cd4 dc isolation kit  (Miltenyi Biotec)

Journal: Cellular and Molecular Immunology

Article Title: Deficiency of the G protein Gαq ameliorates experimental autoimmune encephalomyelitis with impaired DC-derived IL-6 production and Th17 differentiation

doi: 10.1038/cmi.2016.65

Figure Lengend Snippet: Gαq-KO inhibits in vivo Th17 development. Leukocytes were isolated from the lymph nodes of WT mice and Gαq-KO mice on day 10 PI and analyzed with flow cytometry. (a–c) Th1 and Th17 cells were analyzed by intracellular staining of IFN-γ and IL-17A, respectively, in the CD4+ gate. (d–f) Lymph node leukocytes from EAE-induced WT mice and Gαq-KO mice were re-stimulated in vitro with MOG35–55 (20 μg/ml) for 72 h, and IL-17A (d), IFN-γ (e) and TGF-β (f) in supernatants were analyzed with ELISA. (g) qPCR analysis of Th1- and Th17-related gene expression in lymph nodes. The data are expressed as the mean±s.e.m. (n=3), * P<0.05 versus WT-EAE.

Article Snippet: DC-T cell co-culture A CD4 + CD62L + T cell Isolation Kit II (Miltenyi Biotec, Auburn, CA, USA) was used to isolate naive T cells from single-cell suspensions from the spleen; in the first step, non-CD4 + cells were magnetically depleted, and magnetically labeled CD62L + T cells were positively selected in the second step.

Techniques: In Vivo, Isolation, Flow Cytometry, Staining, In Vitro, Enzyme-linked Immunosorbent Assay, Gene Expression

Journal: Cellular and Molecular Immunology

Article Title: Deficiency of the G protein Gαq ameliorates experimental autoimmune encephalomyelitis with impaired DC-derived IL-6 production and Th17 differentiation

doi: 10.1038/cmi.2016.65

Figure Lengend Snippet: Gαq-KO inhibits in vitro Th17 differentiation. (a–f) Naive CD4+ T cells from WT and Gαq-KO mice were induced to differentiate into Th1 (a, b) or Th17 (d, e) cells in vitro. Antigen-specific Th1 and Th17 responses were measured by ELISA for IFN-γ and IL-17A production in the supernatant (c, f). The data are expressed as the mean±s.e.m. (n=3), ***P<0.001 versus WT.

Article Snippet: DC-T cell co-culture A CD4 + CD62L + T cell Isolation Kit II (Miltenyi Biotec, Auburn, CA, USA) was used to isolate naive T cells from single-cell suspensions from the spleen; in the first step, non-CD4 + cells were magnetically depleted, and magnetically labeled CD62L + T cells were positively selected in the second step.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay