Phosphatidylinositol 3-Kinase Is Required for Rhinovirus-induced Airway Epithelial Cell Interleukin-8 Expression

Abstract

Rhinovirus (RV) is a common cause of asthma exacerbations. The signaling mechanisms regulating RV-induced airway epithelial cell responses have not been well studied. We examined the role of phosphatidylinositol (PI) 3-kinase in RV-induced interleukin (IL)-8 expression. Infection of 16HBE14o- human bronchial epithelial cells with RV39 induced rapid activation of PI 3-kinase and phosphorylation of Akt, a downstream effector of PI 3-kinase. RV39 also colocalized with cit-Akt-PH, a citrogen-tagged fluorescent fusion protein encoding the pleckstrin homology domain of Akt, indicating that 3-phosphorylated PI accumulates at the site of RV infection. Inhibition of PI 3-kinase and Akt attenuated RV39-induced NF-κB transactivation and IL-8 expression. Inhibition of PI 3-kinase also blocked internalization of labeled RV39 into 16HBE14o- cells, suggesting that the requirement of PI 3-kinase for RV39-induced IL-8 expression, at least in part, relates to its role in viral endocytosis. Rhinovirus (RV) is a common cause of asthma exacerbations. The signaling mechanisms regulating RV-induced airway epithelial cell responses have not been well studied. We examined the role of phosphatidylinositol (PI) 3-kinase in RV-induced interleukin (IL)-8 expression. Infection of 16HBE14o- human bronchial epithelial cells with RV39 induced rapid activation of PI 3-kinase and phosphorylation of Akt, a downstream effector of PI 3-kinase. RV39 also colocalized with cit-Akt-PH, a citrogen-tagged fluorescent fusion protein encoding the pleckstrin homology domain of Akt, indicating that 3-phosphorylated PI accumulates at the site of RV infection. Inhibition of PI 3-kinase and Akt attenuated RV39-induced NF-κB transactivation and IL-8 expression. Inhibition of PI 3-kinase also blocked internalization of labeled RV39 into 16HBE14o- cells, suggesting that the requirement of PI 3-kinase for RV39-induced IL-8 expression, at least in part, relates to its role in viral endocytosis. Rhinovirus (RV) 2The abbreviations used are: RV, rhinovirus; ANOVA, analysis of variance; COPD, chronic obstructive pulmonary disease; ELISA, enzyme-linked immunosorbent assay; ICAM, intercellular adhesion molecule; IL, interleukin; IκB, inhibitor of κB; MOI, multiplicity of infection; NF-κB, nuclear factor-κB; PH, pleckstrin homology; PI, phosphatidylinositol; PI(4,5)P2, PI 4,5-bisphosphate; PI(3,4,5)P3, PI 3,4,5-trisphosphate; PBS, phosphate-buffered saline; TCID, tissue culture infectivity dose; TNF, tumor necrosis factor; IKK, IκB kinase. 2The abbreviations used are: RV, rhinovirus; ANOVA, analysis of variance; COPD, chronic obstructive pulmonary disease; ELISA, enzyme-linked immunosorbent assay; ICAM, intercellular adhesion molecule; IL, interleukin; IκB, inhibitor of κB; MOI, multiplicity of infection; NF-κB, nuclear factor-κB; PH, pleckstrin homology; PI, phosphatidylinositol; PI(4,5)P2, PI 4,5-bisphosphate; PI(3,4,5)P3, PI 3,4,5-trisphosphate; PBS, phosphate-buffered saline; TCID, tissue culture infectivity dose; TNF, tumor necrosis factor; IKK, IκB kinase. is a single-stranded RNA virus from the Picornaviridae family responsible for the majority of common colds. Viral infections trigger the majority of asthma exacerbations (1Johnston S.L. Pattemore P.K. Sanderson G. Smith S. Lampe F. Josephs L. Symington P. O'Toole S. Myint S.H. Tyrrell D.A. Holgate S.T. Br. Med. J. 1995; 310: 1225-1229Crossref PubMed Scopus (1669) Google Scholar, 2Nicholson K.G. Kent J. Ireland D.C. Br. Med. J. 1993; 307: 982-986Crossref PubMed Scopus (947) Google Scholar), and RV accounts for 60% of virus-induced exacerbations (1Johnston S.L. Pattemore P.K. Sanderson G. Smith S. Lampe F. Josephs L. Symington P. O'Toole S. Myint S.H. Tyrrell D.A. Holgate S.T. Br. Med. J. 1995; 310: 1225-1229Crossref PubMed Scopus (1669) Google Scholar). RV is also an important trigger of chronic obstructive pulmonary disease (COPD) exacerbations (3Seemungal T. Harper-Owen R. Bhowmik A. Moric I. Sanderson G. Message S. Maccallum P. Meade T.W. Jeffries D.J. Johnston S.L. Wedzicha J.A. Am. J. Respir. Crit. Care Med. 2001; 164: 1618-1623Crossref PubMed Scopus (855) Google Scholar, 4Rohde G. Wiethege A. Borg I. Kauth M. Bauer T.T. Gillissen A. Bufe A. Schultze-Werninghaus G. Thorax. 2003; 58: 37-42Crossref PubMed Scopus (460) Google Scholar). The precise mechanisms by which RV induces asthma or COPD exacerbations are unknown, but numerous studies suggest a role for IL-8, a CXC chemokine with the neutrophil-attractant Glu-Leu-Arg (ELR) motif. IL-8 and neutrophils are found in the nasal secretions and sputum of patients with RV-induced asthma exacerbations (5Grunberg K. Smits H.H. Timmers M.C. De Klerk E.P.A. Dolhain R.J.E.M. Dick E.C. Hiemstra P.S. Sterk P.J. Am. J. Respir. Crit. Care Med. 1997; 156: 609-616Crossref PubMed Scopus (166) Google Scholar, 6Grunberg K. Timmers M.C. Smits H.H. de Klerk E.P. Dick E.C. Spaan W.J. Hiemstra P.S. Sterk P.J. Clin. Exp. Allergy. 1997; 27: 36-45Crossref PubMed Scopus (170) Google Scholar, 7Pizzichini M.M. Pizzichini E. Efthimiadis A. Chauhan A.J. Johnston S.L. Hussack P. Mahony J. Dolovich J. Hargreave F.E. Am. J. Respir. Crit. Care Med. 1998; 158: 1178-1184Crossref PubMed Scopus (185) Google Scholar, 8Fleming H. Little F. Schnurr D. Avila P. Wong H. Liu J. Yagi S. Boushey H. Am. J. Respir. Crit. Care Med. 1999; 160: 100-108Crossref PubMed Scopus (113) Google Scholar, 9Gern J.E. Vrtis R. Grindle K.A. Swenson C. Busse W.W. Am. J. Respir. Crit. Care Med. 2000; 162: 2226-2231Crossref PubMed Scopus (239) Google Scholar). Further, the number of neutrophils correlates with the level of IL-8 (7Pizzichini M.M. Pizzichini E. Efthimiadis A. Chauhan A.J. Johnston S.L. Hussack P. Mahony J. Dolovich J. Hargreave F.E. Am. J. Respir. Crit. Care Med. 1998; 158: 1178-1184Crossref PubMed Scopus (185) Google Scholar, 9Gern J.E. Vrtis R. Grindle K.A. Swenson C. Busse W.W. Am. J. Respir. Crit. Care Med. 2000; 162: 2226-2231Crossref PubMed Scopus (239) Google Scholar). RV induces IL-8 expression in cultured airway epithelial cells (10Subauste M.C. Jacoby D.B. Richards S.M. Proud D. J. Clin. Invest. 1995; 96: 549-557Crossref PubMed Scopus (353) Google Scholar, 11Zhu Z. Tang W. Gwaltney J.M. Wu Y. Elias J.A. Am. J. Physiol. 1997; 273 (-L824): L814PubMed Google Scholar, 12Yamaya M. Sekizawa K. Suzuki T. Yamada N. Furukawa M. Ishizuka S. Nakayama K. Terajima M. Numazaki Y. Sasaki H. Am. J. Physiol. 1999; 277 (-L371): L362PubMed Google Scholar). Increased neutrophil and IL-8 levels are a feature of asthma (13Norzila M.Z. Fakes K. Henry R.L. Simpson J. Gibson P.G. Am. J. Respir. Crit. Care Med. 2000; 161: 769-774Crossref PubMed Scopus (204) Google Scholar, 14Ordonez C.L. Shaughnessy T.E. Matthay M.A. Fahy J.V. Am. J. Respir. Crit. Care Med. 2000; 161: 1185-1190Crossref PubMed Scopus (370) Google Scholar) and COPD exacerbations (15Gompertz S. O'Brien C. Bayley D.L. Hill S.L. Stockley R.A. Eur. Respir. J. 2001; 17: 1112-1119Crossref PubMed Scopus (264) Google Scholar, 16Aaron S.D. Angel J.B. Lunau M. Wright K. Fex C. Le Saux N. Dales R.E. Am. J. Respir. Crit. Care Med. 2001; 163: 349-355Crossref PubMed Scopus (405) Google Scholar, 17Qiu Y. Zhu J. Bandi V. Atmar R.L. Hattotuwa K. Guntupalli K.K. Jeffery P.K. Am. J. Respir. Crit. Care Med. 2003; 168: 968-975Crossref PubMed Scopus (299) Google Scholar). Together, these data suggest that RV may stimulate asthma exacerbations by inducing bronchial epithelial cell production of IL-8, leading to a neutrophilic inflammatory response. The human RVs include more than 100 serotypes that are divided into two groups based on their cellular receptors. Intercellular adhesion molecule (ICAM)-1 is the airway epithelial cell receptor for major subgroup RVs (e.g. RV14, RV16, and RV39), whereas the low density lipoprotein receptor functions as the receptor for minor subgroup RVs (e.g. RV1B and RV2). Thus, antibodies to ICAM-1 inhibit major subgroup RV infection of epithelial cells (18Terajima M. Yamaya M. Sekizawa K. Okinaga S. Suzuki T. Yamada N. Nakayama K. Ohrui T. Oshima T. Numazaki Y. Sasaki H. Am. J. Physiol. 1997; 273 (-L759): L749PubMed Google Scholar). ICAM-1 is a type I transmembrane glycoprotein belonging to the immunoglobulin superfamily. In endothelial cells, ICAM-1 cross-linking increases phosphorylation and activation of pp60 Src (19Wang Q. Pfeiffer G.R. Gaarde W.A. J. Biol. Chem. 2003; 278: 47731-47743Abstract PubMed Scopus Google Scholar, S. P. J. J. 2000; PubMed Scopus Google Scholar) and adhesion S. P. J. S. J. 1998; 161: Google Scholar). The of PI 3-kinase as a for Src R. M.C. J. Biol. Chem. 1995; PubMed Scopus Google Scholar) and adhesion S. A. PubMed Scopus Google Scholar, P. H. J. Biol. Chem. PubMed Scopus Google Scholar), suggesting that ICAM-1 by RV also PI 3-kinase. with also PI 3-kinase. and the and phosphorylation of adhesion and the of PI as well as activation of the PI 3-kinase in human cells E. D. R. D.A. G.R. J. 1998; PubMed Google Scholar). of which to PI 3-kinase in cells S. P.K. J. T. J. 2000; PubMed Scopus Google Scholar). virus PI 3-kinase in cells M.M. J.M. T. J. Biol. Chem. PubMed Scopus Google Scholar). these data suggest that RV to ICAM-1 IL-8 expression in by activation of PI PI a domain that with PI, and the PI 3-kinase inhibitor of the PI 3-kinase family are divided into to their and in as in M. M. 2003; PubMed Scopus Google PI and are with a or that Src homology with for in found in their and The major for the I PI in to PI(4,5)P2, and the major to C.L. PubMed Scopus Google Scholar). PI cellular functions by to the of these the of which is the Akt, to 3-phosphorylated a pleckstrin homology We examined the of PI 3-kinase and Akt to RV-induced responses in human airway epithelial We found that infection with RV39 induced rapid activation of PI 3-kinase and phosphorylation of Akt, as well as of 3-phosphorylated PI at the site