Evaluación del factor estimulante de colonias de granulocitos y macrófagos porcino recombinante (rpGM-CSF) e interleucina -4 porcina recombinante (rpIL-4), producida en Escherichia coli

Abstract

Dendritic cells (DC) are part of the professional antigen-presenting cells (APC) capable of processing and exposing antigens in a highly specialized way. The first step is the recognition of the pathogen, which is captured to subsequently migrate to cell-rich areas. T in the lymphoid organs. DC are identified in two stages, immature located in peripheral tissues, specialized in antigen uptake and mature upregulating the porcine-specific class two major histocompatibility complex (SLA-II), CD14+, CD40+, CD80+, CD83+ and CD86+ on its surface. Porcine granulocyte-macrophage colony-stimulating factor (GM-CSF) and porcine Interleukin-4 (IL-4) are produced by activated T cells and mast cells, act as a factor of growth, differentiation, immunological regulation, proliferation, maturation and derivation of monocytes to DC (MoDC). In this study, the recombinant cytokines rpGM-CSF and rpIL-4 produced in Escherichia coli Rosetta (DE3) were synthesized with a yield of 2.354 mg for rpGMCSF and 3.17 mg for rpIL-4 from 200 mL of culture. The performance of cytokines was evaluated based on their ability to derive MoDC from monocytes isolated from peripheral blood mononuclear cells (PBMC). Obtaining MoDC with a phenotype MHC-II+, CD14+ and low CD80 and dendritic morphology, determined by flow cytometry. Additionally, the phagocytic capacity of MoDC was analyzed using rhodamine-labeled LPS (E. coli), evidencing interaction between cells and pathogens, which could potentially induce an adaptive-type immune response.