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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 2. Periodic acid-Schiff’s (PAS) staining (a–h) and MUC5AC immunohistochemical staining (i–p) of normal human bronchial epithelial cells (NHBECs) incubated at the air–liquid interface with IL-13 (10 ng/mL) (a, b, d, f, g, i, j, l, n, o, q) or vehicle (PBS) (c, e, h, k, m, p) for 14 days in the presence of either an anti-IL-13Ra1 antibody (10 mg/mL) (b, c, j, k), an anti-IL-13Ra2 antibody (2.5 mg/mL) (d, e, l, m), an anti-IgG1 isotype control antibody (10 m g/mL) (f, n) or recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 4 mg/mL) (g, h, o, p). None of the cells were positive after treatment with IL-13 (10 ng/mL) in the absence of the first antibody (q). White arrowheads show PAS-positive goblet cells (a, d, f). Black arrowheads show MUC5AC-positive cells (i, l, n, o). The goblet cells with secretory granules stained clearly with the PAS staining method. On the other hand, there were also surface cells that did not have any goblet cell granules but were stained with PAS, PAS-positive-but-not-goblet cells. Scale bar = 100 mm.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Staining, Immunohistochemical staining, Incubation, Control, Recombinant

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 3. Number of periodic acid-Schiff’s (PAS) staining positive cells (a, PAS-positive cells), goblet cells (b), MUC5AC staining positive cells (c, MUC5AC- positive cells) and total cells (d) of normal human bronchial epithelial cells incubated with either IL-13 (10 ng/mL), IL-4 (1 ng/mL) or vehicle (PBS) for 14 days at the air–liquid interface, in the presence of either an anti-IL-13Ra1 antibody (10 mg/mL), an anti-IL-13Ra2 antibody (2.5 mg/mL), an anti-IgG1 isotype control antibody (10 mg/mL) or a recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 0.04, 0.4 or 4 mg/mL). Results are means SEM from five different bronchi. Significant differences from PBS alone are Po0.05, Po0.01 and Po0.001. Significant differences from IL-13 alone are 1Po0.05, 11Po0.01 and 111Po0.001.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Staining, Incubation, Control, Recombinant

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 4. Periodic acid-Schiff’s (PAS) staining (a, b) and MUC5AC immunohistochemical staining (c–e) of normal human bronchial epithelial cells incubated at the air–liquid interface with IL-4 (1 ng/mL) (a–e) for 14 days in the presence of recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 4 mg/ mL) (b, d). None of the cells were positive after treatment with IL-13 (10 ng/mL) in the absence of the first antibody (e). White arrowheads show PAS- positive goblet cells (a, b). Black arrowheads show MUC5AC-positive cells (c, d). Scale bar = 100 mm.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Staining, Immunohistochemical staining, Incubation, Recombinant

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 6. (a) and (b) Time course of mRNA expression of MUC5AC (a) and IL-13Ra2 (b) in normal human bronchial epithelial cells cultured with IL-13 (10 ng/mL). The results are the mean SEM from 14 different bronchi. Significant differences from values on day 0 are indicated by Po0.05, Po0.02, Po0.005.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Expressing, Cell Culture

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 5. MUC5AC protein concentrations in the supernatants of normal human bronchial epithelial cell treated with IL-13 (10 ng/mL) or IL-4 (1 ng/mL) in the presence or in the absence of either an anti-IL-13Ra1 antibody (10 mg/mL) or a recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 4 mg/mL) for 14 days. Data are expressed as percent above control vehicle (PBS). The results are mean SEM from three different bronchi. Significant differences from PBS alone are indicated by Po0.05. Significant differences from IL-13 alone are 1Po0.05.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Recombinant, Control

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 7. (a) and (b) The amount of MUC5AC mRNA expression in normal human bronchial epithelial cells (NHBECs) incubated with IL-13 (10 ng/mL) or vehicle of IL-13 (PBS) in the presence of an anti-IL-13Ra1 antibody (10 mg/mL) (a), an anti-IL-13Ra2 antibody (2.5 mg/mL) (b), an anti-IgG1 isotype control antibody (10 mg/mL) or a vehicle of antibodies (PBS) for 7 days. The results are the mean SEM from five different bronchi. Significant differences from the vehicle of IL-13 (PBS) (control) are Po0.01. Significant differences from IL-13 alone are 1Po0.05. (c) The amount of MUC5AC mRNA expression in NHBECs incubated with IL-13 (10 ng/mL) or a vehicle of IL-13 (PBS) in the presence of recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 0.04, 0.4 or 4 mg/mL) or a vehicle of IL-13Ra2 (PBS) for 7 days. The results are the mean SEM from five different bronchi. Significant differences from the vehicle of IL-13 (PBS) (control) Po0.01. Significant differences from IL-13 alone are indicated by 1Po0.05 and 11Po0.01. (d) The amount of MUC5AC mRNA expression in NHBECs incubated with IL-4 (0.1, 1 or 10 ng/mL) or a vehicle of IL-4 (PBS) in the presence of rsIL-13Ra2 (4 mg/mL) or a vehicle of IL-13Ra2 (PBS) for 7 days. The results are mean SEM from five different bronchi. Significant differences from the vehicle of IL-4 (PBS) (control) are Po0.01 and Po0.05.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Expressing, Incubation, Control, Recombinant

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 8. Immunohistochemical staining of normal human bronchial epithelial cells (NHBECs) for IL-13Ra2 proteins after exposure to PBS (a, vehicle of IL-13) or IL-13 (10 ng/mL) (b, c) for 14 days in the presence (a, b) or in the absence (c) of the first antibody. Weak staining of IL-13Ra2 protein (white arrowheads) was observed 14 days after adding the vehicle of IL-13 (PBS) (a). In contrast, the potency of IL-13Ra2 protein staining increased in NHBECs 14 days after adding IL-13 (10 ng/mL) (b) with strong staining of IL-13Ra2 protein in some cells (black arrowheads). Arrows shows goblet cells (b, c). Bar = 100 mm.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Immunohistochemical staining, Staining

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 9. (a) Time course of a soluble form of IL-13Ra2 proteins concentrations in the supernatants of normal human bronchial epithelial cells (NHBECs) cells after adding IL-13 (10 ng/mL). The results are mean SEM from four different bronchi. Significant differences from values on day 0 arePo0.05. (b) Time course of IL-13Ra2 proteins in lysates of NHBECs after adding IL-13 (10 ng/mL). The results are the mean SEM from four different bronchi. (c) A Western blot analysis of IL-13Ra2 protein in the culture supernatants (lane 1–4) and the cell lysates (lane 6) of NHBECs incubated with IL-13 (10 ng/mL) for 21 days. Lanes shows samples before (day 0, lane 1), and 7 days (day 7, lane 2), 14 days (day 14, lanes 3 and 6) and 21 days (day 21, lane 4) after adding IL-13. The recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 100 ng) is immunoblotted as a positive loading sample (lane 5). The arrow (56 kDa) shows IL-13Ra2. Data are representative of three different experiments.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Western Blot, Incubation, Recombinant

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

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 10. Western blot analysis of signal transducer and activator of transcription 6 (STAT6) (a) and the phosphorylated, active form of STAT6 (p-STAT6) (b) in the cell lysates of normal human bronchial epithelial cells incubated with IL-13 (10 ng/mL) or IL-4 (1 ng/mL) for 14 days. Lanes show the data from samples treated with either the vehicle of IL-13 or IL-4 (PBS) (lane 1), IL-13 (10 ng/mL) plus the vehicle of antibody (PBS) (lane 2), IL-13 plus anti-IL-13Ra1 antibody (10 mg/mL) (lane 3), IL-13 plus recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 4 ng/mL) (lane 4), IL-4 (1 ng/mL) plus the vehicle of antibody (PBS) (lane 5) or with IL-4 (1 ng/mL) plus rsIL-13Ra2 (4 ng/mL) (lane 6). The arrows show STAT6 (a) and p-STAT6 (b) (100120 kDa). Data are representative of three differ- ent experiments.

Article Snippet: Neutralization with anti-interleukin-13Ra1 and antiinterleukin-13Ra2 antibodies during interleukin-13 incubation To examine the blocking effect of anti-IL-13Ra1 and antiIL-13Ra2 antibodies on goblet cell hyperplasia and MUC5AC expression induced by IL-13, NHBECs were incubated with IL-13 (10 ng/mL) plus an anti-IL-13Ra1 antibody (10 mg/mL, R&D Systems Inc., Minneapolis, MN, USA), anti-IL-13Ra2 antibody (2.5 mg/mL, R&D Systems Inc.) or a vehicle (PBS) for 7 or 14 days.

Techniques: Western Blot, Incubation, Recombinant