Preparation and identification of monoclonal antibodies of porcine regulatory T cells molecular markers FoxP3
Regulatory T cells(Tregs)are a type of lymphocyte that has immunomodulatory activity,and they induce autoimmune tolerance by inhibiting various effector lymphocytes.FoxP3 gene is an important regulatory gene in the differentiation,development and function main-tenance of CD4+CD25+Treg cells,and it is an important transcription factor.The combination of FoxP3 antigen on the basis of CD4+CD25+can better recognize CD4+CD25+Treg cells.In order to produce specific monoclonal antibody against porcine regulatory T cell surface marker FoxP3,a prokaryotic expression plasmid of FoxP3 epitope(1-840 bp)was constructed here by selecting a gene coding fragment with strong antigenicity.After induction and purification,BALB/c mice were immunized with the target protein,and spleen cells of the immunized mice were fused with myeloma cells,and positive hybrioma cells that could secrete antibodies stably were screened using the indirect ELISA,Western blot and indirect immunofluorescence methods.The results showed that five FoxP3 monoclonal cells with high titer(F4B9,F4A9,F1E12,F1A11 and F4A10)were finally screened and they had good specificity;all of the cells could recognize endogenous and exogenous proteins and secrete antibodies stably,which laid a foundation for qualitative or quantitative detection and verification of the function of Tregs in pigs.
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