il-17af  (Meso Scale Diagnostics LLC)

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein Suppresses the Th17 Response Indirectly by Attenuating the Antigen Presentation Ability of Monocyte Derived Dendritic Cells in Experimental Autoimmune Encephalomyelitis

doi: 10.3389/fimmu.2021.589200

Figure Lengend Snippet: The Th17 responses are suppressed by one-dose CRP injection in WT EAE mice.Splenocytes were isolated form EAE and EAE CRP mice, then re-stimulated with 50 ug/ml MOG 24 h for qPCR, 72 h for ELISA and FACS. (A) Th17 relevant cytokine (IL-17) and transcription factor (ROR γ t) were examined by qPCR and ELISA (n = 8). (B) Th1 relevant cytokine (IFN- γ ) and transcription factor (T-bet) were measured by qPCR and ELISA (n = 8). (C) Th2 relevant cytokine (IL-4) and transcription factor (GATA-3) were detected by qPCR and ELISA (n = 8). (D) Th2 relevant cytokine (IL-10) and transcription factor (Foxp3) were tested by qPCR and ELISA (n = 8). (E) Th17 (CD4 + IL-17 + ) and Th1 (CD4 + IFN- γ + ) cells were examined by FACS (n = 3). Data are presented as mean ± SEM; p < 0.05 was considered statistically significant.

Article Snippet: Anti-mouse CD25 APC (Cat: 17-02510-82, Lot: 4276862), anti-mouse MHC-II APC(I-A/I-E) (Cat: 17-5321-81, Lot: 1991457), anti-mouse CD86 APC (Cat: 17-0862-81, Lot: 1984132), anti-CD3 mAb (Cat: 16-0031-85, Lot: 4349473), anti-CD28 mAb (Cat: 16-0281-85, Lot: 1974623), antibodies to ROR γ t (Cat: 14-6981-82, Lot: 1936480), T-bet (Cat: 14-5825-82, Lot: 2012147) and Mouse IL-17AF ELISA Set (Cat: 88-8711-88, Lot: 4291151) were obtained from ebioscience (San Diego, CA, USA).

Techniques: Injection, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein Suppresses the Th17 Response Indirectly by Attenuating the Antigen Presentation Ability of Monocyte Derived Dendritic Cells in Experimental Autoimmune Encephalomyelitis

doi: 10.3389/fimmu.2021.589200

Figure Lengend Snippet: CRP reduces Th17 responses in splenocytes from WT mice.Splenocytes and CD4 + T cells were isolated from WT mice to distinguish the indirect and direct regulations of CRP on Th17. (A) The protein expression levels of IL-17 and IFN- γ were measured with CRP treatment in both splenocytes and CD4 + T cells by ELISA (n = 6). (B) The mRNA expression levels of IL-17 and IFN- γ were measured with CRP treatment in both splenocytes and CD4 + T cells by qPCR (n = 4). (C) Splenocytes and CD4 + T cells were cultured with CRP in Th17 and Th1 polarization conditions, then ROR γ t, T-bet, p-STAT-3, and p-STAT-1 were examined by WB. (D) The quantitative and statistical analysis of the WB results was presented (n = 3). Data are presented as mean ± SEM; p < 0.05 was considered statistically significant.

Article Snippet: Anti-mouse CD25 APC (Cat: 17-02510-82, Lot: 4276862), anti-mouse MHC-II APC(I-A/I-E) (Cat: 17-5321-81, Lot: 1991457), anti-mouse CD86 APC (Cat: 17-0862-81, Lot: 1984132), anti-CD3 mAb (Cat: 16-0031-85, Lot: 4349473), anti-CD28 mAb (Cat: 16-0281-85, Lot: 1974623), antibodies to ROR γ t (Cat: 14-6981-82, Lot: 1936480), T-bet (Cat: 14-5825-82, Lot: 2012147) and Mouse IL-17AF ELISA Set (Cat: 88-8711-88, Lot: 4291151) were obtained from ebioscience (San Diego, CA, USA).

Techniques: Isolation, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein Suppresses the Th17 Response Indirectly by Attenuating the Antigen Presentation Ability of Monocyte Derived Dendritic Cells in Experimental Autoimmune Encephalomyelitis

doi: 10.3389/fimmu.2021.589200

Figure Lengend Snippet: The decrease in Th17 response disappeared in Fc γ R2B −/− EAE mice with CRP injection. (A) qPCR and ELISA analysis of IL-17 and ROR γ t expression in splenocytes and CD4 + T cells from Fc γ R2B −/− EAE mice and Fc γ R2B −/− EAE CRP (n = 8). (B) qPCR and ELISA analysis of IFN- γ and T-bet expression in splenocytes and CD4 + T cells from Fc γ R2B −/− EAE mice and Fc γ R2B −/− EAE CRP (n = 8). (C) qPCR and ELISA analysis of IL-4 and GATA-3 expression in splenocytes and CD4 + T cells from Fc γ R2B −/− EAE mice and Fc γ R2B −/− EAE CRP (n = 8). (D) qPCR and ELISA analysis of IL-10 and Foxp3 expression in splenocytes and CD4 + T cells from Fc γ R2B −/− EAE mice and Fc γ R2B −/− EAE CRP (n = 8). (E) Flow cytometry analysis of Th17 (CD4 + IL-17 + ) and Th1 (CD4 + IFN- γ + ) cells by cell surface and intercellular staining from Fc γ R2B −/− EAE mice and Fc γ R2B −/− EAE CRP (n = 3). Data are presented as mean ± SEM; p < 0.05 was considered statistically significant.

Article Snippet: Anti-mouse CD25 APC (Cat: 17-02510-82, Lot: 4276862), anti-mouse MHC-II APC(I-A/I-E) (Cat: 17-5321-81, Lot: 1991457), anti-mouse CD86 APC (Cat: 17-0862-81, Lot: 1984132), anti-CD3 mAb (Cat: 16-0031-85, Lot: 4349473), anti-CD28 mAb (Cat: 16-0281-85, Lot: 1974623), antibodies to ROR γ t (Cat: 14-6981-82, Lot: 1936480), T-bet (Cat: 14-5825-82, Lot: 2012147) and Mouse IL-17AF ELISA Set (Cat: 88-8711-88, Lot: 4291151) were obtained from ebioscience (San Diego, CA, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Staining