ABSTRACT
Chagas disease, which is caused by the parasite Trypanosoma cruzi, is an important cause of cardiomyopathy in Latin America. It is estimated that 10%-30% of all infected individuals will acquire chronic chagasic cardiomyopathy (CCC). The etiology of CCC is multifactorial and involves parasite genotype, host genetic polymorphisms, immune response, signaling pathways and autoimmune progression. Herein we verified the impact of the recombinant form of P21 (rP21), a secreted T. cruzi protein involved in host cell invasion, on progression of inflammatory process in a polyester sponge-induced inflammation model. Results indicated that rP21 can recruit immune cells induce myeloperoxidase and IL-4 production and decrease blood vessels formation compared to controls in vitro and in vivo. In conclusion, T. cruzi P21 may be a potential target for the development of P21 antagonist compounds to treat chagasic cardiomyopathy.
Subject(s)
Cardiomyopathies/etiology , Chagas Disease/pathology , Protozoan Proteins/antagonists & inhibitors , Trypanosoma cruzi/metabolism , Animals , Cardiomyopathies/drug therapy , Cell Adhesion/drug effects , Cell Line , Cell Survival/drug effects , Chagas Disease/drug therapy , Chagas Disease/parasitology , Chemotaxis/drug effects , Cytokines/metabolism , Disease Models, Animal , Inflammation/chemically induced , Inflammation/metabolism , Inflammation/prevention & control , Interleukin-4/metabolism , Leukocytes/cytology , Leukocytes/immunology , Leukocytes/metabolism , Male , Mice , Mice, Inbred C57BL , Neovascularization, Physiologic/drug effects , Peroxidase/metabolism , Protozoan Proteins/genetics , Protozoan Proteins/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/pharmacology , Recombinant Proteins/therapeutic use , Trypanosoma cruzi/isolation & purificationABSTRACT
Structural studies of proteins normally require large quantities of pure material that can only be obtained through heterologous expression systems and recombinant technique. In these procedures, large amounts of expressed protein are often found in the insoluble fraction, making protein purification from the soluble fraction inefficient, laborious, and costly. Usually, protein refolding is avoided due to a lack of experimental assays that can validate correct folding and that can compare the conformational population to that of the soluble fraction. Herein, we propose a validation method using simple and rapid 1D (1)H nuclear magnetic resonance (NMR) spectra that can efficiently compare protein samples, including individual information of the environment of each proton in the structure.
