V500 conjugated anti cd45

Flow cytometric analysis was performed using blood samples collected before treatment (weeks 7–8 post-immunization) and at 2, 3, 5, 6, 7, and 8 weeks after treatment (weeks 9–10, 10–11, 13, 14, 15, and 16 post-immunization). All antibodies were purchased from BD Biosciences (San Jose, CA, USA). For analysis of T-cell populations, peripheral blood mononuclear cells (PBMCs) were stained with V500–conjugated anti-CD45 (BD Biosciences), PE-Cy7–conjugated anti-CD3, PerCP-Cy5.5–conjugated anti-CD4, APC-Cy7–conjugated anti-CD8, PE–conjugated anti-CD28, and APC–conjugated anti-CD95 antibodies. For analysis of B-cell populations, PBMCs were stained with V500–conjugated CD45, PE-Cy7–conjugated anti-CD3, FITC–conjugated anti-CD20, V450–conjugated anti-CD27, PerCP-Cy5.5–conjugated anti-CD21, PE–conjugated anti-IgD, and APC–conjugated anti-IgM antibodies. For analysis of regulatory T cells, PBMCs were stained with V500–conjugated anti-CD45, FITC–conjugated anti-CD4, PE-Cy7–conjugated anti-CD25, and PE–conjugated anti-FoxP3 antibodies. Flow cytometric analysis was carried out using a BD LSRFORTESSA cell analyzer (BD Biosciences).

The V450-conjugated anti-CD3, Alexa Fluor®700-conjugated anti-CD3, BV650-conjugated anti-CD4, FITC-conjugated anti-CD4, BV605-conjugated anti-CD11b, BV421-conjugated anti-CD11c, BV650-conjugated anti-CD14, PE-Cy^TM7-conjugated anti-CD14, Alexa Fluor®700-conjugated anti-CD16, BV421-conjugated anti-CD16, PE-CF594-conjugated anti-CD16, phycoerythrin (PE)-conjugated anti-CD19, Alexa Fluor®700-conjugated anti-CD19, Alexa Fluor®700-conjugated anti-CD20, V500-conjugated anti-CD45, BV786-conjugated anti-NKp46, Alexa Fluor®700-conjugated anti-CD56, PE-conjugated anti-CD64, PE-Cy^TM7-conjugated anti-CD86, PerCP-Cy5.5-conjugated anti-CD123, BV711-conjugated anti-CD141, BV786-conjugated anti-PDL2, and PE-CF594-conjugated anti-CD163 antibodies were from BD Biosciences. The APC-Alexa Fluor®750-conjugated anti-HLA-DR antibody was purchased from Beckman Coulter. The PE-conjugated anti-CD1c, APC-conjugated anti-CD303, and APC-conjugated anti-SLAN antibodies were from Miltenyi Biotech. The FITC-conjugated anti-CD192 antibody was purchased from R&D Systems. All antibodies were used according to their manufacturer's recommendations. Antibodies combination for monocytes: CD3, CD11b, CD14, CD16, CD19, CD45, CD56, CD64, CD163, CD192, NKp46, HLA-DR, SLAN, and DC: CD1c, CD3, CD4, CD11b, CD11c, CD14, CD16, CD19, CD20, CD45, CD56, CD86, CD123, CD141, CD303, PDL2, HLA-DR.