Development And Characterization Of Monoclonal Antibodies Against Chicken CD3 And IL-2

Chicken CD3 is one of the most important cell surface markers of T cells. During antigen recognition, CD3 complex are involved in signal transduction, which ultimately leads to T-lymphocyte activation after the initial recognition steps. Anti-chicken CD3 monoclonal antibodies (McAbs) are useful tools to study the functions of T lymphocytes expressing CD3 molecules and cellular immunity in chickens. Chicken interleukin-2 (ChIL-2) is a glycoprotein produced principally by activated T cells and some B cells. It plays key roles in T cell growth and differentiation, B cell development and activation of NK cells. It has the homologous biology activity as a molecule of immune modulation. Anti-chicken IL-2 McAb can be used to a simple, rapid and highly reliable bioassay for the detection of ChIL-2, and it could be used to study the structure and function of ChIL-2 and its immunobiology in the avian immune system. In this study, the recombinant plasmids pcDNA3.1-IL-2 and pcDNA3.1-CD3 were used to immunize the six-week-old BALB/c mice respectively, after fusing and selecting, the McAbs against ChIL-2 and chicken CD3 were obtained and some of their characteristics were identified. The results provide helpful information for developing the methods and techniques for the detection and analysis of immune cells and immune regulations in chicken model.1. Construction and expression of recombinant plasmid pcDNA3.1-IL-2Chicken IL-2 cDNA was amplified by PCR and subcloned into eukaryotic plasmid pcDNA3.1(+) to construct recombinant plasmid pcDNA3.1-IL-2. The recombinant plasmid was identified by restriction enzyme digestion and then transfected into COS-7 cells. The results of indirect immunofluorescent assay (IFA) revealed the protein of...