Fitc rat anti mouse cd4

Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples of psoriatic patients and healthy donors by Ficoll-Paque gradient centrifugation (17144003, Cytiva). Human CD4⁺ T cells were isolated using a CD4⁺ T cell isolation kit (Cat#480,009, BioLegend) following the manufacturer’s protocol. Peripheral blood was obtained from experimental mice, and mouse CD4⁺ T cells were isolated using a CD4⁺ T cell isolation kit (Cat#19,852, Stem cell) following the manufacturer’s protocol. The human and mouse CD4⁺ T cell purity was confirmed to be above 95% by flow cytometric analysis. Flow cytometry was performed using FACS Calibur (BD Biosciences) and Cytoflex (Beckman) devices. Data were analyzed using FlowJo software (Tree Star, Ashland, OR). Th17 cells were defined as CD4⁺ IL17⁺ cells, and Tregs were defined as CD4⁺CD25⁺ Foxp3⁺ cells.
The following antibodies were used: FITC rat anti-human CD4 (Biolegend), APC mouse anti-human CD4 (Biolegend), PE/Cyanine5 mouse anti-human CD25 (Biolegend), PE mouse anti-human IL-17 A (eBioscience), FITC rat anti-mouse CD4 (Biolegend), PE rat anti-mouse CD3 (Biolegend), PE rat anti-mouse IL-17 A (Biolegend), and APC rat anti-mouse CD25 (Biolegend).

PLGA (lactic acid: glycolide 65:35, molecular weight (Mw) = 40,000–75,000), chitosan (Mw = 50,000–190,000 Da), poly vinyl alcohol (PVA, Mw = 31,000–50,000), concanavalin A, and complete Freund’s adjuvant were purchased from Sigma-Aldrich (St. Louis, MO, USA).
We purchased the Sodium tripolyphosphate (Mw = 367.86) from Aladdin and Enhanced Cell Counting Kit-8 from Beyotime. Likewise, the BCA Protein Assay Kit was acquired from CW Biotech, China.
The Roswell Park Memorial Institute medium (RPMI 1640), heat-inactivated fetal bovine serum (FBS), and penicillin–streptomycin solutions were purchased Gibco. Antibodies PE rat anti-mouse CD86 (clone: GL1), PE rat anti-mouse CD83 (clone: Michel-19), APC hamster anti-mouse CD11c (clone: HL3), APC hamster anti-mouse CD3e (clone: 145-2C11), FITC rat anti-mouse CD4 (clone: RM4-5), and FITC rat anti-mouse CD8α (clone: 53-6.7) were purchased from Bio-legend. The purified recombinant proteins of HcARF1 and pET-32a expressed in E. coli [5 (link)] were obtained from Molecular Parasitology and Immunology laboratory at Nanjing Agricultural University.