- Email: [email protected]
B Cell Activating Factor of the TNF Family (BAFF) in Allergic Bronchopulmonary Aspergillosis (ABPA) and Cystic Fibrosis (CF)
S204 Abstracts
783
B Cell Activating Factor of the TNF Family (BAFF) in Allergic Bronchopulmonary Aspergillosis (ABPA) and Cystic Fibrosis (CF) P. A. Greenberger, A. Kato; Northwestern University Feinberg School of Medicine, Chicago, IL. RATIONALE: Since total and anti-A. fumigatus IgE and IgG can be increased or normal depending on disease activity, we measured serum BAFF in ABPA and CF. We hypothesized that BAFF would be elevated in sera from patients with ABPA compared to patients with fungal (A.fumigatus 1) asthma. METHODS: Serum BAFF was measured by ELISA (R & D Systems, Minneapolis, MN) with lowest concentration of detection of 31 pg/ml. Sera were from pooled sera from non-atopic patients, patients with A.fumigutus 1 asthma without ABPA, individual ABPA, and cystic fibrosis sera (with sensitization to A.fumigatus but low total serum IgE concentrations). RESULTS: Serum BAFF was elevated in pooled sera from ABPA (1150 pg/ml) compared to pooled sera from A.fumigatus 1 asthma (817 pg/ml) and sera from non-atopics (883 pg/ml). ABPA sera (n 513; 945 6 407 pg/ml) were less than non-ABPA CF sera (n 5 8:1851 6 849 pg/ml), p < 0.01. To explore the relationship between BAFF and total IgE in ABPA (n 5 13, total IgE 2970 6 1280 kU/L) vs. CF without ABPA (n 5 8, total IgE 181 6 224 kU/L), we selected sera with equally strongly positive serum anti-A.fumigatus IgE and IgG antibodies. There was an inverse relationship between total serum IgE and serum BAFF (Spearman -0.574, p < 0.01) despite elevated anti A.fumigatus antibodies. CONCLUSIONS: 1. Serum BAFF was elevated in ABPA as compared to A.fumigatus 1 asthma and controls. 2. BAFF was greater in sera from CF compared to ABPA despite much lower total IgE concentrations in CF. 3. BAFF was inversely correlated with the total IgE concentration. Funding: Ernest S. Bazley Grant to Northwestern University and Northwestern Memorial Hospital and RO1 AI072570
MONDAY
784
In Vitro Adverse Effects of Corticosteroid on TNF-a-mediated Responses by Pulmonary Microvascular Endothelial Cells K. Orihara, S. Fukuda, H. Fujinaga, K. Matsumoto, H. Saito, A. Matsuda; National Research Institute for Child Health and Development, Tokyo, JAPAN. RATIONALE: Chronic severe asthma accompanied by a marked increase in TNF-a production might contribute to corticosteroid refractoriness, but its process is still unknown. TNF-a strongly activates endothelium, leading to the enhancement of local inflammation through leukocyte extravasation. We examined the effects of dexamethasone (DEX) on the TNF-a-treated human lung blood microvascular endothelial cells (HMVEC-LBl) compared with those of bronchial smooth muscle cells (BSMC) in vitro. METHODS: HMVEC-LBl and BSMC were3 cultured with TNF-a in the presence or absence of DEX. Cell surface expression of VCAM-1 and ICAM-1 was analyzed by flow cytometry. Chemokine expressions were determined by real-time PCR and ELISA. Apoptotic cells were determined by Annexin V and PI staining. RESULTS: TNF-a strongly induced VCAM-1 and ICAM-1 expression on HMVEC-LBl. Although DEX showed no effect on the TNF-a-induced ICAM-1, TNF-a-induced VCAM-1 was enhanced by DEX treatment. TNF-a markedly induced CXCL1, CXCL8, CXCL10 and CCL5 in both HMVEC-LBl and BSMC. All of the TNF-a-induced chemokines in BSMC were significantly attenuated by DEX, whereas TNF-a-induced CXCL1 and CXCL8 in HMVEC-LBl were not affected at all. TNF-ainduced CXCL10 and CCL5 in HMVEC-LBl were enhanced by DEX. Furthermore, TNF-a-induced apoptosis of HMVEC-LBl was blocked by DEX. CONCLUSIONS: Some unfavorable effects of corticosteroid were found in TNF-a-mediated in vitro reactions by HMVEC-LBl but not by BSMC, suggesting that extravasation of leukocytes and survival of endothelial cells may be upregulated in vivo. Corticosteroids should be given with
J ALLERGY CLIN IMMUNOL FEBRUARY 2008
caution to severe asthmatic patients with a marked increase in TNF-a production. Funding: National Institute of Biomedical Innovation
785
Quercetin Potentially Ameliorates the C5a-induced Acute Peritonitis J. J. Aarthi1, S. D. Kumar2, P. N. Pushparaj1; 1Department of Physiology, National University of Singapore, Singapore, SINGAPORE, 2Department of Anatomy, National University of Singapore, Singapore, SINGAPORE. RATIONALE: To investigate the effects of Quercetin in the anaphylatoxin C5a-triggered responses in vivo. Complement cascade activation leads to the generation of the potent proinflammatory anaphylatoxin, C5a. Significant amounts of C5a, as well as other complement products in the blood, can lead to a series of adverse effects associated with a variety of pathologies. METHODS: Acute inflammation in the peritoneal cavity was induced by an intraperitoneal (i.p.) injection of recombinant human C5a (2 mg per mouse in a final volume of 100 ml). Quercetin (75 mM/100 ml of PBS) was i.v. injected 10 min before C5a i.p. injection. Four mice were used for each group per experiment, and the experiments were conducted three times. The i.v. administration of C5a triggers a rapid reduction in neutrophil level but pre-treating mice with the Quercetin 10 min before the C5a i.v. administration substantially inhibited the neutropenia and increase in the serum levels of IL-6 and TNF-a. The C5a i.p. injection triggered a fast recruitment of neutrophils followed by monocytes into the peritoneal cavity. Extravasation due to the vascular permeability was also observed: RESULTS: In mice pretreated with Quercetin, there was a significant reduction on the C5a-triggered neutrophil and monocyte infiltration, and a marked reduction on the Evans blue influx. Immunohistochemical analysis had shown that the expression of cell adhesion molecules (CAMs) such as P-Selecin, ICAM-1, and VCAM-1 were significantly down-regulated in the peritoneal membrane of the Quercetin-treated mice. CONCLUSIONS: These observations suggest that Quercetin treatment potentially blocks the C5a-triggered inflammatory responses in vivo. Funding: National University of Singapore
786
Micro RNA-Directed Translation Repression Is DownRegulated in Activated Bronchial Epithelial Cells Y. Zhai; UT Medical School, Houston, TX. RATIONALE: Since activation of bronchial epithelial cells causes elevation of the levels of inflammation mediators and tissue-remodeling factors, we examined the role of micro RNA (miRNA)-directed translation repression in this regulation. METHODS: MicroRNAs in bronchial epithelial cells (BEAS-2B) were identified by microarray profiling. The effect of a representative miRNA on the protein synthesis of its target reporter mRNA was tested in BEAS-2B cells with or without TNF-a/IL-4 stimulation. RESULTS: Protein synthesis rate of the reporter mRNA carrying the miRNA target sites is higher in the cells treated with TNF-a/IL-4 than that in the non-treated cells. CONCLUSION: Allergic immune response in bronchial epithelial cells elicits down-regulation of miRNA-directed translation repression. Funding: Sandler
783
B Cell Activating Factor of the TNF Family (BAFF) in Allergic Bronchopulmonary Aspergillosis (ABPA) and Cystic Fibrosis (CF) P. A. Greenberger, A. Kato; Northwestern University Feinberg School of Medicine, Chicago, IL. RATIONALE: Since total and anti-A. fumigatus IgE and IgG can be increased or normal depending on disease activity, we measured serum BAFF in ABPA and CF. We hypothesized that BAFF would be elevated in sera from patients with ABPA compared to patients with fungal (A.fumigatus 1) asthma. METHODS: Serum BAFF was measured by ELISA (R & D Systems, Minneapolis, MN) with lowest concentration of detection of 31 pg/ml. Sera were from pooled sera from non-atopic patients, patients with A.fumigutus 1 asthma without ABPA, individual ABPA, and cystic fibrosis sera (with sensitization to A.fumigatus but low total serum IgE concentrations). RESULTS: Serum BAFF was elevated in pooled sera from ABPA (1150 pg/ml) compared to pooled sera from A.fumigatus 1 asthma (817 pg/ml) and sera from non-atopics (883 pg/ml). ABPA sera (n 513; 945 6 407 pg/ml) were less than non-ABPA CF sera (n 5 8:1851 6 849 pg/ml), p < 0.01. To explore the relationship between BAFF and total IgE in ABPA (n 5 13, total IgE 2970 6 1280 kU/L) vs. CF without ABPA (n 5 8, total IgE 181 6 224 kU/L), we selected sera with equally strongly positive serum anti-A.fumigatus IgE and IgG antibodies. There was an inverse relationship between total serum IgE and serum BAFF (Spearman -0.574, p < 0.01) despite elevated anti A.fumigatus antibodies. CONCLUSIONS: 1. Serum BAFF was elevated in ABPA as compared to A.fumigatus 1 asthma and controls. 2. BAFF was greater in sera from CF compared to ABPA despite much lower total IgE concentrations in CF. 3. BAFF was inversely correlated with the total IgE concentration. Funding: Ernest S. Bazley Grant to Northwestern University and Northwestern Memorial Hospital and RO1 AI072570
MONDAY
784
In Vitro Adverse Effects of Corticosteroid on TNF-a-mediated Responses by Pulmonary Microvascular Endothelial Cells K. Orihara, S. Fukuda, H. Fujinaga, K. Matsumoto, H. Saito, A. Matsuda; National Research Institute for Child Health and Development, Tokyo, JAPAN. RATIONALE: Chronic severe asthma accompanied by a marked increase in TNF-a production might contribute to corticosteroid refractoriness, but its process is still unknown. TNF-a strongly activates endothelium, leading to the enhancement of local inflammation through leukocyte extravasation. We examined the effects of dexamethasone (DEX) on the TNF-a-treated human lung blood microvascular endothelial cells (HMVEC-LBl) compared with those of bronchial smooth muscle cells (BSMC) in vitro. METHODS: HMVEC-LBl and BSMC were3 cultured with TNF-a in the presence or absence of DEX. Cell surface expression of VCAM-1 and ICAM-1 was analyzed by flow cytometry. Chemokine expressions were determined by real-time PCR and ELISA. Apoptotic cells were determined by Annexin V and PI staining. RESULTS: TNF-a strongly induced VCAM-1 and ICAM-1 expression on HMVEC-LBl. Although DEX showed no effect on the TNF-a-induced ICAM-1, TNF-a-induced VCAM-1 was enhanced by DEX treatment. TNF-a markedly induced CXCL1, CXCL8, CXCL10 and CCL5 in both HMVEC-LBl and BSMC. All of the TNF-a-induced chemokines in BSMC were significantly attenuated by DEX, whereas TNF-a-induced CXCL1 and CXCL8 in HMVEC-LBl were not affected at all. TNF-ainduced CXCL10 and CCL5 in HMVEC-LBl were enhanced by DEX. Furthermore, TNF-a-induced apoptosis of HMVEC-LBl was blocked by DEX. CONCLUSIONS: Some unfavorable effects of corticosteroid were found in TNF-a-mediated in vitro reactions by HMVEC-LBl but not by BSMC, suggesting that extravasation of leukocytes and survival of endothelial cells may be upregulated in vivo. Corticosteroids should be given with
J ALLERGY CLIN IMMUNOL FEBRUARY 2008
caution to severe asthmatic patients with a marked increase in TNF-a production. Funding: National Institute of Biomedical Innovation
785
Quercetin Potentially Ameliorates the C5a-induced Acute Peritonitis J. J. Aarthi1, S. D. Kumar2, P. N. Pushparaj1; 1Department of Physiology, National University of Singapore, Singapore, SINGAPORE, 2Department of Anatomy, National University of Singapore, Singapore, SINGAPORE. RATIONALE: To investigate the effects of Quercetin in the anaphylatoxin C5a-triggered responses in vivo. Complement cascade activation leads to the generation of the potent proinflammatory anaphylatoxin, C5a. Significant amounts of C5a, as well as other complement products in the blood, can lead to a series of adverse effects associated with a variety of pathologies. METHODS: Acute inflammation in the peritoneal cavity was induced by an intraperitoneal (i.p.) injection of recombinant human C5a (2 mg per mouse in a final volume of 100 ml). Quercetin (75 mM/100 ml of PBS) was i.v. injected 10 min before C5a i.p. injection. Four mice were used for each group per experiment, and the experiments were conducted three times. The i.v. administration of C5a triggers a rapid reduction in neutrophil level but pre-treating mice with the Quercetin 10 min before the C5a i.v. administration substantially inhibited the neutropenia and increase in the serum levels of IL-6 and TNF-a. The C5a i.p. injection triggered a fast recruitment of neutrophils followed by monocytes into the peritoneal cavity. Extravasation due to the vascular permeability was also observed: RESULTS: In mice pretreated with Quercetin, there was a significant reduction on the C5a-triggered neutrophil and monocyte infiltration, and a marked reduction on the Evans blue influx. Immunohistochemical analysis had shown that the expression of cell adhesion molecules (CAMs) such as P-Selecin, ICAM-1, and VCAM-1 were significantly down-regulated in the peritoneal membrane of the Quercetin-treated mice. CONCLUSIONS: These observations suggest that Quercetin treatment potentially blocks the C5a-triggered inflammatory responses in vivo. Funding: National University of Singapore
786
Micro RNA-Directed Translation Repression Is DownRegulated in Activated Bronchial Epithelial Cells Y. Zhai; UT Medical School, Houston, TX. RATIONALE: Since activation of bronchial epithelial cells causes elevation of the levels of inflammation mediators and tissue-remodeling factors, we examined the role of micro RNA (miRNA)-directed translation repression in this regulation. METHODS: MicroRNAs in bronchial epithelial cells (BEAS-2B) were identified by microarray profiling. The effect of a representative miRNA on the protein synthesis of its target reporter mRNA was tested in BEAS-2B cells with or without TNF-a/IL-4 stimulation. RESULTS: Protein synthesis rate of the reporter mRNA carrying the miRNA target sites is higher in the cells treated with TNF-a/IL-4 than that in the non-treated cells. CONCLUSION: Allergic immune response in bronchial epithelial cells elicits down-regulation of miRNA-directed translation repression. Funding: Sandler