Journal: PLoS ONE
Article Title: Toll-Like Receptor Ligands Induce Expression of the Costimulatory Molecule CD155 on Antigen-Presenting Cells
doi: 10.1371/journal.pone.0054406
Figure Lengend Snippet: (A-D and F-H) Cd155 −/− mice (n = 12) and WT littermate (n = 12) were immunized i.p. with 20 µg OVA and 50 µg ODN 1668, 20 µg OVA or saline alone. One week later the immunization was repeated. 21 days after first immunization, serum levels of OVA-specific IgG2a/c (A), total IgG (B) and IgG1 (C) antibodies in OVA- and CpG-immunized Cd155 −/− mice (open squares) and WT littermates (closed squares) were determined by indirect ELISA at a 1∶100,000 serum dilution. Combined data of three independent experiments are shown. Groups shown in (A-C) were compared using Student’s t-test. Means ± SEM. are indicated, * p <0.05. (D) Splenic B cells (CD19 + cells) of saline, OVA and OVA + CpG ODN 1668-injected WT mice were analyzed for CD155 expression (thick line) by flow cytometry. Filled histogram shows B cells stained with isotype control. (E) Splenocytes of WT mice (n = 3) were stained for CD3, CD4, CD44, CD25, TIGIT and DNAM-1 expression. Dot blots show TIGIT and DNAM-1 expression in splenocytes electronically gated on CD3 + CD4 + CD25 − CD44 low (R1, naïve phenotype, middle panel) or CD3 + CD4 + CD44 high (R2, effector/memory phenotype, right panel) cells. Left panel illustrates CD25 and CD44 gating strategy used for R1 and R2. (F) 5×10 6 splenocytes of mice injected with OVA and CpG were cultured for 48 hrs, after which the culture supernatant was analyzed for IL-4 levels by ELISA. Data are representative of two independent experiments. (G and H) CD3 + CD4 + splenocytes of OVA- and CpG-immunized WT and Cd155 −/− mice (n = 7) were analyzed for intracellular IL-4 (G) and GATA-3 (H) expression by flow cytometry. Data presented in (F-G) were analyzed using Student’s t-test and means ± SEM are shown. ** p <0.01, ns indicates not significant.
Article Snippet: 5×10 6 spleen cells of OVA and CpG immunized mice were cultured in 1 ml RPMI medium (Invitrogen, Singapore) for 48 hrs after which, the levels of IL-4, IFN-γ, IL-12p70 and IL-10 in the supernatant was measured by ELISA according to the manufacturer’s instructions (eBioscience, USA).
Techniques: Indirect ELISA, Injection, Expressing, Flow Cytometry, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay
R&D Systems is a verified supplier 
R&D Systems manufactures this product 
