Mouse anti chicken cd3 pe

Peripheral blood samples were collected from the wing veins of 10 birds in each group at 0, 7, 14, 21 and 28 dpv. Peripheral blood lymphocytes were isolated from the peripheral blood samples. The samples were diluted with equal volumes of sterile PBS, overlaid onto chicken peripheral blood lymphocyte isolation liquid (Solarbio, Beijing, China) and centrifuged at 500 g for 20 min at room temperature. The peripheral blood lymphocytes at the interface were collected and washed with sterile PBS. A final volume of 0.1 mL was stained with 3 µL of Mouse Anti-Chicken CD3-PE, 2 µL of Mouse Anti-Chicken CD4-PE/CY7 and 2 µL of Mouse Anti-Chicken CD8α-FITC antibodies (SouthernBiotech, Birmingham, AL, USA), respectively. The concentrations of the Mouse Anti-Chicken CD3-PE, CD4-PE/CY7 and CD8α-FITC antibodies were adjusted to 0.3, 0.1, and 1 µg/0.1 mL, respectively. All the samples were assessed for the percentages of CD3⁺, CD4⁺ and CD8⁺ T lymphocytes using a BD AccuriTMC6 Flow Cytometer (Becton, Dickinson and Company, Minneapolis, MN, USA). All the staining reactions were conducted according to the manufacturer’s protocols.