Table 1.
Patient details (FEV1: forced exhaled volume over 1 second; FCV: forced vital capacity; NA: not available).
Fig 1.
Treatment of hBEC with EP 7630 and RV16 infection.
Table 2.
Antibodies used for protein analysis.
Abcam, Cambridge, U.K.
Fig 2.
(A) Concentration-dependent preventive effect of EPs 7630 on hBEC number at 2 days after RV16 infection. (B) Concentration dependent reduction of RV16 positive (green) hBEC after 2 days. Bars represent mean ± S.E.M. of six cell lines in each group. Statistics were calculated as Student’s paired t-test. * indicates P<0.05 compared to untreated hBEC (control) and **indicate p<0.05 compared to RV16 infected cells. Insert: PCR product for RV-16 in epithelial cells in the presence and absence of EPs 7630 over 24 hours. Complete data of survival and RV-positive cells is provided in the supporting information (Figs A and B in S1 File, Tables A and B in S2 File).
Fig 3.
Effect of EPs 7630 on ICOS (A) and ICOSL (B) expression by human primary bronchial epithelial cells following 1 day incubation. Bars represent OD. mean ± S.E.M.of six independent experiments in different cells lines of each group. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. Representative Immuno-blots are presented as inserts to Fig 3A and 3B and complete data of immuno-blots are shown in the supporting information (Tables A and B and Fig C in S3 File).
Fig 4.
(A) Representative immune-cytochemistry of C1qR localisation in the absence and presence of EPs 7630 or RV16 infection in primary human hBEC of healthy controls. Similar results were obtained in hBEC obtained from patients with moderate-severe asthma or COPD. (B) Semi-quantitative image analysis (ImageJ) of C1qR in six Western-blots in each donor group. Bars represent OD. mean ± S.E.M.of six independent experiments in different cells lines of each group. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. Representative Immuno-blots are presented as inserts to Fig 4A and 4B and complete data of immuno-blots are shown in the supporting information (Table A and Fig B in S4 File).
Fig 5.
(A) One day incubation with EPs 7630 stimulated the expression of the anti-viral cell surface protein β-defensin-1. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. (B) Representative immunofluorescence photographs of β-defensin-1 expression in primary human epithelial cells in the presence and absence of EPs 7630 at three days in primary human hBEC of healthy controls. Similar results were obtained in hBEC obtained from patients with moderate-severe asthma or COPD. Bars represent OD. mean ± S.E.M.of six independent experiments in different cells lines of each group. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. Representative Immuno-blots are presented as inserts to Fig 5A and 5B and complete data of immuno-blots are shown in the supporting information (Table A and Fig B in S5 File).
Fig 6.
(A) Concentration dependent induction of SOCS-1 by EPs 7630 in the presence and absence of RV16 infection. Bars represent mean ± S.E.M. of six cell lines in each group. “C” indicates untreated hBEC; * indicate P-value <0.05 compared to control; ** indicates P-value < 0.05 compared to RV16 infected hBEC. Data of immuno-blot analysis are presented in supporting information (S6 File). (B) Representative immunofluorescence image of SOCS1 at 48 hours in the presence and absence of EPs 7630 in primary human hBEC of healthy controls. Similar results were obtained in hBEC obtained from patients with moderate-severe asthma or COPD.