Anti mhcii pe

Chitosan, with an MW of 600 kDa and a deacetylation degree of 85%, was procured from Sigma Chemical Co. (St Louis, MO, USA). Polyvinyl alcohol (PVA) with a purity of 98.0–98.8% was purchased from Fisher Scientific Co., Ltd. (Brussels, Belgium); DMEM (high glucose) complete medium (4.5 g/L D-glucose, sodium pyruvate, L-glutamine, phenol red, 10% FBS (0500), and 1% penicillin/streptomycin) was purchased from Shanghai Zhongqiaoxinzhou Biotechnology Co., Ltd. (Shanghai, China); The CCK-8 kit was purchased from Biyuntian Biotechnology Co., Ltd. (Shanghai, China); PLGA (MW = 10,000–20,000), pancreatin, and FITC were purchased from Shanghai Yuanye Biology Co., Ltd. (Shanghai, China); fluorescently labeled anti-mouse monoclonal antibodies (anti-CD80-PE, anti-CD40-FITC, anti-CD86-PE, anti-MHCII-PE, anti-CD4-FITC, anti-CD3-Percp, and anti-CD8-PE) were obtained from Thermo Fisher Scientific (Asheville, NC, USA).

Cells were stained with anti-CD4-FITC (rat IgG2b,κ), anti-CD25-PE (rat IgG1,λ), anti-CD206-PE (rat IgG2b,κ), anti-CD40-FITC (Armenian hamster IgM,κ), anti-F4/80-PerCP-Cy5.5 (rat IgG2a,κ), anti-MHC II-PE (rat IgG2b,κ), anti-CD86-PE-Cy5 (rat IgG2a,κ), anti-CD11b-PE-Cy5 (rat IgG2b,κ), anti-CD11c-PE (Armenian hamster IgG), anti-IFN-γ-PE (rat IgG1,κ), anti-IL-17-PerCP-Cy5.5 (rat IgG2a), anti-FoxP3-PE-Cy5 (rat IgG2a,κ) antibodies (all from Thermo Fisher Scientific, Waltham, MA, USA). Appropriate isotype control antibodies were used where necessary to set gates for cell marker positivity. Typically, proportion of isotype control antibody-stained cells was <1%. Cells were analyzed with a CyFlow Space flow cytometer (Partec, Munster, Germany). Results of cytofluorometry analysis are presented as proportion of cells bound by an appropriate antibody or as calculated absolute cell numbers per organ. Gating strategies for Th1, Th17 and Treg are presented in Figure 1.