Determinación de la proteína C reactiva en especies de animales con diferentes patologías

Abstract

[ES] Las enfermedades que cursan con un proceso inflamatorio (infección, trauma, heridas¿) están asociadas a signos fisiológicos característicos y, a cambios en ciertas proteínas séricas, conocidas como proteínas de fase aguda. Entre ellas, se encuentra la proteína C reactiva (C-RP) que es un potente activador de las rutas clásica y alternativa del sistema del complemento y, participa además, en otros procesos biológicos e inmunológicos. Esta proteína pertenece a la familia de las pentraxinas y se encuentra tanto en invertebrados como en vertebrados. Actualmente, hay disponible una amplia bibliografía sobre la C-RP en humanos y en algunas especies animales como perros y gatos; sin embargo, es escasa la información en pequeños mamíferos (hurones, cobayas, conejos, etc.) y, especialmente, en aves. Por ello, se considera de interés el estudio de la C-RP en estas especies. Las técnicas empleadas en este trabajo de investigación se basan, fundamentalmente, en ensayos cromatográficos e inmunoensayos, aplicados a muestras procedentes de animales sanos y de animales con diferentes patologías.

[EN] The acute phase response (APR) is the reaction that occurs in the body in response to changes in hemostasis caused by infection, trauma, neoplastic growth or immune disorders. The APR includes changes in the concentrations of certain proteins present in plasma called acute phase proteins (APPs). Pentraxins are a superfamily of proteins found within APPs. There is a protein belonging to this group is very well studied and characterized in man. This protein called C-reactive protein (CRP), because it has the peculiar property of precipitating the polysaccharide C, a component of the bacterial wall. The CRP is widely accepted as a biomarker for cardiovascular disease and inflammation. Generally, the plasma levels are below 2.0 mg / L to healthy individuals, but CRP concentration can rise up to 1000 times in inflammation conditions. This has enabled their clinical application as a diagnostic tool. In this work several techniques for possible application in the detection and quantification of CRP in different animal species, such as the Western Blot (WB), indirect competitive ELISA format and antibody microarrays are used. In the WB, the CRP is detected in samples from mammals with a band of 26 kDa. However, in samples from birds and reptiles, this band does not appear. Therefore, for detection it could readjust the amount of protein and antibody or perform the technique using as a primary antibody anti-CRP of bird or reptile. In this case, there would be purified CRP of these animals and obtaining an antibody from the purified CRP. As for the ELISA conducted to quantify samples an increasing gradient with increasing amount of primary antibody is observed, but the variation gradient of the concentration of free CRP is observed in wells. Finally, the antibody microarray technique shows that human anti-CRP antibody recognized with higher affinity the CRP from other species. So, the optimized human CRP assay is suboptimal for quantifying the samples and is necessary to have patterns of CRP from other animal species, to develop specific assays for each animal species. In addition, the CRP is not sufficiently conserved between species as to use a human anti-CRP antibody for specific recognition of CRP in different animal species.