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Prostaglandin I2 Receptor (IP) Signaling Inhibits Lung Type I Interferon Expression by RSV Infection
Abstracts AB191
J ALLERGY CLIN IMMUNOL VOLUME 131, NUMBER 2
Prostaglandin I2 Receptor (IP) Signaling Inhibits Lung Type I Interferon Expression by RSV Infection Shinji Toki, PhD1, Sara Reiss1, Kasia Goleniewska, MS1, Martin L. Moore, PhD2, Garret FitzGerald, MD3, R. Stokes Peebles, Jr, MD, FAAAAI1; 1Vanderbilt University School of Medicine, Nashville, TN, 2 Emory University, Atlanta, GA, 3University of Pennsylvania, Philadelphia, PA. RATIONALE: Our laboratory has previously shown that prostaglandin I2 (PGI2) – IP signaling reduced RSV-induced illness and inflammation in a mouse model. However, the mechanisms of this down-regulation are not yet fully understood. In addition, the effect of PGI2 signaling on type I interferon (IFN) expression has not been reported. We hypothesized that PGI2–IP signaling decreases RSV-induced type I IFN (a and b) induction. To test this hypothesis, we measured airway and lung IFN-a and b protein expression. METHODS: BALB/c background WT and IPKO mice were challenged intranasally with RSV A2001/2-20 strain. Bronchoalveolar lavage (BAL) fluid and lung were harvested after 12 and 24 hours. Control mice were not challenged with RSV. IFN-a and b protein expression in the BAL fluid and lung homogenate were measured by ELISA. RESULTS: IPKO mice had significantly greater BAL IFN-a and b protein expression compared to WT mice (p<0.05) 12 hours after intranasal challenge of RSV. Lung IFN-a protein expression was increased in IPKO mice compared to WT mice (p<0.05), but IFN-b was not significantly different. There was a trend for a difference of both BAL and lung IFN-a and b protein expression 24 hours after RSV challenge. CONCLUSIONS: IPKO mice had significantly increased IFN-a and b protein expression in the airway and lung. This result suggests that endogenous PGI2 signaling down-regulates RSV-induced illness through suppression of type I IFN induction.
682
Rhinovirus Induces Differential Expression of Th2-Promoting Epithelial Cytokines From Asthmatics Ex Vivo Joshua L. Kennedy, MD1, Larry Borish, MD, FAAAAI2, Peter W. Heymann, MD1, John W. Steinke, PhD, FAAAAI2; 1University of Virginia, Charlottesville, VA, 2Asthma and Allergic Disease Center, Carter Center for Immunology Research, University of Virginia, Charlottesville, VA. RATIONALE: The majority of childhood asthma exacerbations are provoked by rhinovirus (RV) infections, and these most often occur when allergies are also present. We propose this reflects the propensity of RV infection of epithelium to induce expression of Th2-promoting cytokines and that this occurs without altering the innate immune response of these cells to RV. METHODS: Epithelial cells derived from sinonasal epithelium of asthmatics and non-asthmatics were cultured and infected with RV39. Cells and supernatants were collected at various time intervals. Quantitative PCR was used to measure gene expression for the Th2-inducing cytokines IL-25, IL-33, and TSLP, and innate immune cytokines IL-15, IFN-ß, and IL-18. Supernatants were assayed for cytokine protein secretion. RESULTS: mRNA levels for IL-25, IL-33, and TSLP were increased in asthmatics compared to non-asthmatics after infection with RV39 at 2 (IL25), 4 (TSLP), and 24 hours (IL-33). mRNA levels were increased for IL15, IFN-ß, and IL-18 though no differences were observed between asthmatics and non-asthmatics. Protein expression of IL-15 was comparably evident in both cohorts at 48 hours. CONCLUSIONS: In contrast to non-asthmatics, RV-infected sinonasal epithelium derived from asthmatics express mRNA for cytokines important in the induction of Th2 immune deviation. Additionally, there was little difference in the expression of innate cytokines, specifically IL-15 and IL-18, which suggests that there is no defect in innate immune responses to RV amongst asthmatics. We propose instead that it is this RVinduced barrage of Th2-inducing cytokines that is responsible for asthma exacerbations.
683
Genetic Variants in Thymic Stromal Lymphopoietin (TSLP) and Receptor (TSLPR) and Their Influence On the Humoral Immune Response Luis Fang, Cesar Mu~noz, Luz Hernandez, Beatriz Martinez, Javier Marrugo; University of Cartagena, Cartagena, Colombia. RATIONALE: TSLP/TSLPR participates in the initiation of the Th2immune response. Recently, genetic variants of these genes have been associated with atopic diseases. However, it’s unknown whether these polymorphisms influence humoral immune response. This study evaluated the influence of polymorphisms in TSLP and TSLPR on the antibody response against dietary antigens, Blomia tropicalis and Ascaris lumbricoides. METHODS: By RT-qPCR and TaqManÒ probes, we genotyped the SNPs: rs1837253 [C/T], rs17551370 [A/G], rs2289276 [C/T] in TSLP and rs36139698 [A/G], rs36177645 [C/T], and rs36133495 [A/G] in TSLPR in 80 African-descent individuals. The serum levels of specific IgE, IgA and IgG4 against Blomia tropicalis, egg, milk, and peanut extracts and IgE against Ascaris lumbricoides extract, were measured by ELISA. Data analysis was performed using principal component analysis (PCA) by R Project software. RESULTS: The minor allele frequencies were as following: rs1837253 [T:0.41.9%], rs7551370 [A:14.4%], rs2289276 [T:22.5%], rs36139698 [A:37%], rs36177645 [C:24%] and rs36133495 [G:45%]. The PCA showed that specific IgE-IgG4 responses to dietary antigens were different from B. tropicalis response, while specific IgA response was similar in both groups of antigens, furthermore IgE response against B. tropicalis and A. lumbricoides were positively correlated. The genotype CT/TT in rs1837253 influenced IgE-IgG4 responses independent of antigenic group, while GG in rs17551370, CT/TT in rs2289276, AG/GG in rs36139698 and CT/TT in rs36177645 strongly favor IgE-IgG4 responses to egg and peanuts extracts. Only AG/GG in rs36133495 influenced IgA response against dietary and B. tropicalis antigens. CONCLUSIONS: TSLP/TSLPR variants influence the IgE-IgG4-IgA immune responses against dietary and environment antigens in African-descent individuals from Colombia.
684
Immune Modulatory Effects of IL-22 On Allergen-Induced Pulmonary Inflammation Ping Fang; Asthma & allergy center, Johns Hopkins University School of Medicine. RATIONALE: Allergen-induced pulmonary inflammation in asthma is mainly TH2 dominated. A recently discovered TH17/TH22 cell-derived cytokine, IL-22, has been found to be increased in asthma. However, several studies showed controversial findings in the immune modulatory effects of IL-22 in animal models of allergic asthma. Our objective was to determine the regulatory role of IL-22 in OVA-induced allergic inflammation using a novel lung-specific IL-22 transgenic overexpression system. METHODS: Inducible IL-22 transgenic (CC10-rtTA- and SPC-rtTA-TREtight-IL-22) mice on C57BL/6 genetic background were generated. IHC of lung tissue and ELISA of bronchoalveolar lavage (BAL) fluid for IL-22 were performed to confirm the location and quantity of IL-22 expression. OVAinduced pulmonary inflammatory responses were compared between IL-22 transgenic and wild type control mice. BAL total and differential cell counts, lung histology, mucous metaplasia, IHC of major basic protein for eosinophils, and serum OVA-specific IgE were determined. RESULTS: IL-22 transgene was successfully activated in the large (CC10 promoter) and small (SPC promoter) airway epithelium cells by doxycycline in the drinking water. OVA sensitization and challenge did not induce endogenous IL-22 expression. Compared to wild type mice, IL-22 transgenic mice showed decreased percentage of eosinophils in the BAL and slight reduction in eosinophilic inflammation and mucus metaplasia in the airways. Interestingly, inflammatory cell infiltration in lung parenchyma appeared slightly accentuated. In addition, there was no statistic difference in serum OVA-specific IgE. CONCLUSIONS: These findings indicate that IL-22 may have immune modulatory effects on pulmonary inflammatory responses in the development of allergen-induced asthma.
MONDAY
681
J ALLERGY CLIN IMMUNOL VOLUME 131, NUMBER 2
Prostaglandin I2 Receptor (IP) Signaling Inhibits Lung Type I Interferon Expression by RSV Infection Shinji Toki, PhD1, Sara Reiss1, Kasia Goleniewska, MS1, Martin L. Moore, PhD2, Garret FitzGerald, MD3, R. Stokes Peebles, Jr, MD, FAAAAI1; 1Vanderbilt University School of Medicine, Nashville, TN, 2 Emory University, Atlanta, GA, 3University of Pennsylvania, Philadelphia, PA. RATIONALE: Our laboratory has previously shown that prostaglandin I2 (PGI2) – IP signaling reduced RSV-induced illness and inflammation in a mouse model. However, the mechanisms of this down-regulation are not yet fully understood. In addition, the effect of PGI2 signaling on type I interferon (IFN) expression has not been reported. We hypothesized that PGI2–IP signaling decreases RSV-induced type I IFN (a and b) induction. To test this hypothesis, we measured airway and lung IFN-a and b protein expression. METHODS: BALB/c background WT and IPKO mice were challenged intranasally with RSV A2001/2-20 strain. Bronchoalveolar lavage (BAL) fluid and lung were harvested after 12 and 24 hours. Control mice were not challenged with RSV. IFN-a and b protein expression in the BAL fluid and lung homogenate were measured by ELISA. RESULTS: IPKO mice had significantly greater BAL IFN-a and b protein expression compared to WT mice (p<0.05) 12 hours after intranasal challenge of RSV. Lung IFN-a protein expression was increased in IPKO mice compared to WT mice (p<0.05), but IFN-b was not significantly different. There was a trend for a difference of both BAL and lung IFN-a and b protein expression 24 hours after RSV challenge. CONCLUSIONS: IPKO mice had significantly increased IFN-a and b protein expression in the airway and lung. This result suggests that endogenous PGI2 signaling down-regulates RSV-induced illness through suppression of type I IFN induction.
682
Rhinovirus Induces Differential Expression of Th2-Promoting Epithelial Cytokines From Asthmatics Ex Vivo Joshua L. Kennedy, MD1, Larry Borish, MD, FAAAAI2, Peter W. Heymann, MD1, John W. Steinke, PhD, FAAAAI2; 1University of Virginia, Charlottesville, VA, 2Asthma and Allergic Disease Center, Carter Center for Immunology Research, University of Virginia, Charlottesville, VA. RATIONALE: The majority of childhood asthma exacerbations are provoked by rhinovirus (RV) infections, and these most often occur when allergies are also present. We propose this reflects the propensity of RV infection of epithelium to induce expression of Th2-promoting cytokines and that this occurs without altering the innate immune response of these cells to RV. METHODS: Epithelial cells derived from sinonasal epithelium of asthmatics and non-asthmatics were cultured and infected with RV39. Cells and supernatants were collected at various time intervals. Quantitative PCR was used to measure gene expression for the Th2-inducing cytokines IL-25, IL-33, and TSLP, and innate immune cytokines IL-15, IFN-ß, and IL-18. Supernatants were assayed for cytokine protein secretion. RESULTS: mRNA levels for IL-25, IL-33, and TSLP were increased in asthmatics compared to non-asthmatics after infection with RV39 at 2 (IL25), 4 (TSLP), and 24 hours (IL-33). mRNA levels were increased for IL15, IFN-ß, and IL-18 though no differences were observed between asthmatics and non-asthmatics. Protein expression of IL-15 was comparably evident in both cohorts at 48 hours. CONCLUSIONS: In contrast to non-asthmatics, RV-infected sinonasal epithelium derived from asthmatics express mRNA for cytokines important in the induction of Th2 immune deviation. Additionally, there was little difference in the expression of innate cytokines, specifically IL-15 and IL-18, which suggests that there is no defect in innate immune responses to RV amongst asthmatics. We propose instead that it is this RVinduced barrage of Th2-inducing cytokines that is responsible for asthma exacerbations.
683
Genetic Variants in Thymic Stromal Lymphopoietin (TSLP) and Receptor (TSLPR) and Their Influence On the Humoral Immune Response Luis Fang, Cesar Mu~noz, Luz Hernandez, Beatriz Martinez, Javier Marrugo; University of Cartagena, Cartagena, Colombia. RATIONALE: TSLP/TSLPR participates in the initiation of the Th2immune response. Recently, genetic variants of these genes have been associated with atopic diseases. However, it’s unknown whether these polymorphisms influence humoral immune response. This study evaluated the influence of polymorphisms in TSLP and TSLPR on the antibody response against dietary antigens, Blomia tropicalis and Ascaris lumbricoides. METHODS: By RT-qPCR and TaqManÒ probes, we genotyped the SNPs: rs1837253 [C/T], rs17551370 [A/G], rs2289276 [C/T] in TSLP and rs36139698 [A/G], rs36177645 [C/T], and rs36133495 [A/G] in TSLPR in 80 African-descent individuals. The serum levels of specific IgE, IgA and IgG4 against Blomia tropicalis, egg, milk, and peanut extracts and IgE against Ascaris lumbricoides extract, were measured by ELISA. Data analysis was performed using principal component analysis (PCA) by R Project software. RESULTS: The minor allele frequencies were as following: rs1837253 [T:0.41.9%], rs7551370 [A:14.4%], rs2289276 [T:22.5%], rs36139698 [A:37%], rs36177645 [C:24%] and rs36133495 [G:45%]. The PCA showed that specific IgE-IgG4 responses to dietary antigens were different from B. tropicalis response, while specific IgA response was similar in both groups of antigens, furthermore IgE response against B. tropicalis and A. lumbricoides were positively correlated. The genotype CT/TT in rs1837253 influenced IgE-IgG4 responses independent of antigenic group, while GG in rs17551370, CT/TT in rs2289276, AG/GG in rs36139698 and CT/TT in rs36177645 strongly favor IgE-IgG4 responses to egg and peanuts extracts. Only AG/GG in rs36133495 influenced IgA response against dietary and B. tropicalis antigens. CONCLUSIONS: TSLP/TSLPR variants influence the IgE-IgG4-IgA immune responses against dietary and environment antigens in African-descent individuals from Colombia.
684
Immune Modulatory Effects of IL-22 On Allergen-Induced Pulmonary Inflammation Ping Fang; Asthma & allergy center, Johns Hopkins University School of Medicine. RATIONALE: Allergen-induced pulmonary inflammation in asthma is mainly TH2 dominated. A recently discovered TH17/TH22 cell-derived cytokine, IL-22, has been found to be increased in asthma. However, several studies showed controversial findings in the immune modulatory effects of IL-22 in animal models of allergic asthma. Our objective was to determine the regulatory role of IL-22 in OVA-induced allergic inflammation using a novel lung-specific IL-22 transgenic overexpression system. METHODS: Inducible IL-22 transgenic (CC10-rtTA- and SPC-rtTA-TREtight-IL-22) mice on C57BL/6 genetic background were generated. IHC of lung tissue and ELISA of bronchoalveolar lavage (BAL) fluid for IL-22 were performed to confirm the location and quantity of IL-22 expression. OVAinduced pulmonary inflammatory responses were compared between IL-22 transgenic and wild type control mice. BAL total and differential cell counts, lung histology, mucous metaplasia, IHC of major basic protein for eosinophils, and serum OVA-specific IgE were determined. RESULTS: IL-22 transgene was successfully activated in the large (CC10 promoter) and small (SPC promoter) airway epithelium cells by doxycycline in the drinking water. OVA sensitization and challenge did not induce endogenous IL-22 expression. Compared to wild type mice, IL-22 transgenic mice showed decreased percentage of eosinophils in the BAL and slight reduction in eosinophilic inflammation and mucus metaplasia in the airways. Interestingly, inflammatory cell infiltration in lung parenchyma appeared slightly accentuated. In addition, there was no statistic difference in serum OVA-specific IgE. CONCLUSIONS: These findings indicate that IL-22 may have immune modulatory effects on pulmonary inflammatory responses in the development of allergen-induced asthma.
MONDAY
681