Journal: The Journal of infectious diseases

Article Title: Sensitized lymphocytes and CD40 ligation augment interleukin-12 production by human dendritic cells in response to Toxoplasma gondii.

doi: 10.1086/314601

Figure Lengend Snippet: Figure 2. Presence of lymphocytes augments interleukin (IL)-12 pro- duction by generated DC in response to T. gondii. DC (50,000/well, open bars) either in absence or presence of lymphocytes from same donor (ratio 1:10, 500,000 lymphocytes, solid bars; ratio 1:4, 200,000 lymphocytes, hatched bars) were incubated at 377C with either live tachyzoites (toxo; E:T ratio 1:5, 10,000 PLK strain; E:T ratio 1:2, 25,000 PLK strain) or Toxoplasma lysate antigen (TLA; 2.5 or 0.5 mg/ mL) for 24 h. Lipopolysaccharide (LPS) (100 ng/mL) was included as positive stimulus for IL-12 induction. Lymphocytes alone were used at highest concentration (500,000/well, dotted bars). Cell supernatants were harvested, and production of IL-12 p40 subunit was measured by ELISA. Data are expressed in pg/mL for 3 wells/ mean 5 SD experiment.

Article Snippet: Two IL-12 ELISA systems were used within these studies: Quantikine human IL-12 p40 subunit immunoassay kits (R&D Systems) and IL-12 p70 DUOSET ELISA (Genzyme, Cambridge, MA).

Techniques: Generated, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Synovial fibroblasts directly induce Th17 pathogenicity via the cyclooxygenase/prostaglandin E2 pathway, independent of IL-23.

doi: 10.4049/jimmunol.1300274

Figure Lengend Snippet: FIGURE 1. Limited contribution of IL-1b and IL-6 to the perseverance of the proinflammatory loop induced upon Th17–RASF interaction. Sorted primary memory CCR62 T cells and/or CCR6+ T cells (Th17) from healthy individuals were stimulated with anti-CD3/anti-CD28 and cultured with RASF for 2 d. (A) IL-1b, IL-6, and RORC gene transcription levels were detected by quantitative real-time PCR, and IL-6 and IL-17A protein levels were detected by ELISA in CCR62 T cell–RASF cultures (filled bars) or in Th17–RASF cultures (open bars). Gene transcription levels were normalized against HPRT values. (B) Th17–RASF cultures were stim- ulated with anti-CD3/anti-CD28 and cultured for 2 d in the presence or absence of anti–IL-1b (2.5 mg/ml) and/or anti–IL-6R (25 mg/ml). Ex- pression of indicated cytokines and MMPs was determined by ELISA in supernatant of Th17–RASF cultures in absence (black bars) or presence of anti–IL-1b (dark gray bars), anti–IL-6R (light gray bars), or both (white bars). (C) IL-23p19 and IL-12/23p40 gene transcription levels were detected by quantitative real-time PCR and normalized against HPRT gene expression values. Means and SEM are given for six healthy donors. Results are representative of at least three independent experiments. *p , 0.05, **p , 0.01, ***p , 0.001. n.d., Not detectable.

Article Snippet: IL-23 expression was detected by ELISA using the combination of biotin-conjugated anti-human IL-12/23p40 (eBioscience) and anti-human IL-23p19 (R&D Systems).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Gene Expression