of RV infection. Inhibition of PI 3-kinase and Akt attenuated RV39-induced nuclear transactivation and IL-8 expression. Inhibition of PI 3-kinase also blocked internalization of labeled suggesting that the requirement of PI 3-kinase for RV39-induced IL-8 expression, at least in part, relates to its role in viral endocytosis. human bronchial epithelial cells from bronchial epithelial tissue with the A. M. K. Ohrui T. L. K. W. J. D. Am. J. Respir. Biol. PubMed Scopus Google Scholar) by of The cells in with and of human bronchial epithelial cell from cultured at an as S. J. PubMed Scopus Google Scholar). cells into The cells in bronchial epithelial cell and the cells to a of bronchial epithelial cell and the and and at to The is with cells the of cells and and with to that the is also an of RV from Viral by cells with RV in of the cells cell and cellular by for at of cell RV and by with a for at A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar). virus to the and is PI 3-kinase activation or phosphorylation of Akt, cell than to the of by cellular fluorescent and RV39 on a of and and for at The and the in of phosphate-buffered divided into and at by with RV to and infection. The cells with RV39 for on at 16HBE14o- cells with RV39 at a multiplicity of infection of at a tissue culture infectivity of to by the S.L. Tyrrell for and Scholar). the cells with an of cell or In RV39 on for a at A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar). The of by the of a on cell for with the PI 3-kinase inhibitor in from or In the cells with for to or tumor necrosis infection. a of of IL-8 cells to for and with RV39 for with to cell and at RV to the for and from the IL-8 protein levels by enzyme-linked immunosorbent of IL-8 and NF-κB of the human IL-8 into from M.A. 1998; PubMed Google Scholar). NF-κB from family from a PI and an PI 3-kinase with a from which its S. J. Biol. 1999; PubMed Scopus Google Scholar), from which a and which an in which and to by by of J. S.L. C. D.L. Biol. 2003; PubMed Scopus Google Scholar). of inhibitor of encoding and by J.A. D.C. S.M. Biol. 1995; PubMed Google Scholar). IκB in which and by S. J.A. A. Biol. 1998; PubMed Google Scholar), from 16HBE14o- cells to in and with and the with with The the cells for The cells with RV39 for with and the cells for analysis a in for by by The as the of by not RNA from human epithelial cell on RNA and from RNA from cells by with the IL-8 and with as by the PI RV 16HBE14o- cells with with and for at The with of and of The by at for in of and to The and by the 16HBE14o- cells cellular by and to a The with antibodies Akt, or PI 3-kinase The and with and fluorescent of of to a of virus to to The virus in the for with of an The with of and labeled virus the at The virus by with a in for at and at for at The and the virus in of phosphate-buffered with for virus with in with RV39 internalization into 16HBE14o- cells, cells on with virus or an of cell from cells for at The cells in with for at with PBS, and with and by fluorescent We also the infection of 16HBE14o- cells with fluorescent protein or V. T. T. T. S. J. Biol. 2001; PubMed Scopus Google Scholar, J.A. Biol. PubMed Scopus Google Scholar), a citrogen-tagged fusion protein encoding the pleckstrin homology domain of Akt by of infection with labeled the cells and as and by fluorescent by from and with and from 16HBE14o- cells, and as J. S. A. A. Am. J. Physiol. 