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Gamma Tocopherol Inhibits Ozone-Induced Inflammatory Gene Expression in Airway Epithelial Cells
AB262 Abstracts
821
Cat Dander Extract Directly Stimulates B Cell to Induce Class Switch
Toshiko Itazawa, MD, PhD, Koa Hosoki, MD, PhD, and Sanjiv Sur, MD; University of Texas Medical Branch, Galveston, TX. RATIONALE: Prior reports have indicated that low molecular weight pollen-derived factors such as phytoprostane E1 can directly stimulate class switch of B-cells to IgE secreting cells. Cat dander is a potent allergen that sensitizes a large percentage of the population. However the ability of cat dander extract (CDE) to directly stimulate IgE class switch has not been reported. We hypothesized that CDE can also directly stimulate na€ıve Bcell class switch to IgE in the presence of T-cell help and IL-4. METHODS: Purified B cells from the spleen of wild type (WT) C57BL/6 mice or Myd88KO mice were cultured with CDE in the presence of IL-4 and/or anti-CD40 antibody. Total IgE in cell culture supernatants were measured by ELISA. Real-time PCR were performed to assess IgE class switch. RESULTS: In the presence of IL-4 and anti-CD40 antibody, CDE provided a strong signal to vigorously promote IgE class switch in na€ıve B cells from WT mice, and increased expression of Nfil3, GLε (germ-line ε) and GLg (germ-line g). Disruption of Myd88 in B-cells blocked CDEstimulated IgE production and the increase in mRNA expression of Nfil3, GLε and GLg. CONCLUSIONS: CDE directly stimulates B cells to induce IgE class switch through a Myd88-dependent signaling mechanism in the presence of a IgE switch factor (IL-4) and T-cell help (anti-CD40).
822
Relation of hepatitis B virus infection in Brooklyn immigrants to allergic responses and asthma
MONDAY
Tehila A. Saadia, MD1, Maria-Anna Vastardi, MD1,2, Maria A. Chitty Lopez, MD3, Irina Katayeva, MD1, Stephan Kohlhoff, MD3, and Rauno Joks, MD, FAAAAI1; 1Center for Allergy and Asthma Research at SUNY Downstate Medical Center, Brooklyn, NY, 2Lutheran Medical Center, Brooklyn, NY, 3SUNY Downstate Dept of Pediatrics, Brooklyn, NY. RATIONALE: Hepatitis B virus (HBV) is an immunomodulatory virus and has been linked to IgE production. We determined the association of HBV serologies to IgE responses in Brooklyn immigrants from HBVendemic countries. METHODS: Serology testing (ELISA, Abnova) was performed on immigrants with (n5167) and without (n5175) asthma, allergic rhinitis, and food allergies. Patients who tested positive for HBV surface antibody (anti-HBs) were tested for HBV core antibody (anti-HBc) to identify natural infection versus vaccination. Anti-HBs negative subjects were further tested for HBV surface antigen (HBsAg) to identify infection prior to seroconversion. Serum IgE levels and exhaled nitric oxide (eNO) (Niox, Aerocrine) measurements were obtained on all patients. Chi-square tests were performed for associations between HBV groups [(1)non-vaccinated, non-infected; (2)vaccinated only; (3)past/current infection] and allergic diseases. Kruskal-Wallis tests were performed to compare distribution of serum IgE and eNO between HBV groups. RESULTS: 66% (n5226/342) of the sampled Brooklyn immigrant patient population was found to have past or current HBV infection. The prevalence of allergic diseases in HBV infected patients was 50.4% (n5114). We did not find significant association between natural HBV infection and development of seasonal allergies (p50.39), asthma (p50.25), food allergies (p50.15), IgE (p50.59), or eNO (p50.24). Furthermore, there was no association of these factors with vaccination or non-infected/non-vaccinated status (p5NS). CONCLUSIONS: There is no significant association between HBV infection and the development of IgE responses in this Brooklyn immigrant patient cohort.
J ALLERGY CLIN IMMUNOL FEBRUARY 2017
823
Gamma Tocopherol Inflammatory Gene Epithelial Cells
Inhibits Ozone-Induced Expression in Airway
Charity G. Duran, PhD1, and Michelle L. Hernandez, MD2; 1University of North Carolina, Chapel Hill, NC, 2University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC. RATIONALE: Allergic airway inflammation can be further exacerbated by exposure to environmental pollutants such as ozone (O3). g-Tocopherol (gT), the primary dietary form of Vitamin E, possesses an array of anti-inflammatory properties; because airway epithelial cells are direct targets of O3-induced injury, we sought to investigate the potential anti-inflammatory effects of gT and one of its major metabolites, g-CEHC, on airway epithelial cells using an in vitro O3-exposure model. METHODS: Immortalized human bronchial epithelial cells (16HBEs), were grown at air liquid interface and exposed to 0.4ppm O3 for 4 hours with a 1 hour recovery. Cells were incubated with DMSO or gT overnight, or g-CEHC 1 hour prior to exposure. Fold changes in inflammatory gene expression were quantified using qRT-PCR. Statistically significant differences were determined by one way ANOVA with a Kruskal-Wallis posttest. RESULTS: O3-exposure, in comparison with clean air-exposure, significantly increased expression of IL-6 (4.3 6 0.8 vs 0.9 6 0.1, p50.005) and IL-8 (4.1 6 0.7 vs 0.9 6 0.1, p50.005) in DMSO vehicle controltreated cells. Treatment with either 40mM g-CEHC or gT blunted O3induced expression of IL-8 compared to clean air (2.5 6 0.1 vs 1.0 6 0.2, p50.2 and 2.1 6 0.5 vs 1.3 6 0.07, p50.85 respectively). CONCLUSIONS: This study is the first to demonstrate that gT can abrogate inflammatory gene expression in airway epithelial cells in response to O3-exposure. Epithelial-derived IL-8 is integral for recruitment of neutrophils in response to airway injury, suggesting that gT is a candidate therapeutic for reducing neutrophilic airway inflammation.
824
Gastric Juice Directly Enhanced IL-13-Induced CCL26 Expression in Human Bronchial Epithelial Cells in Vitro
Keisuke Orimo, MD1,2, Kyoko Futamura, MD, PhD1, Naoko Okada, PhD1, and Kenji Matsumoto, MD, PhD1; 1Department of Allergy and Clinical Immunology, National Research Institute for Child Health and Development, Tokyo, Japan, 2First Department of Medicine, Tokyo Women’s Medical University School of Medicine, Tokyo, Japan. RATIONALE: Gastroesophageal reflux disease (GERD) reportedly correlates with difficult asthma, but the mechanisms of how GERD affects asthma pathogenesis remain obscure. In the present study, we investigated the direct effects of gastric juice on chemokine expression in human bronchial epithelial cells in vitro. METHODS: An immortalized human bronchial epithelial cell line (BEAS-2B) and normal human bronchial epithelial cells (NHBE) were exposed to acidic medium (pH 2) alone or with pepsin (gastric juice), followed by culture in normal medium in the presence or absence of IL-13. CCL26 (eotaxin-3) mRNA in the cell lysates and its protein in the culture supernatants were measured by qPCR and ELISA, respectively. RESULTS: In both BEAS-2B and NHBE, gastric juice induced significant CCL26 mRNA but its protein was undetectable, whereas the acidic medium alone showed much weaker effects. The gastric juice showed significant synergistic enhancement of IL-13-induced CCL26 mRNA and protein production. Gastric juice did not induce phosphorylation of intracellular STAT6 in BEAS-2B. CONCLUSIONS: Our results suggest that microaspiration of gastric juice directly exacerbates type 2 inflammation through enhancement of CCL26 production in the airway epithelial cells, perhaps leading to asthma exacerbation.
821
Cat Dander Extract Directly Stimulates B Cell to Induce Class Switch
Toshiko Itazawa, MD, PhD, Koa Hosoki, MD, PhD, and Sanjiv Sur, MD; University of Texas Medical Branch, Galveston, TX. RATIONALE: Prior reports have indicated that low molecular weight pollen-derived factors such as phytoprostane E1 can directly stimulate class switch of B-cells to IgE secreting cells. Cat dander is a potent allergen that sensitizes a large percentage of the population. However the ability of cat dander extract (CDE) to directly stimulate IgE class switch has not been reported. We hypothesized that CDE can also directly stimulate na€ıve Bcell class switch to IgE in the presence of T-cell help and IL-4. METHODS: Purified B cells from the spleen of wild type (WT) C57BL/6 mice or Myd88KO mice were cultured with CDE in the presence of IL-4 and/or anti-CD40 antibody. Total IgE in cell culture supernatants were measured by ELISA. Real-time PCR were performed to assess IgE class switch. RESULTS: In the presence of IL-4 and anti-CD40 antibody, CDE provided a strong signal to vigorously promote IgE class switch in na€ıve B cells from WT mice, and increased expression of Nfil3, GLε (germ-line ε) and GLg (germ-line g). Disruption of Myd88 in B-cells blocked CDEstimulated IgE production and the increase in mRNA expression of Nfil3, GLε and GLg. CONCLUSIONS: CDE directly stimulates B cells to induce IgE class switch through a Myd88-dependent signaling mechanism in the presence of a IgE switch factor (IL-4) and T-cell help (anti-CD40).
822
Relation of hepatitis B virus infection in Brooklyn immigrants to allergic responses and asthma
MONDAY
Tehila A. Saadia, MD1, Maria-Anna Vastardi, MD1,2, Maria A. Chitty Lopez, MD3, Irina Katayeva, MD1, Stephan Kohlhoff, MD3, and Rauno Joks, MD, FAAAAI1; 1Center for Allergy and Asthma Research at SUNY Downstate Medical Center, Brooklyn, NY, 2Lutheran Medical Center, Brooklyn, NY, 3SUNY Downstate Dept of Pediatrics, Brooklyn, NY. RATIONALE: Hepatitis B virus (HBV) is an immunomodulatory virus and has been linked to IgE production. We determined the association of HBV serologies to IgE responses in Brooklyn immigrants from HBVendemic countries. METHODS: Serology testing (ELISA, Abnova) was performed on immigrants with (n5167) and without (n5175) asthma, allergic rhinitis, and food allergies. Patients who tested positive for HBV surface antibody (anti-HBs) were tested for HBV core antibody (anti-HBc) to identify natural infection versus vaccination. Anti-HBs negative subjects were further tested for HBV surface antigen (HBsAg) to identify infection prior to seroconversion. Serum IgE levels and exhaled nitric oxide (eNO) (Niox, Aerocrine) measurements were obtained on all patients. Chi-square tests were performed for associations between HBV groups [(1)non-vaccinated, non-infected; (2)vaccinated only; (3)past/current infection] and allergic diseases. Kruskal-Wallis tests were performed to compare distribution of serum IgE and eNO between HBV groups. RESULTS: 66% (n5226/342) of the sampled Brooklyn immigrant patient population was found to have past or current HBV infection. The prevalence of allergic diseases in HBV infected patients was 50.4% (n5114). We did not find significant association between natural HBV infection and development of seasonal allergies (p50.39), asthma (p50.25), food allergies (p50.15), IgE (p50.59), or eNO (p50.24). Furthermore, there was no association of these factors with vaccination or non-infected/non-vaccinated status (p5NS). CONCLUSIONS: There is no significant association between HBV infection and the development of IgE responses in this Brooklyn immigrant patient cohort.
J ALLERGY CLIN IMMUNOL FEBRUARY 2017
823
Gamma Tocopherol Inflammatory Gene Epithelial Cells
Inhibits Ozone-Induced Expression in Airway
Charity G. Duran, PhD1, and Michelle L. Hernandez, MD2; 1University of North Carolina, Chapel Hill, NC, 2University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC. RATIONALE: Allergic airway inflammation can be further exacerbated by exposure to environmental pollutants such as ozone (O3). g-Tocopherol (gT), the primary dietary form of Vitamin E, possesses an array of anti-inflammatory properties; because airway epithelial cells are direct targets of O3-induced injury, we sought to investigate the potential anti-inflammatory effects of gT and one of its major metabolites, g-CEHC, on airway epithelial cells using an in vitro O3-exposure model. METHODS: Immortalized human bronchial epithelial cells (16HBEs), were grown at air liquid interface and exposed to 0.4ppm O3 for 4 hours with a 1 hour recovery. Cells were incubated with DMSO or gT overnight, or g-CEHC 1 hour prior to exposure. Fold changes in inflammatory gene expression were quantified using qRT-PCR. Statistically significant differences were determined by one way ANOVA with a Kruskal-Wallis posttest. RESULTS: O3-exposure, in comparison with clean air-exposure, significantly increased expression of IL-6 (4.3 6 0.8 vs 0.9 6 0.1, p50.005) and IL-8 (4.1 6 0.7 vs 0.9 6 0.1, p50.005) in DMSO vehicle controltreated cells. Treatment with either 40mM g-CEHC or gT blunted O3induced expression of IL-8 compared to clean air (2.5 6 0.1 vs 1.0 6 0.2, p50.2 and 2.1 6 0.5 vs 1.3 6 0.07, p50.85 respectively). CONCLUSIONS: This study is the first to demonstrate that gT can abrogate inflammatory gene expression in airway epithelial cells in response to O3-exposure. Epithelial-derived IL-8 is integral for recruitment of neutrophils in response to airway injury, suggesting that gT is a candidate therapeutic for reducing neutrophilic airway inflammation.
824
Gastric Juice Directly Enhanced IL-13-Induced CCL26 Expression in Human Bronchial Epithelial Cells in Vitro
Keisuke Orimo, MD1,2, Kyoko Futamura, MD, PhD1, Naoko Okada, PhD1, and Kenji Matsumoto, MD, PhD1; 1Department of Allergy and Clinical Immunology, National Research Institute for Child Health and Development, Tokyo, Japan, 2First Department of Medicine, Tokyo Women’s Medical University School of Medicine, Tokyo, Japan. RATIONALE: Gastroesophageal reflux disease (GERD) reportedly correlates with difficult asthma, but the mechanisms of how GERD affects asthma pathogenesis remain obscure. In the present study, we investigated the direct effects of gastric juice on chemokine expression in human bronchial epithelial cells in vitro. METHODS: An immortalized human bronchial epithelial cell line (BEAS-2B) and normal human bronchial epithelial cells (NHBE) were exposed to acidic medium (pH 2) alone or with pepsin (gastric juice), followed by culture in normal medium in the presence or absence of IL-13. CCL26 (eotaxin-3) mRNA in the cell lysates and its protein in the culture supernatants were measured by qPCR and ELISA, respectively. RESULTS: In both BEAS-2B and NHBE, gastric juice induced significant CCL26 mRNA but its protein was undetectable, whereas the acidic medium alone showed much weaker effects. The gastric juice showed significant synergistic enhancement of IL-13-induced CCL26 mRNA and protein production. Gastric juice did not induce phosphorylation of intracellular STAT6 in BEAS-2B. CONCLUSIONS: Our results suggest that microaspiration of gastric juice directly exacerbates type 2 inflammation through enhancement of CCL26 production in the airway epithelial cells, perhaps leading to asthma exacerbation.