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

Article Title: IL-16 promotes leukotriene C(4) and IL-4 release from human eosinophils via CD4- and autocrine CCR3-chemokine-mediated signaling.

doi: 10.4049/jimmunol.168.9.4756

Figure Lengend Snippet: FIGURE 3. Receptors involved in IL-16-induced activation of eosino- phils. A, CD4 mediates the activation of eosinophils by IL-16. As indi- cated, eosinophils were pretreated for 30 min with inhibitors of IL-16/CD4 interaction (including sCD4 and Fab of OKT4 anti ()-CD4 Ab (or irrel- evant control W6/32 anti-HLA Ab)) or with the Gi protein inhibitor PTX and then were incubated with IL-16 (100 nM) for 1 h for induction of lipid bodies, priming for enhanced LTC4 production (fluid-phase assays), and IL-4 release (solid-phase matrix assays). B, Involvement of Gi protein- coupled CCR3 receptors. As indicated, eosinophils were pretreated for 30 min with antagonists of PAF (CV6209 and BN52021) and CCR3 (Met- RANTES (MetR) and anti-CCR3 neutralizing mAb (or isotype control rat IgG2a)) receptors and then were activated with IL-16. Results are means SD from three independent assays. and , p 0.05 compared with nonstimulated and IL-16-stimulated eosinophils, respectively. B, Values represent the calculated percentage of inhibition with antagonists in com- parison with stimulated increases in lipid body numbers, LTC4 production, and IL-4 release above baselines.

Article Snippet: Eosinophils were stimulated with IL-16 (0.01–100 nM; R&D Systems), RANTES, or eotaxin (6 nM; R&D Systems) for time periods ranging from 5 min to 3 h. For inhibitor studies, cells were pretreated for 30 min with pertussis toxin (PTX) (20 ng/ml; Calbiochem, La Jolla, CA), recombinant soluble CD4 (sCD4) (50 ng/ml; R&D Systems), Fab of anti-CD4 (clone OKT4; American Type Culture Collection, Manassas, VA) and anti-HLA class I (clone W6/32; Sigma-Aldrich) mAbs (1 g/ml), anti-CCR3 mAb (clone 61828.111; R&D Systems), or isotype control rat IgG2a at 10 g/ml (BD PharMingen, San Diego, CA), met-RANTES (60 nM; R&D Systems), the platelet-activating factor (PAF) receptor antagonists CV6209 and BN52021 (10 M; Biomol, Plymouth Meeting, PA), brefeldin A (BFA) (0.1 and 1 g/ml; Biomol), or their vehicles, as indicated.

Techniques: Activation Assay, Control, Incubation, Inhibition

Journal:

Article Title: Expression of CCL28 by Reed-Sternberg Cells Defines a Major Subtype of Classical Hodgkin's Disease with Frequent Infiltration of Eosinophils and/or Plasma Cells

doi:

Figure Lengend Snippet: Summary of the Primers Used for RT-PCR

Article Snippet: Cells were washed twice with phosphate-buffered saline containing 3% fetal bovine serum and incubated at 4°C for 30 minutes with the following monoclonal or polyclonal antibodies; mouse anti-human CXCR4 (12G5; DAKO Japan, Kyoto, Japan), mouse anti-human CXCR5 (51505.111; DAKO Japan), mouse anti-human CXCR6 (56811.111; R&D Systems, Minneapolis, MN), mouse anti-human CCR4 (53103.111; BD Biosciences, Mountain View, CA), mouse anti-human CCR6 (53103.111; R&D Systems), mouse anti-human CCR7 (2H7; BD Biosciences), rat anti-human CCR3 (61828; R&D Systems), goat anti-human CCR8 (Alexis Biochemicals, San Diego, CA), and goat anti-human CXCL16 (R&D Systems).

Techniques: