INNATE IMMUNITY GENE EXPRESSION PROFILES AS TO THE LYMPHOID TISSUE AND CHARACTERIZATION OF THE 3’-UTR REGION OF THE GAMA INTERFERON GENE ASSOCIATED WITH ITS GENE EXPRESSION PATTERN IN THE CONJUNCTIVAL MEMBRANE OF AMAZON BUFFALOES
Bubalinoculture, inflammatory cytokines, interferon gamma, Toll-like receptors, defensins.
Buffalo production has shown high performance in recent years, especially in the Brazilian Amazon, although it presents adverse environmental conditions that influence the emergence of diseases, including eye infections. Thus, the determination of genes related to ocular innate immunity is necessary in buffalo populations. We sought to determine the expression profiles of genes related to the innate immune system in the conjunctival membrane of Amazon buffaloes and to characterize the 3’-UTR region of the Interferon gamma gene, determining its expression profile in this population. For this purpose, samples of ocular conjunctival tissue and blood were collected from buffaloes slaughtered in the State of Amapá. The animals were classified into three groups (G1, G2 and G3) according to the quantitative degree of lymphoid tissue associated with the conjunctiva by histological techniques of the conjunctival samples which were also subjected to quantification of the expression of inflammatory cytokines, toll-like receptors 4 and β-defensin-110 relative to the endogenous gene. Blood samples were subjected to conventional PCR using primers from the 3' UTR region of the Interferon gamma gene. The products were purified, sequenced and the sequences edited with corresponding ones from buffalo and mammalian species. They were aligned and compared based on genetic similarity and phylogenetic analysis. In the conjunctiva of animals from G1 there was expression only for IFNG and TLR4. While G2 there was high expression for genes IL6, IL10, IFNG, TLR4 and animals from G3 showed high expression for all genes studied. The degree of genetic similarity formed the groups of ruminants, primates, carnivores, bats, cetaceans and isolated individuals such as suiformes and rhinos. Ruminants and cetaceans have a high phylogenetic relationship while there were different evolutionary distances between buffaloes and other mammal species. Our studies served to define the innate immunological processes that occur in the conjunctival membrane of a buffalo population and to determine the evolutionary profile in relation to other mammalian species.