quantikine human vap 1 elisa kit  (R&D Systems)

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

Article Title: Restriction of de novo pyrimidine biosynthesis inhibits Th1 cell activation and promotes Th2 cell differentiation.

doi: 10.4049/jimmunol.169.6.3392

Figure Lengend Snippet: FIGURE 6. Leflunomide increases Th2 effector functions in vivo. Six- week-old C57BL/6 mice were fed 1% CMC (; n 5) or 5 mg/kg le- flunomide dissolved in 1% CMC/day (f; n 5) and immunized with KLH as described in Materials and Methods. Sera were collected 1 wk after the secondary immunization, and the levels of total IgE and KLH-specific total IgG, IgG1, and IgG2a were determined by ELISA. A, Leflunomide inhib- ited KLH-specific IgG production. The mean KLH-specific IgG levels of the control mice were defined as 100 relative units. B, Leflunomide sig- nificantly increases the level of Th2-dependent KLH-specific IgG1. The mean KLH-specific IgG1 levels of the control mice were defined as 100 relative units. C, The level of Th1-dependent, KLH-specific IgG2a Abs decreases in leflunomide-treated animals. The mean KLH-specific IgG2a levels of the control mice were defined as 100 relative units. D, Lefluno- mide shifts the balance of Th2-dependent KLH-specific IgG1/Th1-depen- dent KLH-specific IgG2a toward IgG1. E, Leflunomide did not signifi- cantly alter the total production of IgE. , Significant differences from control cultures (p 0.05).

Article Snippet: Detection of cytokine secretion from splenocytes by ELISA Splenocytes (1 106/ml) from mice that were immunized with KLH and treated or untreated with leflunomide were stimulated with KLH (10 g/ ml) and anti-CD28 (5 g/ml) or PMA (1 mM) and ionomycin (20 ng/ml) for 72 h. IL-4 and IFN- were assessed in the culture supernatants by high sensitive Quantikine ELISA kits (R&D Systems) according to the manufacturer’s instructions.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Inhibition, Control