Fig 1: IL-17C is released basolaterally in response to HRV-1A in a dose- and replication-dependent manner. Highly differentiated HBE were treated with control medium (Con), with increasing doses of HRV-1A (106-109 HRV copies) or 109 copies of replication-deficient HRV-1A (UVHRV) for 4 h. (A) Apical wash and (B) basolateral medium were collected at the time of harvest and analyzed for IL-17C release (n = 4). Significant differences were assessed using a Kruskal-Wallis ANOVA with Dunn's multiple comparisons post-hoc test and indicated with asterisks. **p < 0.01.
Fig 2: Expression of mRNA for IL-17C, IL-17RA, and IL-17RE, and HRV1A genomic RNA in columnar and basal cell enriched populations. Highly differentiated HBE were infected with 108 copy number of HRV-1A for 24 h. Cell fractions enriched for columnar and basal cell populations were isolated as described in the legend for Figure 5 (n = 7). RNA from each subpopulation was isolated and analyzed for (A) IL-17C, (B) HRV-1A, (C) IL-17RA, and (D) IL-17RE. Significant differences were assessed using a Two-way ANOVA with Holm-Sidak's multiple comparisons post-hoc test comparing HRV-1A between cell populations and indicated with asterisks. ****p < 0.0001.
Fig 3: Cumulative basolateral IL-17C release remains significantly induced from 24 to at least 120 h following HRV-1A exposure. Highly differentiated HBE were treated apically with 108 copy number HRV-1A for 12, 24, 48, 72, 96, or 120 h and apical wash, basolateral medium, and intracellular RNA was collected at time of harvest (n = 6–8). (A) IL-17C protein was measured via ELISA in the basolateral medium. (B) Apically shed RNA and (C) cellular RNA were isolated and analyzed for HRV-1A copy number. Significant differences were assessed using a Two-way ANOVA with Holm-Sidak's multiple comparisons test at each time point and indicated with asterisks. *p < 0.05. The detection limit indicates the minimum viral copy number detected with real-time RT-PCR.
Fig 4: Basolateral IL-17C release can be induced in response to multiple serotypes of human rhinovirus as can CXCL1. Highly differentiated HBE were treated apically with major group virus HRV-16 (108 copies), minor group virus HRV-1A (108 copies), or C clade virus HRV-C15 (109 copies) for (A,C) 24 h or (B,D) 48 h and basolateral media were collected at the time of harvest (n = 4). Basolateral media were analyzed for release of IL-17C (A,B) and CXCL1 (C,D). Significant differences were assessed using a Kruskal-Wallis ANOVA with Dunnett's multiple comparisons post-hoc test and indicated with asterisks. *p < 0.05, **p < 0.01.
Fig 5: IL-17C acts in a concentration-dependent manner at the basolateral surface to induce basolateral release of CXCL1. Highly differentiated HBE were treated (A) apically or (B) basolaterally with increasing doses of exogenous IL-17C (1–100 ng/ml) for 24 h (n = 6). Apical washes and basolateral media were collected at time of harvest and analyzed for CXCL1 release. Basolateral generation of CXCL1 in response to 100 ng/ml of IL-17C applied basolaterally was significantly different than control. *p < 0.05.
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