Pe labeled cd11b

A total of 100 µL of diluted cells from the psoriasis patients (peripheral blood and
lesions from patients with erythrodermic psoriasis) and from the control group (HC
peripheral blood and non-lesions from patients with erythrodermic psoriasis) were
added to 11 separate 1.5 mL Eppendorf tubes. For both groups, one tube was used as a
negative control, and to the other 7 tubes were added 2 µL mouse anti human
FITC-labeled CD19, PE-labeled CD40, APC-labeled CD44, PE-labeled CD80, APC-labeled
CD86, PE-labeled CD11b, and APC-labeled HLA-DR monoclonal antibodies (BD Biosciences,
USA). A mixture of 2 µL FITC-labeled CD19, 2 µL PE-labeled CD40, and 2 µL APC-labeled
CD44 was added to the ninth tube. In the tenth tube, 2 µL FITC-labeled CD19, 2 µL
PE-labeled CD80, and 2 µL APC-labeled CD86 was added. In a final tube, 2 µL
FITC-labeled CD19, 2 µL PE-labeled CD11b, and 2 µL APC-labeled HLA-DR was added. The
samples were then placed at 4°C for 30 min in dark. After that, the samples were
washed with PBS twice, and centrifuged at 729 g (4°C) for 5 min
between washes. The supernatant was removed and the cell pellet was analyzed using an
ACS Calibur flow cytometer (BD Biosciences, the process strictly complied the
guidelines of the equipment).

Isolated cells were stained with FITC-labeled CD4 (BD Pharmingen, Franklin Lakes, NJ, USA; Cat. No. 553729), V500 Horizon-labeled CD4 (BD Pharmingen, Cat. No. 560783), PB-labeled CD8 (BD Pharmingen, Cat. No. 558106), PE-Cy7-labeled CD3 (BD Pharmingen, Cat. No. 560591), PE-labeled CD11b (BD Pharmingen, Cat. No. 553311), APC-labeled F4/80 (eBioscience, San Diego, CA, USA; Cat. No. 50-4801-82), FITC-labeled Ly6G (eBioscience, Cat. No. 11-5931-82), FITC-labeled CD45 (eBioscience, Cat. No. 11-0451-82), PE-labeled CD69 (eBioscience, Cat. No. 12-0691-81), APC-labeled CD86 (BD Pharmingen, Cat. No. 558703), PerCP-Cy5.5-labeled B220 (eBioscience, Cat. No. 45-0452-82), PerCP-Cy5.5-labeled IFNγ (BD Pharmingen, Cat. No. 560660), PE-Cy7 labeled TNFα (eBioscience, Cat. No. 25-7321-82), APC-labeled interleukin 4 (BD Pharmingen, Cat. No. 554436) and PE-labeled interleukin 17 (BD Pharmingen, Cat. No. 561020) antibodies.