2003; PubMed Scopus Google Scholar). nuclear at for in of and the to for on a and at for in The and by from cell with an of the in of in the of of and as a for as S. J.A. A. Biol. 1998; PubMed Google Scholar, J. S. A. A. Am. J. Physiol. 2003; PubMed Scopus Google Scholar). The with by and the to a by the to and phosphorylation by levels of by of RV39 and virus by cells with RV39 in the of and for the cells by and by with a as 16HBE14o- cells RV39 for at or at G. M. L. D. R. J. PubMed Scopus Google Scholar). virus 16HBE14o- cells in with in of for at The cells at with PBS, and in cell by 16HBE14o- cells in with and with cells in with at for The cells to for by with to and for of by analysis of by by the RV39 Infection IL-8 in human bronchial epithelial cells with RV39 at cell for of IL-8 protein by RV39 IL-8 in a of cell from cells of RV39 as low as IL-8 protein not IL-8 expression cells with the of the human IL-8 into a with RV39 at and for an RV39 IL-8 by with the that IL-8 is by RV39 infection. We RV39 infection increases IL-8 expression in human epithelial cells, from the of at for RV39 from the and from the and IL-8 by RV39 and IL-8 protein with IL-8 IL-8 level also in with RV39 that the increases in IL-8 expression to RV39 the cells with to RV39 infection. of used as a with but not attenuated IL-8 expression RV39-induced IL-8 expression from the expression of cells with of on IL-8 expression the IL-8 expression in a IL-8 protein expression and infection with of attenuated but not IL-8 expression and activation and the viral to of IL-8 protein expression to that by suggesting that viral not for the IL-8 response. PI and Akt by RV39 RV39 16HBE14o- cells for with with and with and the to the role of the of PI also for a cells with RV infection PI phosphorylation and phosphorylation of of infection We also examined RV39 infection induces phosphorylation of 16HBE14o- cellular protein to to and with an RV39 induced Akt which not to a in Akt expression, with the that RV39 of RV39 as low as Akt RV39-induced than from the of PI 3-kinase and Akt, used RV39 to 16HBE14o- cells with fluorescent protein or a fluorescent fusion protein encoding the pleckstrin homology domain of In cells, virus colocalized with as by the and cells to of 100 cells with cell 16HBE14o- cells with fluorescent protein and with labeled RV39 Together, these data that 3-phosphorylated PI accumulates at the site of RV infection and that RV infection PI 3-kinase and PI and Akt for RV39-induced IL-8 examined the requirement of PI 3-kinase activation for RV39-induced responses by cells with a inhibitor of PI that Akt phosphorylation by in the of RV Akt functions downstream of PI 3-kinase In 16HBE14o- cells, for attenuated RV39-induced IL-8 protein expression in a also blocked IL-8 protein expression in of PI as well as the PI 3-kinase the role of I PI 16HBE14o- cells with and a encoding a of the of PI 3-kinase. of RV39-induced from the IL-8 We also examined the requirement of Akt for RV39-induced with and a of of RV39-induced from the IL-8 with the that RV39-induced IL-8 expression activation of a PI NF-κB for RV39-induced IL-8 human bronchial epithelial cells with RV39 and nuclear for RV39 the of nuclear to an encoding