Specific binding of anti-chCD205 and internalization of antigen delivery complex. A The recombinant parts of chicken CD205 receptor used for monoclonal antibody preparation and the monoclonal antibody characterization. The CD205B and CD205F proteins were purified from inclusion bodies on DEAE Sepharose and separated on Tris-Tricine polyacrylamide gel stained by Coomassie blue (left). Monoclonal mouse anti-CD205 antibodies were analyzed using Western blots. Results of two representative detections of monoclonal anti-CD205 specific for CD205B or CD205F are shown. B CD205 expression on leukocytes. Splenocytes and peripheral leukocytes were stained with anti-chicken CD205 antibody and antibodies recognizing each of the indicated antigens. Histograms represent the CD205 fluorescence for cells gated for positive expression of the markers indicated at the left. The results of one representative of three experiments are shown. C Median fluorescence intensity (MFI) of CD205 stained cells and standard deviation. D Internalization of streptavidin-CD205-FITC complex by chicken primary splenocytes one (left) and two (right) hours of incubation in different temperatures. Dotted line plot non-labelled cells (biot—control IgG); Black line plot—FITC labelled cells incubated in 4 °C; Gray line plot—FITC labelled cells incubated in 40 °C.