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In asthma we see in vivo evidence of GC-resistant pathways in airway smooth muscle (ASM) bundles that can be modeled in vitro by exposing cultured ASM cells to TNF-\u03b1\/IFN-\u03b3. This action drives GC insensitivity via protein phosphatase 5\u2013dependent impairment of GC receptor phosphorylation. In this study, we investigated whether KCa3.1 ion channels modulate the activity of GC-resistant pathways using our ASM model of GC insensitivity. Immunohistochemical staining of endobronchial biopsies revealed that KCa3.1 channels are localized to the plasma membrane and nucleus of ASM in both healthy controls and asthmatic patients, irrespective of disease severity. Western blot assays and immunofluorescence staining confirmed the nuclear localization of KCa3.1 channels in ASM cells. The functional importance of KCa3.1 channels in the regulation of GC-resistant chemokines induced by TNF-\u03b1\/IFN-\u03b3 was assessed using complementary inhibitory strategies, including KCa3.1 blockers (TRAM-34 and ICA-17043) or KCa3.1-specific small hairpin RNA delivered by adenoviruses. KCa3.1 channel blockade led to a significant reduction of fluticasone-resistant CX3CL1, CCL5, and CCL11 gene and protein expression. KCa3.1 channel blockade also restored fluticasone-induced GC receptor-\u03b1 phosphorylation at Ser211 and transactivation properties via the suppression of cytokine-induced protein phosphatase 5 expression. The effect of KCa3.1 blockade was evident in ASM cells from both healthy controls and asthmatic subjects. In summary, KCa3.1 channels contribute to the regulation of GC-resistant inflammatory pathways in ASM cells: blocking KCa3.1 channels may enhance corticosteroid activity in severe asthma.<\/jats:p>","DOI":"10.4049\/jimmunol.1300104","type":"journal-article","created":{"date-parts":[[2013,8,1]],"date-time":"2013-08-01T02:13:50Z","timestamp":1375323230000},"page":"2624-2636","source":"Crossref","is-referenced-by-count":29,"title":["Functional KCa3.1 Channels Regulate Steroid Insensitivity in Bronchial Smooth Muscle Cells"],"prefix":"10.1093","volume":"191","author":[{"given":"Latifa","family":"Chachi","sequence":"first","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aarti","family":"Shikotra","sequence":"additional","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S Mark","family":"Duffy","sequence":"additional","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Omar","family":"Tliba","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, School of Pharmacy, Thomas Jefferson University , Philadelphia, PA 19107"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Brightling","sequence":"additional","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Bradding","sequence":"additional","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yassine","family":"Amrani","sequence":"additional","affiliation":[{"name":"Department of Infection, Immunity and Inflammation, University of Leicester , Leicester LE1 7RH,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2013,9,1]]},"reference":[{"key":"2025032405255254100_r1","doi-asserted-by":"crossref","first-page":"2341","DOI":"10.1164\/ajrccm.162.6.ats9-00","article-title":"Proceedings of the ATS workshop on refractory asthma: current understanding, recommendations, and unanswered questions","volume":"162","author":"American Thoracic Society","year":"2000","journal-title":"Am. 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