the NF-κB that RV39 induces NF-κB and We 16HBE14o- cells with a encoding into Infection with RV39 NF-κB transactivation the level of transactivation than that induced by We also 16HBE14o- cells with in which the NF-κB site of the NF-κB site to RV39 that RV39-induced IL-8 expression NF-κB We phosphorylation of RV infection. RV39 infection phosphorylation protein with by the of a of the phosphorylation but not a RV39-induced IL-8 that phosphorylation of is for the response. expression of a RV39-induced IL-8 these that RV39 may expression and that NF-κB signaling is for RV39-induced IL-8 expression. PI for NF-κB the requirement of PI 3-kinase for RV39-induced NF-κB 16HBE14o- cells also with the NF-κB and with or with a of the of PI 3-kinase and NF-κB transactivation and suggesting that PI 3-kinase is for the response. In with RV39-induced phosphorylation by the expression of PI 3-kinase and Akt for NF-κB transactivation and of PI for RV39 the of on of RV39 and In cells with a in labeled RV39 fluorescent as well with suggesting that PI 3-kinase is for viral We the of on and internalization of virus by 16HBE14o- cells with RV39 and at and of on RV39 with RV39 as by the of labeled virus to these data suggest that PI 3-kinase is for viral Viral infections trigger the majority of asthma exacerbations (1Johnston S.L. Pattemore P.K. Sanderson G. Smith S. Lampe F. Josephs L. Symington P. O'Toole S. Myint S.H. Tyrrell D.A. Holgate S.T. Br. Med. J. 1995; 310: 1225-1229Crossref PubMed Scopus (1669) Google Scholar, 2Nicholson K.G. Kent J. Ireland D.C. Br. Med. J. 1993; 307: 982-986Crossref PubMed Scopus (947) Google Scholar), and RV accounts for 60% of virus-induced exacerbations (1Johnston S.L. Pattemore P.K. Sanderson G. Smith S. Lampe F. Josephs L. Symington P. O'Toole S. Myint S.H. Tyrrell D.A. Holgate S.T. Br. Med. J. 1995; 310: 1225-1229Crossref PubMed Scopus (1669) Google Scholar). RV is also an important trigger of COPD exacerbations (3Seemungal T. Harper-Owen R. Bhowmik A. Moric I. Sanderson G. Message S. Maccallum P. Meade T.W. Jeffries D.J. Johnston S.L. Wedzicha J.A. Am. J. Respir. Crit. Care Med. 2001; 164: 1618-1623Crossref PubMed Scopus (855) Google Scholar, 4Rohde G. Wiethege A. Borg I. Kauth M. Bauer T.T. Gillissen A. Bufe A. Schultze-Werninghaus G. Thorax. 2003; 58: 37-42Crossref PubMed Scopus (460) Google Scholar). studies suggest that RV may stimulate exacerbations of asthma and COPD by inducing bronchial epithelial cell production of IL-8, leading to a neutrophilic inflammatory (5Grunberg K. Smits H.H. Timmers M.C. De Klerk E.P.A. Dolhain R.J.E.M. Dick E.C. Hiemstra P.S. Sterk P.J. Am. J. Respir. Crit. Care Med. 1997; 156: 609-616Crossref PubMed Scopus (166) Google Scholar, 6Grunberg K. Timmers M.C. Smits H.H. de Klerk E.P. Dick E.C. Spaan W.J. Hiemstra P.S. Sterk P.J. Clin. Exp. Allergy. 1997; 27: 36-45Crossref PubMed Scopus (170) Google Scholar, 7Pizzichini M.M. Pizzichini E. Efthimiadis A. Chauhan A.J. Johnston S.L. Hussack P. Mahony J. Dolovich J. Hargreave F.E. Am. J. Respir. Crit. Care Med. 1998; 158: 1178-1184Crossref PubMed Scopus (185) Google Scholar, 8Fleming H. Little F. Schnurr D. Avila P. Wong H. Liu J. Yagi S. Boushey H. Am. J. Respir. Crit. Care Med. 1999; 160: 100-108Crossref PubMed Scopus (113) Google Scholar, 9Gern J.E. Vrtis R. Grindle K.A. Swenson C. Busse W.W. Am. J. Respir. Crit. Care Med. 2000; 162: 2226-2231Crossref PubMed Scopus (239) Google Scholar, M.C. Jacoby D.B. Richards S.M. Proud D. J. Clin. Invest. 