easystep mouse t cell enrichment kits  (STEMCELL Technologies Inc)

Journal: Frontiers in Immunology

Article Title: Multiple Regulatory Levels of Growth Arrest-Specific 6 in Mucosal Immunity Against an Oral Pathogen

doi: 10.3389/fimmu.2018.01374

Figure Lengend Snippet: Diminished pathogen-specific adaptive immunity in Gas6 −/− mice. (A) Schematic presentation of the murine periodontitis model used in this study. Antibiotic pre-treated mice were infected via oral gavage, three times at 2-day intervals, with 1 × 10 10 CFU of Porphyromonas gingivalis strain 53977 (Pg). Six weeks after the last inoculation, the mice were analyzed. (B) IFN-γ production by restimulated splenocytes quantified by ELISA representing the mean value ± SEM ( n = 8). (C) Pg-specific IgG titers measured by ELISA in the plasma of the mice, representing the mean of OD 450 values ± SEM ( n = 8). (D–F) Flow cytometry analysis of the percentages of CD45 + cells (D) , CD4 + T cells (E) , and FOXP3-expressing CD4 + T regulatory cells (F) in the gingival tissues of infected mice ( n = 5). Representative data of one out of three independent experiments is shown.

Article Snippet: OT-I CD8 + T cells or OT-II CD4 + T cells were purified by negative selection with the EasyStep mouse CD8 + or CD4 + T-cell enrichment kits, respectively, according to the manufacturer’s instructions (StemCell Technologies, Canada).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: Multiple Regulatory Levels of Growth Arrest-Specific 6 in Mucosal Immunity Against an Oral Pathogen

doi: 10.3389/fimmu.2018.01374

Figure Lengend Snippet: Growth arrest-specific 6 (GAS6) inhibits dendritic cell (DC) maturation and limits T-cell activation by DCs. (A) BMDCs generated from WT and Gas6 −/− mice were stimulated with LPS for 24 h. Representative FACS histograms and graphs demonstrating the expression and mean florescence intensity (MFI) of MHCII (top) and CD86 (bottom) on BMDCs, respectively. Gray histograms represent staining on MHC-negative cells in the culture. (B) DCs purified from cervical lymph nodes (LNs) of Gas6 −/− and WT mice were stimulated with LPS for 24 h. Representative FACS plots demonstrating the expression of CD86, MHCII, and CCR7 are presented. (C,D) DCs enriched from cervical LNs of Gas6 −/− and WT mice were stimulated with LPS and OVA peptides and then co-cultured with naive CFSE-labeled OT-II CD4 + T cells or OT-I CD8 + T cells. (C) Three days post-stimulation the dilution in CFSE levels on the T cells was analyzed by flow cytometry, representative FACS histograms are presented, numbers indicate the mean of three repeats per group ± SEM. (D) Bar graphs show the concentrations of IFN-γ secreted to the supernatant by activated CD8 + T cells and represent the mean of four repeats per group ± SEM. Representative data of one out of two independent experiments is shown.

Article Snippet: OT-I CD8 + T cells or OT-II CD4 + T cells were purified by negative selection with the EasyStep mouse CD8 + or CD4 + T-cell enrichment kits, respectively, according to the manufacturer’s instructions (StemCell Technologies, Canada).

Techniques: Activation Assay, Generated, Expressing, Staining, Purification, Cell Culture, Labeling, Flow Cytometry