P phycoerythrin conjugated anti cd8

Venous blood samples were collected from all patients (except one male patient who did not offer a blood sample). The proportions of T lymphocyte subsets (e.g., CD3⁺, CD4⁺, CD8⁺ T cells, and NK cells) were examined by a BD FACS CantoTM II flow cytometer (BD Bioscience, CA, United States; previously described in PMID: 8404602). The blood samples were stained with antibodies as follows: Pcy5-conjugated anti-CD3, FITC-conjugated anti-CD4, P-phycoerythrin-conjugated anti-CD8 (BD Biosciences, CA, United States), and PE-conjugated anti-CD16/CD56 (Beckman Coulter, CA, United States). These tests were all carried out in triplicate using the same batch of kits.

Venous blood samples (1 mL) were collected from individual patients and healthy individuals immediately after visiting the hospital. To determine the frequency of T cells (CD3/CD4/CD8), natural killer (NK) cells, and monocytes (CD14), blood samples were stained with the following antibodies: Pcy5-conjugated anti-CD3, FITC-conjugated anti-CD4, P-phycoerythrin-conjugated anti-CD8 (BD Biosciences, CA, USA), and PE-conjugated anti-CD16/CD56 (Beckman Coulter, CA, USA).
The frequency of different types of immunocompetent cells was characterized by flow cytometry as previously described (PMID: 8404602). Briefly, at least 1 × 10⁵ cells were analyzed using BD FACS Canto^TM II flow cytometer (BD Bioscience, CA, USA). The staining of T cells or monocytes was performed with a single antibody or a combination of the following antibodies: anti-Tim-3-APC (R&D Systems), anti-PD-1-PE (eBiosciences), anti-PD-L1-BrilliantViolet421 (Biolegend), and anti-PD-L2-APC (Biolegend). The isotype-matched immunoglobulins were used as controls. The tests were performed at least in triplicate.