Abstract

Pathogenic Leptospira species contain one or more genes of the lig family, which code for Lig-family proteins. Lig proteins ( Leptospiral immunoglobulin-like proteins ) are considered important virulence factors, and they may be involved in tissue penetration and adhesion to host cells. Lig proteins have been implicated as the main factors involved in pathogenesis of Leptospira spp. Three genes, ligA, ligB and ligC are known, which code for proteins with the same name. The increasing interest in the characterization of the genes code for Lig proteins is due to their strong immunoprotective and diagnosis potential. Thus, it was necessary to evaluate the degree of conservation among Lig genes and proteins in order to predict their usefulness as antigens for the induction of a broad range protection. Four of the pathogenic species of Leptospira had the lig genes sequenced in this study, comprising L. interrogans, L. noguchii, L. weilli and L. borgpetersenii. Among the LigB proteins sequenced so far, LigB from L. interrogans Copenhageni FIOCRUZ L1-130 appears as the most conserved when compared to LigB from other species. The sequencing and comparative analysis of lig genes and proteins demonstrated a low identity among the different genomospecies of Leptospira spp. The alignment of lig genes sequences from all the genomospecies enabled us to design primers that allowed the amplification of a fragment from each gene. Such approach demonstrated that ligB gene is present in all the pathogenic species, while ligA gene is only found in L. interrogans and L. kirschneri and ligC gene is present in L. interrogans, L. kirschneri, L. weilli and some serovars of L. noguchii. The sequencing of the amplified fragment, derived from 33 serovars which comprise 6 genomospecies, and their comparative analysis together with the corresponding region from ligB sequences available in GenBank, enabled a phylogenetic analysis. It was possible to differentiate pathogenic species based on the DNA sequence of this ligB fragment.