1995; 96: 549-557Crossref PubMed Scopus (353) Google Scholar, 11Zhu Z. Tang W. Gwaltney J.M. Wu Y. Elias J.A. Am. J. Physiol. 1997; 273 (-L824): L814PubMed Google Scholar, 12Yamaya M. Sekizawa K. Suzuki T. Yamada N. Furukawa M. Ishizuka S. Nakayama K. Terajima M. Numazaki Y. Sasaki H. Am. J. Physiol. 1999; 277 (-L371): L362PubMed Google Scholar, M.Z. Fakes K. Henry R.L. Simpson J. Gibson P.G. Am. J. Respir. Crit. Care Med. 2000; 161: 769-774Crossref PubMed Scopus (204) Google Scholar, 14Ordonez C.L. Shaughnessy T.E. Matthay M.A. Fahy J.V. Am. J. Respir. Crit. Care Med. 2000; 161: 1185-1190Crossref PubMed Scopus (370) Google Scholar, S. O'Brien C. Bayley D.L. Hill S.L. Stockley R.A. Eur. Respir. J. 2001; 17: 1112-1119Crossref PubMed Scopus (264) Google Scholar, 16Aaron S.D. Angel J.B. Lunau M. Wright K. Fex C. Le Saux N. Dales R.E. Am. J. Respir. Crit. Care Med. 2001; 163: 349-355Crossref PubMed Scopus (405) Google Scholar, 17Qiu Y. Zhu J. Bandi V. Atmar R.L. Hattotuwa K. Guntupalli K.K. Jeffery P.K. Am. J. Respir. Crit. Care Med. 2003; 168: 968-975Crossref PubMed Scopus (299) Google Scholar). Little is the signaling regulating RV-induced IL-8 expression, The IL-8 to infection in airway epithelial cells is by antibodies to that a role (18Terajima M. Yamaya M. Sekizawa K. Okinaga S. Suzuki T. Yamada N. Nakayama K. Ohrui T. Oshima T. Numazaki Y. Sasaki H. Am. J. Physiol. 1997; 273 (-L759): L749PubMed Google Scholar). NF-κB activation been to for a number of RV responses airway epithelial cell expression of IL-8, and cell adhesion Z. Tang W. Gwaltney J.M. Wu Y. Elias J.A. Am. J. Physiol. 1997; 273 (-L824): L814PubMed Google Scholar, A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar, Z. Tang W. A. W. J. Clin. Invest. PubMed Scopus Google Scholar, A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar, J. V. Proud D. J. 2000; PubMed Scopus Google Scholar). with a inhibitor of protein IL-8 expression in epithelial cells S.D. R. J. 2000; PubMed Scopus Google Scholar). The inhibitor of IL-8 J.E. D.A. Grindle K.A. R.A. Busse W.W. Am. J. Respir. Biol. 2003; PubMed Scopus Google Scholar). in that is to IL-8 NF-κB activation J.A. A. J. L. J. PubMed Scopus Google Scholar). In the have that RV39 infection induces activation of the PI signaling Infection of 16HBE14o- human bronchial epithelial cells with RV39 induced rapid activation of PI 3-kinase and phosphorylation of Akt, a downstream effector of PI 3-kinase. PI 3-kinase been to by in E. D. R. D.A. G.R. J. 1998; PubMed Google Scholar), virus S. P.K. J. T. J. 2000; PubMed Scopus Google Scholar), and virus M.M. J.M. T. J. Biol. Chem. PubMed Scopus Google Scholar), by of RV with cit-Akt-PH, that 3-phosphorylated PI accumulates at the site of viral infection. Further, in 16HBE14o- and airway epithelial cells that of PI by or expression of a of the of PI RV39-induced expression of Further, have that the requirement of PI 3-kinase for RV39-induced NF-κB transactivation and IL-8 expression, at least in part, relates to its role in viral with internalization of and PI 3-kinase been to for internalization of E. D. R. D.A. G.R. J. 1998; PubMed Google Scholar). of viral internalization to the of downstream as have NF-κB activation and IL-8 expression. PI than a may also downstream signaling leading to IL-8 expression. with activation of PI 3-kinase and Akt, a downstream effector of PI for NF-κB NF-κB been to for RV-induced expression of IL-8 and Z. Tang W. Gwaltney J.M. Wu Y. Elias J.A. Am. J. Physiol. 1997; 273 (-L824): L814PubMed Google Scholar, A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar, Z. Tang W. A. W. J. Clin. Invest. PubMed Scopus Google Scholar, A. Johnston S.L. J. Biol. Chem. 1999; PubMed Scopus Google Scholar, J. V. Proud D. J. 2000; PubMed Scopus Google Scholar). We have these by that phosphorylation of and are for RV39-induced IL-8 We have also found that expression of a NF-κB suggesting that is the of in Y. and M. We found in human epithelial cells that infection with RV39 induces a in IL-8 expression. data with a by and N. G. R. L. H. Yagi S. Schnurr D. Boushey Am. J. Physiol. PubMed Scopus Google Scholar) that cells are to RV infection and that RV to IL-8 the of RVs to human airway epithelial cells in the of cells to RV is The for may to the of the RV airway epithelial cells more the than as infection. to studies Am. J. Physiol. PubMed Google Scholar), an and expression of not We not the of RV-induced PI 3-kinase cross-linking of ICAM-1 in the phosphorylation of Src and adhesion (19Wang Q. Pfeiffer G.R. Gaarde W.A. J. Biol. Chem. 2003; 278: 47731-47743Abstract PubMed Scopus Google Scholar, S. P. J. J. 2000; PubMed Scopus Google Scholar, S. P. J. S. J. 1998; 161: Google Scholar), of which PI 3-kinase R. M.C. J. Biol. Chem. 1995; PubMed Scopus Google Scholar, S. A. PubMed Scopus Google Scholar, P. H. J. Biol. Chem. PubMed Scopus Google Scholar). not of the PI 3-kinase in the RV-induced response. of the PI 3-kinase IL-8 the of in the of from the IL-8 Further, is the major of the I PI in C.L. PubMed Scopus Google Scholar) and the 3-phosphorylated of the Akt the of RV39 and PI 3-kinase is that PI 3-kinase also RV which is on D. H. Med. 1997; PubMed Scopus Google Scholar, M. M. N. D. R. 2001; PubMed Scopus Google Scholar, T. Yamaya M. Sekizawa K. M. Yamada N. Ishizuka S. Nakayama K. M. Numazaki Y. Sasaki H. Am. J. Physiol. 2001; PubMed Google Scholar). PI 3-kinase which PI and PI in D.A. R.E. 1998; PubMed Scopus Google Scholar), is to and may I. Smith J. 2001; PubMed Scopus Google Scholar). The PI 3-kinase which PI as a and is responsible for the of PI is also for J.M. J. Biol. 2001; PubMed Scopus Google Scholar). Thus, of viral by in to of these PI 3-kinase on the that PI 3-kinase and Akt are that of ICAM-1 and may and viral may for at least a of responses to with RV for IL-8 expression, as been T. Yamaya M. Sekizawa K. M. Yamada N. Ishizuka S. Nakayama K. M. Numazaki Y. Sasaki H. Am. J. Physiol. 2001; PubMed Google Scholar). may RV airway in the of viral Gwaltney J.M. Am. Respir. Google Scholar). activation of PI 3-kinase signaling the to in the leading to or the of IL-8 and in to RV infection to in S.D. R. J. 2000; PubMed Scopus Google Scholar), is that the signaling on viral and are responsible for the to RV, to by a of We that PI 3-kinase a role in RV-induced airway epithelial cell PI 3-kinase a for in the of chronic obstructive pulmonary and chronic that are by RV infection. We and for the of and for We also for of