Journal: PLoS ONE
Article Title: Murine CD4 + T Cell Responses Are Inhibited by Cytotoxic T Cell-Mediated Killing of Dendritic Cells and Are Restored by Antigen Transfer
doi: 10.1371/journal.pone.0037481
Figure Lengend Snippet: (A) C57BL/6 mice were injected with CD45.1 + WT DC or CD45.1 + WT DC loaded with OVA protein (2 mg/ml). The dLN were collected 24 h later, and the CD8 + DC, the CD8 − CD205 + skin-derived DC and the CD8 − CD205 − double negative DC populations were sorted and cultured in duplicate with CFSE-labelled, purified OT-I or OT-II T for 3 days or 5 days, respectively. OVA-specific proliferation was evaluated as CFSE dilution by flow cytometry. Each symbol represents the mean+SEM of the percentage of divided cells/well. Combined data from two independent experiments that gave similar results are shown. (B) C57BL/6 were injected with CFSE-labelled CD45-congenic OT-II CD4 + T cells and immunized 24 h later with WT or MHCII −/− DC that had been loaded with OVA protein (2 mg/ml), and treated with Ptx or left untreated. CD4 + T cell proliferation in dLN was determined by flow cytometry 3 days after DC immunization. Representative flow cytometry histograms of CD45.1 + CD4 + T cells from individual dLN are shown on the left. The mean ± SEM of the percent divided cells in each group is shown. The bar graph on the right shows mean+SEM of the number of divided CD45.1 + CD4 + T cells/dLN. Combined results from two independent experiments each with 5 mice per group are shown.
Article Snippet: To separate DC subsets, CD11c + cells were first sorted on high CD11c expression (HL3; BD Pharmingen), followed by sorting for CD205 (205yekta; eBiosciences) and CD8 (53-6.7; BD Pharmingen) using a FACSVantage SE (BD Biosciences).
Techniques: Injection, Derivative Assay, Cell Culture, Purification, Flow Cytometry
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