Lentiviral-mediated GATA-3 RNAi Decreases Allergic Airway Inflammation and Hyperresponsiveness

Abstract

GATA-3 is the key transcriptional factor for Th2 commitment in T cells and is strongly associated with asthma and allergic disease. We studied the silencing of the GATA-3 gene expression using RNA interference (RNAi) delivered by a lentiviral vector, to evaluate the therapeutic role of GATA-3 short hairpin RNAs (shRNAs) in a murine model of asthma. Mice were sensitized with OVA and instilled intratracheally (IT) with GATA-3 shRNAs lentiviral vector (Lenti-si-GATA-3) once, 48 hours before challenge. After three challenges with the OVA antigen, the mice were assessed for airway hyperresponsiveness (AHR) and inflammation. With infection of Lenti-si-GATA-3 in EL-4 cells, GATA-3 gene expression was abrogated and downstream Th2 cytokines, such as interleukin-4 (IL-4) and IL-5, were also significantly inhibited. IT delivery of Lenti-si-GATA-3 in OVA-immunized mice resulted in a strong inhibition of local GATA-3 gene expression. Treatment with Lenti-si-GATA-3 successfully alleviated OVA-induced airway eosinophilia and Th2 cytokine release. While evaluating AHR by means of enhanced pause (Penh) and pulmonary resistance (RL) using body plethysmography, it was found that the administration of Lenti-si-GATA-3 had significantly decreased AHR in OVA-immunized mice. These results suggest that inhibition of GATA-3 gene expression by shRNAs lentiviral vectors strongly attenuates antigen-induced airway inflammation and hyper-responsiveness in mice. GATA-3 is the key transcriptional factor for Th2 commitment in T cells and is strongly associated with asthma and allergic disease. We studied the silencing of the GATA-3 gene expression using RNA interference (RNAi) delivered by a lentiviral vector, to evaluate the therapeutic role of GATA-3 short hairpin RNAs (shRNAs) in a murine model of asthma. Mice were sensitized with OVA and instilled intratracheally (IT) with GATA-3 shRNAs lentiviral vector (Lenti-si-GATA-3) once, 48 hours before challenge. After three challenges with the OVA antigen, the mice were assessed for airway hyperresponsiveness (AHR) and inflammation. With infection of Lenti-si-GATA-3 in EL-4 cells, GATA-3 gene expression was abrogated and downstream Th2 cytokines, such as interleukin-4 (IL-4) and IL-5, were also significantly inhibited. IT delivery of Lenti-si-GATA-3 in OVA-immunized mice resulted in a strong inhibition of local GATA-3 gene expression. Treatment with Lenti-si-GATA-3 successfully alleviated OVA-induced airway eosinophilia and Th2 cytokine release. While evaluating AHR by means of enhanced pause (Penh) and pulmonary resistance (RL) using body plethysmography, it was found that the administration of Lenti-si-GATA-3 had significantly decreased AHR in OVA-immunized mice. These results suggest that inhibition of GATA-3 gene expression by shRNAs lentiviral vectors strongly attenuates antigen-induced airway inflammation and hyper-responsiveness in mice. IntroductionAsthma is a chronic inflammatory disease of the airway, with a prevalence and severity that have undergone a dramatic increase in both developing and developed countries.1Bousquet J Ndiaye M Aït-Khaled N Annesi-Maesano I Vignola AM Management of chronic respiratory and allergic diseases in developing countries. Focus on sub-Saharan Africa.Allergy. 2003; 58: 265-283Crossref PubMed Scopus (45) Google Scholar,2Masoli M Fabian D Holt S Beasley R Global Initiative for Asthma (GINA) Program. The global burden of asthma: executive summary of the GINA Dissemination Committee Report.Allergy. 2004; 59: 469-478Crossref PubMed Scopus (2505) Google Scholar Allergic asthma has been characterized by pulmonary inflammation, reversible airway obstruction, and airway hyperresponsiveness (AHR) to both specific and non-specific stimuli.3Bentley AM Menz G Storz C Robinson DS Bradley B Jeffery PK et al.Identification of Tlymphocytes, macrophages, and activated eosinophils in the bronchial mucosa in intrinsic asthma. Relationship to symptoms and bronchial responsiveness.Am Rev Respir Dis. 1992; 146: 500-506Crossref PubMed Scopus (383) Google Scholar,4Björnsdottir US Cypcar DM Asthma: an inflammatory mediator soup.Allergy. 1999; 54: 55-61Crossref PubMed Google Scholar,5Boushey HA Fahy JV Targeting cytokines in asthma therapy: round one.Lancet. 2000; 356: 2114-2116Abstract Full Text Full Text PDF PubMed Scopus (36) Google Scholar,6Eum SY Haile S Lefort J Huerre M Vargaftig BB Eosinophil recruitment into the respiratory epithelium following antigenic challenge in hyper-IgE mice is accompanied by interleukin 5-dependent bronchial hyperresponsiveness.Proc Natl Acad Sci USA. 1995; 92: 12290-12294Crossref PubMed Scopus (181) Google Scholar,7Wills-Karp M Immunologic basis of antigen-induced airway hyperresponsiveness.Annu Rev Immunol. 1999; 17: 255-281Crossref PubMed Scopus (953) Google Scholar Infiltration and activation of eosinophils, into the bronchial wall and lumen, have been linked to events considered to be central to its pathophysiology. In contrast to the effector function of eosinophils, both clinical and experimental studies of allergic inflammation/asthma suggest that CD4+ T helper (Th) 2 cells may be the major effector cells in airway inflammation that are associated with lung dysfunction, through the recruitment and activation of eosinophils.8Watanabe A Mishima H Renzi PM Xu LJ Hamid Q Martin JG Transfer of allergic airway responses with antigen-primed CD4+ but not CD8+ T cells in brown Norway rats.J Clin Invest. 1995; 96: 1303-1310Crossref PubMed Scopus (117) Google Scholar,9Robinson DS Hamid Q Ying S Tsicopoulos A Barkans J Bentley AM et al.Predominant Th2-like bronchoalveolar T-lymphocyte population in atopic asthma.N Engl J Med. 1992; 326: 298-304Crossref PubMed Scopus (2551) Google Scholar,10Corrigan CJ Kay AB T cells and eosinophils in the pathogenesis of asthma.Immunol Today. 1992; 13: 501-507Abstract Full Text PDF PubMed Scopus (537) Google ScholarThe pathologic role of Th2 cells is mediated through the release of Th2 cytokines, such as interleukin-4 (IL-4), IL-5, and IL-13, and these three have distinct roles in the pathogenesis of asthma. IL-4 is the key cytokine involved in Th2 differentiation,11Kopf M Le Gros G Bachmann M Laemars MC Bluethmann H Köhler G Disruption of the murine IL-4 gene blocks Th2 cytokine responses.Nature. 1993; 362: 245-248Crossref PubMed Scopus (1091) Google Scholar,12Seder RA Paul WE Acquisition of lymphokine-producing phenotype by CD41 T cells.Annu Rev Immunol. 1994; 12: 635-673Crossref PubMed Scopus (1516) Google Scholar,13Morel PA Oriss TB Cross-regulation between Th1 and Th2 cells.Crit Rev Immunol. 1998; 18: 275-303Crossref PubMed Google Scholar whereas IL-13 has been associated with mucus production and AHR in a murine model of asthma.14Grunig G Warnock M Wakil AE Venkayya R Brombacher F Rennick DM et al.Requirement for IL-13 independently of IL-4 in experimental asthma.Science. 1998; 282: 2261-2263Crossref PubMed Scopus (1719) Google Scholar,15Wills-Karp M Luyimbazi J Xu X Schofield B Neben TY Karp CL et al.Interleukin-13: central mediator of allergic asthma.Science. 1998; 282: 2258-2261Crossref PubMed Scopus (2378) Google Scholar Further, IL-5 is the most important cytokine in the regulation of proliferation, differentiation, and activation of eosinophils.16Coffman RL Seymour BW Hudak S Jackson J Rennick D Antibody to interleukin-5 inhibits helminth-induced eosinophilia in mice.Science. 1989; 245: 308-310Crossref PubMed Scopus (539) Google Scholar,17Dent LA Strath M Mellor AL Sanderson CJ Eosinophilia in transgenic mice expressing interleukin 5.J Exp Med. 1990; 172: 1425-1431Crossref PubMed Scopus (503) Google Scholar,18Foster PS Hogan SP Ramsay AJ Matthaei KI Young IG Interleukin-5 deficiency abolishes airways eosinophilia and lung damage in mouse asthma model.J Exp Med. 1996; 183: 195-201Crossref PubMed Scopus (1270) Google Scholar,19Kopf M Brombacher F Hodgkin PD Ramsay AJ Milbourne EA Dai WJ et al.IL-5 deficient mice have a development defect in CD5+B-1 cells and lack eosinophils but have normal antibody and cytotoxic T cell responses.Immunity. 1996; 4: 15-24Abstract Full Text Full Text PDF PubMed Scopus (531) Google Scholar IL-4, IL-5, and IL-13 genes are clustered together in both the mouse and human genomes, suggesting a coordinated regulation of these genes.20Rengarajan J Szabo SJ Glimcher LH Transcriptional regulation of Th1/Th2 polarization.Immunol Today. 2000; 21: 479-483Abstract Full Text Full Text PDF PubMed Scopus (363) Google ScholarThe gene expression of all three Th2 cytokines is regulated by the transcriptional factor GATA-3. Studies on human asthmatic patients have demonstrated a significant increase in the number of GATA-3 messenger RNA-positive cells in airways as compared to that in tissues from non-asthmatic individuals.21Nakamura Y Ghaffar O Olivenstein R Taha RA Soussi-Gounni A Zhang DH et al.Gene expression of the GATA-3 transcription factor is increased in atopic asthma.J Allergy Clin Immunol. 1999; 103: 215-222Abstract Full Text Full Text PDF PubMed Google Scholar In an animal model, antigen challenge increased the GATA-3 expression of pulmonary eosinophils.22Justice JP Borchers MT Lee JJ Y expression of IL-4, and IL-5 by eosinophils in the of allergic J 282: PubMed Scopus Google Scholar specific expression of a GATA-3 the and challenge airway DH R et of allergic inflammation in a murine model of asthma by expression of a of 1999; Full Text Full Text PDF PubMed Scopus Google Scholar and local with GATA-3 pulmonary inflammation and S HA M M et of allergic airway inflammation and hyperresponsiveness by local of GATA-3 Exp Med. PubMed Scopus Google therapeutic in allergic RNA (RNAi) of RNA to the of GATA-3 short hairpin RNA in a murine model of GATA-3 gene silencing and production of shRNAs in T cell by lentiviral studied the lentiviral shRNAs GATA-3 expression in the mouse cell EL-4 the EL-4 cells were with with to increase GATA-3 gene and with the of three of GATA-3 shRNAs (Lenti-si-GATA-3) in EL-4 cells, the GATA-3 RNA and expression significantly decreased compared with the and with the the lentiviral vector expressing the of and The of GATA-3 was compared to the and compared to the also GATA-3 silencing in the mouse lung cell in the of Lenti-si-GATA-3 cells inhibition of the GATA-3 RNA by and with of Th2 cytokines of lentiviral also the of Lenti-si-GATA-3 on Th2 cytokine release in EL-4 the cells were with three of the of IL-4 and IL-5 were significantly decreased compared with that of the and with with The of IL-4 were and compared to the and The of IL-5 and compared to the and lung GATA-3 RNA expression by of GATA-3 in the of Lenti-si-GATA-3 in a murine model of mice were sensitized to OVA by three and with OVA The mice were with an (IT) of of Lenti-si-GATA-3 lentiviral vector expressing the of 2 before the challenge the GATA-3 RNA expression was in the of GATA-3 expression in lung administration of of animal and challenge. RNA of lung tissues were and for GATA-3 RNA expression by The experimental of the and and the sensitized and with were as sensitized and with OVA an increase in GATA-3 RNA expression in compared to the and mice After with three of the GATA-3 RNA expression in significantly decreased and the of and compared to and all shRNAs the of expressing lentiviral vector in the in of airway inflammation by GATA-3 lentiviral the of GATA-3 in airway inflammation, the in bronchial 48 hours the of three OVA challenges on In and mice inflammatory cell was in and challenge with OVA the and in significantly increased A was found in and mice that the IT of Lenti-si-GATA-3 significantly decreased OVA-induced and the of and compared to the and In Lenti-si-GATA-3 also significantly decreased OVA-induced and the of was compared to the of bronchoalveolar and of lung in mice of GATA-3 short hairpin RNA cell were on and cell were assessed by were as were with and for of inflammatory cells the a of of the of lung that and a of inflammatory cell airways compared with the After with mice also an inhibition of inflammatory cell compared to the and delivery of GATA-3 decreased IL-5, and IL-13 in recruitment of eosinophils, and Th2 in the is an important of allergic PD R et a cytokine in a model of allergic airways Exp Med. 1994; PubMed Scopus Google an in cells and in interleukin Natl Acad Sci USA. 1995; 92: PubMed Scopus Google Scholar We the in that of Lenti-si-GATA-3 significantly and the of as compared to with the the We also assessed the of important Th2 cytokines, IL-5 and IL-13, in of of mice. of Lenti-si-GATA-3 significantly IL-5 and IL-13 and the of inhibition and compared with the In the production of in was not the in bronchoalveolar in were by were as the delivery of GATA-3 the development of Lenti-si-GATA-3 the development of AHR in a murine model of asthma. was assessed by body plethysmography, and pulmonary resistance assessed using body plethysmography, the challenge. mice sensitized and with OVA an increase in the enhanced pause (Penh) and lung resistance (RL) to compared to and mice hyperresponsiveness by GATA-3 short hairpin RNAs (shRNAs) lentiviral vectors in mice. hyperresponsiveness was by body were as the of the enhanced pause (Penh) in the of of three resistance as by body were as the of the pulmonary resistance (RL) in the of RL of three AHR was increased in and mice the lentiviral vector After with the of and RL were to of the the and AHR was and in of clinical mice were the hours the and AHR was assessed the challenge. The results that administration of Lenti-si-GATA-3 the challenge significantly the increase in compared with the and the to was in mice that the the challenge in mice that the before the challenge and specific and of gene studies have on using to evaluate its therapeutic in a of the of as therapeutic in in has and clinical and the has been by the and for local of for a PubMed Scopus Google et S HA M M et of allergic airway inflammation and hyperresponsiveness by local of GATA-3 Exp Med. PubMed Scopus Google Scholar successfully of GATA-3 in airway inflammation and hyperresponsiveness in mice. that of GATA-3 gene expression by as a therapeutic in allergic has been the most delivered from RNA results in specific of messenger using RNA J 2004; 4: PubMed Scopus Google Scholar in the therapeutic role of of GATA-3 in a murine model of in the lentiviral vector as the delivery in shRNAs for gene silencing T N RNA 13: PubMed Scopus Google H PM of vector for delivery of 2003; PubMed Scopus Google C J et to genes in cells, cells and transgenic mice by RNA 2003; PubMed Scopus Google Scholar In three of GATA-3 shRNAs delivered by lentiviral vectors (Lenti-si-GATA-3) in and in to therapeutic in allergic asthma. We found that all three GATA-3 gene expression not in in the EL-4 T cell but also in in lung a of three of Lenti-si-GATA-3 not a of in not delivery of of GATA-3 shRNAs was to a inhibition of the gene expression. also be a of the of in the three with of the of Lenti-si-GATA-3 for both in and in The that the of Lenti-si-GATA-3 in these the of GATA-3 gene expression in EL-4 cells and lung cytokine production in airway inflammation, and AHR the of in the was Lenti-si-GATA-3 successfully GATA-3 expression in EL-4 cells and and lung Th2 cytokine expression in cell and not the of be by studies that demonstrated that transcription such as and factor activated T with GATA-3 to Th2 cytokine gene J Szabo SJ Glimcher LH Transcriptional regulation of Th1/Th2 polarization.Immunol Today. 2000; 21: 479-483Abstract Full Text Full Text PDF PubMed Scopus (363) Google Sanderson CJ activation of human interleukin-5 on of an PubMed Scopus Google J O R The and of the 1999; PubMed Scopus Google D A G AM F et and IL-13 gene expression in cells by three of the transcription Immunol. 2003; PubMed Scopus Google Scholar these transcriptional not a role in Th2 gene as with to Th2 cytokines the was an increase in GATA-3 expression in EL-4 cells and a AHR for the be associated with activation of T in the In in the found a in the to the N H H Y H M et of Th2 activation in airway J PubMed Scopus Google Scholar an of IL-13 and inhibition of the airway that the of be of in We that the lentiviral vector in the that T as to increase the the studies found that lentiviral vectors shRNAs from and were to by expression of AJ S A AL R of an by vectors in 2003; PubMed Scopus Google S R of gene by 2004; PubMed Scopus Google Scholar Lenti-si-GATA-3 the assessed and A expression by means of in lung We found that the Lenti-si-GATA-3 2 increased in and gene expression but not compared to the studies have demonstrated that gene SP X and of gene expression by RNAs 2004; PubMed Scopus Google AL J J M et gene regulation by 2003; 21: PubMed Scopus Google Scholar not in is not of lentiviral vectors to as in the a with human with human is for the of human vectors in clinical involved in the disease has a major in Targeting messenger RNA the is a to is on the that messenger RNA of a The and delivery of as a for it to be as a clinical in the We demonstrated that the GATA-3 vector be a of allergic inflammation and AHR in a murine model of asthma. with a for developing gene silencing for asthma and allergic and and experimental mice were from the of the of The Committee of the the and The mice were sensitized by of of OVA in 2 of in a of with of OVA in 2 of and with OVA in a of by administration on GATA-3 lentiviral vectors were into the 2 before the challenge with A cell were as of was by not The of administration was by the of expression. After 2 of IT of into OVA-immunized the of expression was in cells by and expression in the and of and of shRNAs into lentiviral shRNAs were for GATA-3 and The of GATA-3 is is the from These shRNAs were by expression to the shRNAs were into The lentiviral vectors in the were were by of T Y LJ lentiviral vectors with and 1999; Scopus Google Scholar lentiviral vector production was by of cells were with of vector helper and The were and by The were on cells on expression. The of vectors for of EL-4 cells was the of infection and cells were the of infection was into and by using The a significant increase in GATA-3 was and to the for GATA-3 and were using a and by The were with antibody GATA-3 and of cytokine expression. IL-5, IL-13, and were by the to the and lung using instilled with a that was by three and Y R N H T et in airway inflammation by Respir 1998; PubMed Scopus Google Scholar cell were assessed on The cells were using and with A of cells were were for The were with and were and with and to the number of inflammatory cells body was in by body mice were in the and were and for in were through an of the for were and for of were to the respiratory in was as and were as the of compared to from three J A A et of airway in allergic mice using J Respir PubMed Scopus Google of airway resistance in mice. resistance was assessed as an increase in pulmonary resistance challenge with in mice using a of the by et T A H R et of pulmonary to challenge in 2004; PubMed Scopus Google Scholar Mice were with and a of with a of and a of with a animal model was into the to the of the with a was by of and were resistance was by a was with an and through the The resistance of the was from all airway resistance were as the pulmonary resistance (RL) in the of RL of three to using of The of was a with Lenti-si-GATA-3 airway hyperresponsiveness in mice were of lentiviral vectors OVA challenge. hyperresponsiveness was by body as of the in the of with inhibition of gene expression and cytokines release in EL-4 cells and asthma in were with and of GATA-3 and lentiviral After of EL-4 cells were with with for GATA-3 RNA was by and Th2 cytokines release was by were as vector expressing GATA-3 Mice were sensitized with OVA and instilled intratracheally of of lentiviral vector expressing GATA-3 and with to in were by cell were on and cell were assessed by hyperresponsiveness was by body were as with and RNA expression in Lenti-si-GATA-3 lung were of lentiviral vectors to the in RNA of lung tissues were and for GATA-3 RNA expression by to the with of cells in delivery of expression lentiviral cells were assessed for expression of and Eosinophil with with with with and IntroductionAsthma is a chronic inflammatory disease of the airway, with a prevalence and severity that have undergone a dramatic increase in both developing and developed countries.1Bousquet J Ndiaye M Aït-Khaled N Annesi-Maesano I Vignola AM Management of chronic respiratory and allergic diseases in developing countries. Focus on sub-Saharan Africa.Allergy. 2003; 58: 265-283Crossref PubMed Scopus (45) Google Scholar,2Masoli M Fabian D Holt S Beasley R Global Initiative for Asthma (GINA) Program. The global burden of asthma: executive summary of the GINA Dissemination Committee Report.Allergy. 2004; 59: 469-478Crossref PubMed Scopus (2505) Google Scholar Allergic asthma has been characterized by pulmonary inflammation, reversible airway obstruction, and airway hyperresponsiveness (AHR) to both specific and non-specific stimuli.3Bentley AM Menz G Storz C Robinson DS Bradley B Jeffery PK et al.Identification of Tlymphocytes, macrophages, and activated eosinophils in the bronchial mucosa in intrinsic asthma. Relationship to symptoms and bronchial responsiveness.Am Rev Respir Dis. 1992; 146: 500-506Crossref PubMed Scopus (383) Google Scholar,4Björnsdottir US Cypcar DM Asthma: an inflammatory mediator soup.Allergy. 1999; 54: 55-61Crossref PubMed Google Scholar,5Boushey HA Fahy JV Targeting cytokines in asthma therapy: round one.Lancet. 2000; 356: 2114-2116Abstract Full Text Full Text PDF PubMed Scopus (36) Google Scholar,6Eum SY Haile S Lefort J Huerre M Vargaftig BB Eosinophil recruitment into the respiratory epithelium following antigenic challenge in hyper-IgE mice is accompanied by interleukin 5-dependent bronchial hyperresponsiveness.Proc Natl Acad Sci USA. 1995; 92: 12290-12294Crossref PubMed Scopus (181) Google Scholar,7Wills-Karp M Immunologic basis of antigen-induced airway hyperresponsiveness.Annu Rev Immunol. 1999; 17: 255-281Crossref PubMed Scopus (953) Google Scholar Infiltration and activation of eosinophils, into the bronchial wall and lumen, have been linked to events considered to be central to its pathophysiology. In contrast to the effector function of eosinophils, both clinical and experimental studies of allergic inflammation/asthma suggest that CD4+ T helper (Th) 2 cells may be the major effector cells in airway inflammation that are associated with lung dysfunction, through the recruitment and activation of eosinophils.8Watanabe A Mishima H Renzi PM Xu LJ Hamid Q Martin JG Transfer of allergic airway responses with antigen-primed CD4+ but not CD8+ T cells in brown Norway rats.J Clin Invest. 1995; 96: 1303-1310Crossref PubMed Scopus (117) Google Scholar,9Robinson DS Hamid Q Ying S Tsicopoulos A Barkans J Bentley AM et al.Predominant Th2-like bronchoalveolar T-lymphocyte population in atopic asthma.N Engl J Med. 1992; 326: 298-304Crossref PubMed Scopus (2551) Google Scholar,10Corrigan CJ Kay AB T cells and eosinophils in the pathogenesis of asthma.Immunol Today. 1992; 13: 501-507Abstract Full Text PDF PubMed Scopus (537) Google ScholarThe pathologic role of Th2 cells is mediated through the release of Th2 cytokines, such as interleukin-4 (IL-4), IL-5, and IL-13, and these three have distinct roles in the pathogenesis of asthma. IL-4 is the key cytokine involved in Th2 differentiation,11Kopf M Le Gros G Bachmann M Laemars MC Bluethmann H Köhler G Disruption of the murine IL-4 gene blocks Th2 cytokine responses.Nature. 1993; 362: 245-248Crossref PubMed Scopus (1091) Google Scholar,12Seder RA Paul WE Acquisition of lymphokine-producing phenotype by CD41 T cells.Annu Rev Immunol. 1994; 12: 635-673Crossref PubMed Scopus (1516) Google Scholar,13Morel PA Oriss TB Cross-regulation between Th1 and Th2 cells.Crit Rev Immunol. 1998; 18: 275-303Crossref PubMed Google Scholar whereas IL-13 has been associated with mucus production and AHR in a murine model of asthma.14Grunig G Warnock M Wakil AE Venkayya R Brombacher F Rennick DM et al.Requirement for IL-13 independently of IL-4 in experimental asthma.Science. 1998; 282: 2261-2263Crossref PubMed Scopus (1719) Google Scholar,15Wills-Karp M Luyimbazi J Xu X Schofield B Neben TY Karp CL et al.Interleukin-13: central mediator of allergic asthma.Science. 1998; 282: 2258-2261Crossref PubMed Scopus (2378) Google Scholar Further, IL-5 is the most important cytokine in the regulation of proliferation, differentiation, and activation of eosinophils.16Coffman RL Seymour BW Hudak S Jackson J Rennick D Antibody to interleukin-5 inhibits helminth-induced eosinophilia in mice.Science. 1989; 245: 308-310Crossref PubMed Scopus (539) Google Scholar,17Dent LA Strath M Mellor AL Sanderson CJ Eosinophilia in transgenic mice expressing interleukin 5.J Exp Med. 1990; 172: 1425-1431Crossref PubMed Scopus (503) Google Scholar,18Foster PS Hogan SP Ramsay AJ Matthaei KI Young IG Interleukin-5 deficiency abolishes airways eosinophilia and lung damage in mouse asthma model.J Exp Med. 1996; 183: 195-201Crossref PubMed Scopus (1270) Google Scholar,19Kopf M Brombacher F Hodgkin PD Ramsay AJ Milbourne EA Dai WJ et al.IL-5 deficient mice have a development defect in CD5+B-1 cells and lack eosinophils but have normal antibody and cytotoxic T cell responses.Immunity. 1996; 4: 15-24Abstract Full Text Full Text PDF PubMed Scopus (531) Google Scholar IL-4, IL-5, and IL-13 genes are clustered together in both the mouse and human genomes, suggesting a coordinated regulation of these genes.20Rengarajan J Szabo SJ Glimcher LH Transcriptional regulation of Th1/Th2 polarization.Immunol Today. 2000; 21: 479-483Abstract Full Text Full Text PDF PubMed Scopus (363) Google ScholarThe gene expression of all three Th2 cytokines is regulated by the transcriptional factor GATA-3. Studies on human asthmatic patients have demonstrated a significant increase in the number of GATA-3 messenger RNA-positive cells in airways as compared to that in tissues from non-asthmatic individuals.21Nakamura Y Ghaffar O Olivenstein R Taha RA Soussi-Gounni A Zhang DH et al.Gene expression of the GATA-3 transcription factor is increased in atopic asthma.J Allergy Clin Immunol. 1999; 103: 215-222Abstract Full Text Full Text PDF PubMed Google Scholar In an animal model, antigen challenge increased the GATA-3 expression of pulmonary eosinophils.22Justice JP Borchers MT Lee JJ Y expression of IL-4, and IL-5 by eosinophils in the of allergic J 282: PubMed Scopus Google Scholar specific expression of a GATA-3 the and challenge airway DH R et of allergic inflammation in a murine model of asthma by expression of a of 1999; Full Text Full Text PDF PubMed Scopus Google Scholar and local with GATA-3 pulmonary inflammation and S HA M M et of allergic airway inflammation and hyperresponsiveness by local of GATA-3 Exp Med. PubMed Scopus Google therapeutic in allergic RNA (RNAi) of RNA to the of GATA-3 short hairpin RNA in a murine model of asthma.