Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: Comparison of expression of IL-5 (a) and IL-1β (b) mRNAs using RT-PCR and Southern blot analysis in human ASM cells after 0-, 3-, 6-, and 24-hour exposure to human control serum and human atopic asthmatic serum. Expression of RPL7 mRNA was used to control for gel loading. The blots were probed with human specific IL-5, IL-1β, and RPL7 32P-labeled cDNA probes (see Methods). Whereas mRNA expression of IL-5 or IL-1β was unaltered in cells that were exposed to control serum, the mRNA signals for IL-5 and IL-1β were upregulated in cells that were exposed to atopic asthmatic serum. The temporal patterns of enhanced mRNA expression of these cytokines were characterized by an initial (at 3 hours) increased expression of IL-5 mRNA, followed (at 6 hours) by an increased mRNA expression of IL-1β. In contrast, mRNA expression of the constitutively expressed RPL7 gene was unaltered.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Comparison, Expressing, Reverse Transcription Polymerase Chain Reaction, Southern Blot, Labeling

Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: Comparisons of the release of IL-5 and IL-1β proteins into the culture media of human ASM cells after 0-, 3-, 6-, and 24-hour exposure to human control serum (open symbols) and human atopic asthmatic serum (filled symbols). In contrast to control serum that had no appreciable effect, ASM cells exposed to human atopic asthmatic serum showed significantly increased levels of both IL-5 (filled circles) and IL-1β (filled squares) protein release into the cell culture media. Moreover, the temporal pattern for IL-5 release appeared relatively earlier (at 3 hours) than that for IL-1β release. Data represent mean ± SE values.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Cell Culture

Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: (a) Comparison of human IL-1β mRNA expression examined using RT-PCR and Southern blot analysis in cultured human ASM cells after 0-, 3-, 6-, and 24-hour exposure to SFM alone (control) or IL-5. Relative to the unaltered constitutively expressed RPL7 mRNA signal, IL-1β mRNA expression was markedly upregulated at 3 and 6 hours in the IL-5–exposed ASM cells, whereas the IL-1β mRNA signal was virtually undetectable in the control cells. (b) Western blot analyses of human IL-1β expression in membrane/lysate samples isolated from cultured human ASM cells and tissues after 24-hour exposure to SFM alone and to IL-5. In contrast to control cells and tissues maintained in SFM alone, the cells and tissues treated with IL-5 exhibited markedly upregulated expression of IL-1β protein.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Comparison, Expressing, Reverse Transcription Polymerase Chain Reaction, Southern Blot, Cell Culture, Western Blot, Membrane, Isolation

Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: Comparison of human IL-1β protein accumulation in the culture media of human ASM cells after 0-, 3-, 6-, and 24-hour exposure to SFM alone (control; open bars) or IL-5 (filled bars). Relative to unaltered release of IL-1β protein in control cells, ASM cells exposed to IL-5 exhibited significantly increased levels of IL-1β protein release into the cell culture media. Data represent mean ± SE values.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Comparison, Cell Culture

Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: Comparison of contractile dose-response relationships to ACh in paired rabbit ASM segments after treatment with SFM alone (open circles) or with a maximum effective concentration of IL-5 (5 ng/mL) in the absence (filled circles) or presence (open squares) of IL-1ra. Relative to ASM exposed to SFM alone (control), the heightened constrictor responses to ACh in the IL-5–treated tissues were prevented by cotreatment of tissues with IL-1ra. Data represent mean ± SE values from 6 paired experiments.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Comparison, Concentration Assay

Journal:

Article Title: Autocrine interaction between IL-5 and IL-1? mediates altered responsiveness of atopic asthmatic sensitized airway smooth muscle

doi:

Figure Lengend Snippet: Comparison of relaxation dose-response relationships to isoproterenol in paired rabbit ASM segments half-maximally contracted with their respective ED50 doses of ACh after treatment with SFM alone (open circles) or with IL-5 in the absence (filled circles) or presence (open squares) of IL-1ra. Relative to ASM exposed to SFM alone (control), the attenuated relaxation responses to isoproterenol in the IL-5–treated tissues were prevented by cotreatment of tissues with IL-1ra. Data represent mean ± SE values from 6 paired experiments.

Article Snippet: The IL-1 receptor antagonist, anti-human IL-5 receptor and IL-4 neutralizing antibodies, IL-5 and IL-1β ELISA kits, mouse anti-human IL-1β primary antibody, and anti-mouse secondary antibody used in protein assay studies were purchased from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Comparison

Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

Article Title: Prolonged allergen challenge in mice leads to persistent airway remodelling

doi:

Figure Lengend Snippet: Effect of prolonged ovalbumin (OVA) challenge on cytokine levels. IL-4 (a, b), IL-5 (c, d), IL-13 (e, f) and IFN-γ (g, h) levels were measured in bronchoalveolar lavage (left panel) and lung tissue homogenates (right panel) by ELISA in samples from alum controls, acute and chronic phase killed 24 h after the final OVA challenge and mice from day 80. Data are expressed as mean ± SEM; n = 4–17/group and *P<0.05 in comparison with alum control mice. ND, non-detectable.

Article Snippet: Paired antibodies for murine IL-4 and IFN-γ (PharMingen, Oxford, UK), IL-5 (Endogen-Pierce, Tattenhall, UK), TGF-β1 and eotaxin/CCL11 (R&D Systems, Abingdon, UK) were used in standardized sandwich ELISAs according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Elevated IL-5 levels in pleural fluid of patients with paragonimiasis westermani

doi: 10.1046/j.1365-2249.2001.01425.x

Figure Lengend Snippet: Individual IL-5 levels in pleural fluid in various diseases. PW, Paragonimiasis westermani. IL-5 concentrations in PW were significantly higher than those in other disease groups (P < 0·0001). Means are indicated by horizontal bars. Detection limit of IL-5 is represented as a horizontal dotted line (< 20 pg/ml).

Article Snippet: Briefly, 50 μl/well MoAb to human IL-5 (TRFK5; PharMingen, San Diego, CA) (4 μg/ml NaHCO 3 , pH 8·2) was bound to microtitre plates by incubating at 4°C overnight.

Techniques:

Journal:

Article Title: Elevated IL-5 levels in pleural fluid of patients with paragonimiasis westermani

doi: 10.1046/j.1365-2249.2001.01425.x

Figure Lengend Snippet: (a) Relationship between percentages of eosinophils in peripheral blood and IL-5 concentrations in pleural fluid of patients with paragonimiasis westermani (n = 11). (b) Relationship between percentages of eosinophils and IL-5 concentrations in pleural fluid of patients with paragonimiasis westermani (n = 10).

Article Snippet: Briefly, 50 μl/well MoAb to human IL-5 (TRFK5; PharMingen, San Diego, CA) (4 μg/ml NaHCO 3 , pH 8·2) was bound to microtitre plates by incubating at 4°C overnight.

Techniques: