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Effect of macrophage inflammatory protein-1α on airway inflammation of asthma in mouse model
Inhibition of Human Cyclooxygenase-1 by Dermatophagoides Allergenic Extracts M. Sanchez-Borges1, M. Ouellet2, M. Percival3, A. Capriles-Hulett1, F. Caballero-Fonseca1; 1Allergy and Immunology, Centro Médico-Docente La Trinidad, Caracas, VENEZUELA, 2Merck Frosst Canada & Co., Kirkland, PQ, CANADA, 3Allergy and Immunology, Merck Frosst Canada & Co., Kirkland, PQ, CANADA. RATIONALE: An increased prevalence of cutaneous NSAID hypersensitivity reactions (urticaria and/or angioedema) has been observed in patients who experience systemic anaphylaxis triggered by mite-contaminated foods. We have investigated the presence of substances that inhibit human cyclooxygenases (COX) in Dermatophagoides allergenic extracts. METHODS: The in vitro effects of Dermatophagoides farinae and Dermatophagoides pteronyssinus commercial allergenic extracts on COX-1 and COX-2 isoenzymes were studied using recombinant human COX-1 and COX-2 microsomal assays. RESULTS: COX-1 enzymatic activity was inhibited by a 2000 AU/ml concentration of D. farinae and D. pteronyssinus extracts by 76 and 70 %, respectively. On the other hand, both extracts produced a significant activation of COX-2 activity at concentrations above 220 AU/ml. CONCLUSIONS: COX-1 inhibition by mite-derived products may contribute to the clinical picture of severe adverse reactions observed in some NSAID-sensitive subjects occurring immediately after oral/GI mucosal exposure to foods contaminated with mites.
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RV1A and dsRNA Highly Induce a Biphasic Expression of MIP-3/CCL20 in Human Bronchial Epithelial Cells (hBECs) J. M. Bellak1, K. Hanson2, R. A. Brockman-Schneider2, H. Dagher2, J. E. Gern2; 1Medicine, University of Wisconsin Hospital & Clinics, Madison, WI, 2Pediatrics, University of Wisconsin Hospital & Clinics, Madison, WI. RATIONALE: MIP-3 (CC family chemokine also known as CCL20) is a potent chemoattractant for naïve dendritic cells and memory lymphocytes. Rhinovirus infections in human respiratory epithelium were found to induce CCL20 mRNA by gene expression profiling, and CCL20 secretion during the acute phase of an experimental cold. Mechanisms for upregulation of CCL20 by viruses are not well understood. METHODS: To define the mechanisms of CCL20 induction in hBECs, cell monolayers were incubated with RV1A and dsRNA; CCL20 mRNA was measured by qPCR and protein secretion by ELISA. RESULTS: Analysis of CCL20 mRNA induction by expression profiling and qPCR yielded similar results. hBECs stimulated with RV1A induced a biphasic pattern of CCL20 mRNA (4.9-, 1.9-, and 4.9-fold at 4, 8, and 16 hours; p<0.05). dsRNA induced greater amounts of CCL20 mRNA, with a similar pattern (34-, 9.8-, and 49-fold at 4, 8, and 16 hours; p<0.05). Unstimulated hBECs secreted small amounts of CCL20 (224 pg/ml after 48 hours incubation), and this was increased 1.9-fold by RV1A. dsRNA induced CCL20 secretion more rapidly (2.9-fold increase after 16 hours incubation). CONCLUSIONS: RV1A and dsRNA are potent inducers of CCL20 expression in hBECs. The biphasic pattern of induction suggests both receptor binding and replication processes may contribute to CCL20 upregulation. CCL20 effects on mononuclear cell recruitment suggest a potential role in antiviral responses. Funding: NIH grants M01 RR03186, P01 HL70831, and P01 AI50500
A Comparison of Cytokine Responses in Respiratory Syncytial Virus and Adenovirus Infections In Vitro J. S. Yoon, M. H. Lee, H. H. Kim, J. T. Kim, J. S. Lee; Department of pediatrics, the Catholic University of Korea, Seoul, REPUBLIC OF KOREA. RATIONALE: Respiratory syncytial virus(RSV) and adenovirus(AV) infections are indistinguishable during the acute phases of the diseases. However, long-term prognosis is different. RSV infection is related to later development of asthma, and AV, mainly AV serotype 3, 7 and 21, is related to residual airway damage manifested by bronchiectasis, bronchiolitis obliterans. We hypothesized that this difference could be partly due to different immune responses induced by these viruses. METHODS: To test this hypothesis we infected human bronchial epithelial cell line BEAS-2B with RSV and AV serotype 3 at the same multiplicity of infection 1, and then the total RNAs and the supernatants were obtained at 24 hour. BEAS-2B cells uninfected were used as a control. IL6, IL-8 and RANTES mRNA expression and protein production were determined by RT-PCR and ELISA respectively. RESULTS: BEAS-2B cells showed mRNA expression and production of IL-6, IL-8, and RANTES constitutively and after RSV or AV infection. The cells infected with RSV showed a significant increase in the production of IL-6, IL-8, and RANTES protein(P<0.05) and with AV showed a significant increase in the production of IL-6 and RANTES protein(P<0.05) but not of IL-8. IL-6, IL-8, and RANTES protein in cells with RSV infection were significantly higher than those with AV infection(P<0.05). CONCLUSIONS: These results suggest that there may be different virus-specific induction of IL-6, IL-8, and RANTES production during infection of airway epithelial cells. Because IL-6, IL-8, and RANTES are the important mediators of airway inflammation, virus-specific induction of these cytokines may explain the different long-term prognosis of airway infection with different virus. Funding: the Catholic University of Korea
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Effect of Macrophage Inflammatory Protein-1 on Airway Inflammation of Asthma in Mouse Model C. Li, W. Zhang, X. Chen, L. Xie, Q. He, X. Hu, J. Li, M. Li, R. Wu, Z. Zhang; Division of Pulmonology, Yuying Children’s Hospital, Wenzhou Medical College, Wenzhou City, Zhejiang, P.R. China., CHINA. RATIONALE: Macrophage inflammatory protein-1 (MIP-1) is involved in airway inflammation of mouse asthma. METHODS: Mouse asthma was established by the ovalbumin (OVA) sensitization/challenge method. Seventy male BALB/C mice were randomly divided into control and asthma groups that received vehicle and OVA respectively. At different time points after last challenge, sera and bronchoalveolar lavage fluid (BALF) were collected for MIP-1 protein level determination using ELISA. The total cell counts and cell distribution was also determined in BALF. MIP-1 protein positive cells were identified by immunohistochemistry, mRNA by in situ hybridization. RESULTS: At 24 h after last challenge, MIP-1 level in BALF and sera of asthma group (42.9 ± 5.8 pg/ml and 41.7 ± 6.3 pg/ml, n = 10) were significant higher than control group (20.9 ± 3.8 pg/ml and 22.4 ± 4.3pg/ml, P<0.01). This increase was noticed as early as 3 h after challenge and peaked at 24 h. Immunohistochemistry showed that MIP-1 positive cells at the bronchus were elevated in asthma group (26.4 ± 6.2 %) as compared to control group (10.3 ± 2.5 %, P<0.01). It was mainly expressed in epithelial cells. In situ hybridization showed that MIP-1 mRNA expression around the bronchus of asthma animals (23.9 ± 4.2 %) were significantly increased when compared to those of control (8.7 ± 1.8 %, P<0.01). Again, they were mainly located within epithelial cells. Result further demonstrates strong correlation between MIP-1 level and the total and percentage of EOS in BALF. CONCLUSIONS: Epithelial produced MIP-1 plays an important role in animal model of asthma. Funding: Zhejiang provincial educational foundation,P.R. China
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Abstracts S51
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2
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RV1A and dsRNA Highly Induce a Biphasic Expression of MIP-3/CCL20 in Human Bronchial Epithelial Cells (hBECs) J. M. Bellak1, K. Hanson2, R. A. Brockman-Schneider2, H. Dagher2, J. E. Gern2; 1Medicine, University of Wisconsin Hospital & Clinics, Madison, WI, 2Pediatrics, University of Wisconsin Hospital & Clinics, Madison, WI. RATIONALE: MIP-3 (CC family chemokine also known as CCL20) is a potent chemoattractant for naïve dendritic cells and memory lymphocytes. Rhinovirus infections in human respiratory epithelium were found to induce CCL20 mRNA by gene expression profiling, and CCL20 secretion during the acute phase of an experimental cold. Mechanisms for upregulation of CCL20 by viruses are not well understood. METHODS: To define the mechanisms of CCL20 induction in hBECs, cell monolayers were incubated with RV1A and dsRNA; CCL20 mRNA was measured by qPCR and protein secretion by ELISA. RESULTS: Analysis of CCL20 mRNA induction by expression profiling and qPCR yielded similar results. hBECs stimulated with RV1A induced a biphasic pattern of CCL20 mRNA (4.9-, 1.9-, and 4.9-fold at 4, 8, and 16 hours; p<0.05). dsRNA induced greater amounts of CCL20 mRNA, with a similar pattern (34-, 9.8-, and 49-fold at 4, 8, and 16 hours; p<0.05). Unstimulated hBECs secreted small amounts of CCL20 (224 pg/ml after 48 hours incubation), and this was increased 1.9-fold by RV1A. dsRNA induced CCL20 secretion more rapidly (2.9-fold increase after 16 hours incubation). CONCLUSIONS: RV1A and dsRNA are potent inducers of CCL20 expression in hBECs. The biphasic pattern of induction suggests both receptor binding and replication processes may contribute to CCL20 upregulation. CCL20 effects on mononuclear cell recruitment suggest a potential role in antiviral responses. Funding: NIH grants M01 RR03186, P01 HL70831, and P01 AI50500
A Comparison of Cytokine Responses in Respiratory Syncytial Virus and Adenovirus Infections In Vitro J. S. Yoon, M. H. Lee, H. H. Kim, J. T. Kim, J. S. Lee; Department of pediatrics, the Catholic University of Korea, Seoul, REPUBLIC OF KOREA. RATIONALE: Respiratory syncytial virus(RSV) and adenovirus(AV) infections are indistinguishable during the acute phases of the diseases. However, long-term prognosis is different. RSV infection is related to later development of asthma, and AV, mainly AV serotype 3, 7 and 21, is related to residual airway damage manifested by bronchiectasis, bronchiolitis obliterans. We hypothesized that this difference could be partly due to different immune responses induced by these viruses. METHODS: To test this hypothesis we infected human bronchial epithelial cell line BEAS-2B with RSV and AV serotype 3 at the same multiplicity of infection 1, and then the total RNAs and the supernatants were obtained at 24 hour. BEAS-2B cells uninfected were used as a control. IL6, IL-8 and RANTES mRNA expression and protein production were determined by RT-PCR and ELISA respectively. RESULTS: BEAS-2B cells showed mRNA expression and production of IL-6, IL-8, and RANTES constitutively and after RSV or AV infection. The cells infected with RSV showed a significant increase in the production of IL-6, IL-8, and RANTES protein(P<0.05) and with AV showed a significant increase in the production of IL-6 and RANTES protein(P<0.05) but not of IL-8. IL-6, IL-8, and RANTES protein in cells with RSV infection were significantly higher than those with AV infection(P<0.05). CONCLUSIONS: These results suggest that there may be different virus-specific induction of IL-6, IL-8, and RANTES production during infection of airway epithelial cells. Because IL-6, IL-8, and RANTES are the important mediators of airway inflammation, virus-specific induction of these cytokines may explain the different long-term prognosis of airway infection with different virus. Funding: the Catholic University of Korea
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Effect of Macrophage Inflammatory Protein-1 on Airway Inflammation of Asthma in Mouse Model C. Li, W. Zhang, X. Chen, L. Xie, Q. He, X. Hu, J. Li, M. Li, R. Wu, Z. Zhang; Division of Pulmonology, Yuying Children’s Hospital, Wenzhou Medical College, Wenzhou City, Zhejiang, P.R. China., CHINA. RATIONALE: Macrophage inflammatory protein-1 (MIP-1) is involved in airway inflammation of mouse asthma. METHODS: Mouse asthma was established by the ovalbumin (OVA) sensitization/challenge method. Seventy male BALB/C mice were randomly divided into control and asthma groups that received vehicle and OVA respectively. At different time points after last challenge, sera and bronchoalveolar lavage fluid (BALF) were collected for MIP-1 protein level determination using ELISA. The total cell counts and cell distribution was also determined in BALF. MIP-1 protein positive cells were identified by immunohistochemistry, mRNA by in situ hybridization. RESULTS: At 24 h after last challenge, MIP-1 level in BALF and sera of asthma group (42.9 ± 5.8 pg/ml and 41.7 ± 6.3 pg/ml, n = 10) were significant higher than control group (20.9 ± 3.8 pg/ml and 22.4 ± 4.3pg/ml, P<0.01). This increase was noticed as early as 3 h after challenge and peaked at 24 h. Immunohistochemistry showed that MIP-1 positive cells at the bronchus were elevated in asthma group (26.4 ± 6.2 %) as compared to control group (10.3 ± 2.5 %, P<0.01). It was mainly expressed in epithelial cells. In situ hybridization showed that MIP-1 mRNA expression around the bronchus of asthma animals (23.9 ± 4.2 %) were significantly increased when compared to those of control (8.7 ± 1.8 %, P<0.01). Again, they were mainly located within epithelial cells. Result further demonstrates strong correlation between MIP-1 level and the total and percentage of EOS in BALF. CONCLUSIONS: Epithelial produced MIP-1 plays an important role in animal model of asthma. Funding: Zhejiang provincial educational foundation,P.R. China
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Abstracts S51
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2