Mouse anti bovine cd4 fitc

Antibody staining of cells and flow cytometric quantification of T and B cells were described in detail by Liermann et al. (28 (link)). Briefly, cells were stained by monoclonal antibodies for cluster of differentiation (CD)2 (T cells) (mouse anti-bovine CD2: FITC; Bio-Rad Laboratories, Hercules, USA), CD4 (T helper cells) (mouse anti-bovine CD4: FITC; Bio-Rad), CD8 (cytotoxic cells) (mouse anti-bovine CD8: Alexa Fluor^® 647; Bio-Rad), and CD21 (B cells) (mouse anti-bovine CD21: RPE, Bio-Rad) or the corresponding isotype controls (mouse IgG1 negative control: FITC; mouse IgG2a negative control: FITC; mouse IgG2a negative control: Alexa Fluor^® 647; mouse IgG2b negative control: RPE; Bio-Rad). A flow cytometer Type GalliosTM (Beckman Coulter GmbH, Krefeld Germany) was used.

For phenotyping of lymphocytes, EDTA blood samples were double-stained with monoclonal antibodies for CD4+ (mouse anti-bovine CD4+:FITC (fluorescein isothiocyanate); Bio-Rad, Hercules, CA, USA) and CD8+ (mouse anti-bovine CD8+:PE; Bio-Rad Hercules, CA, USA) or their related isotype controls (mouse IgG2a negative control RPE, and mouse IgG2b: FITC negative control; Bio-Rad, Hercules, CA, USA). Further procedure was conducted as described above and as described in [20 (link)].