4 Receptor Antagonist on Chemokine and Cytokine Synthesis By PBMC and Dendritic Cells Derived from PBMC

4 Receptor Antagonist on Chemokine and Cytokine Synthesis By PBMC and Dendritic Cells Derived from PBMC

Abstracts AB119 J ALLERGY CLIN IMMUNOL VOLUME 137, NUMBER 2 Impact of an H3/4 Receptor Antagonist on Chemokine and Cytokine Synthesis By PBMC and De...

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Abstracts AB119

J ALLERGY CLIN IMMUNOL VOLUME 137, NUMBER 2

Impact of an H3/4 Receptor Antagonist on Chemokine and Cytokine Synthesis By PBMC and Dendritic Cells Derived from PBMC

Roman Khanferyan, MD, PhD1, V. Evstratova1, N. Rieger1, Lawrence M. DuBuske, MD, FAAAAI2,3; 1Institute of Nutrition, Moscow, Russia, 2 Immunology Research Institute of New England, Gardner, MA, 3George Washington University School of Medicine, Washington, DC. RATIONALE: Histamine H3 and H4 histamine receptors are involved in immune modulation. Peripheral blood mononuclear cells (PBMC) and dendritic cells (DCs) derived from PBMC synthesize cytokines and chemokines which may be modulated by blockade of H3/ H4 histamine receptors. METHODS: PBMC and DCs derived from 10 healthy donors were cultivated in the presence of dual H3/4 histamine receptor antagonist Ciproxifan (10-5M). The concentrations of cytokines and chemokines in 48-hour cultures were assessed by Multiplex assays using Luminex xMAP technology. RESULTS: Cultivation of PBMC with the H3/4 antagonist significantly increased secretion of IL-4, IL-13, IL-18, IL-27, IL1a and IP-10 (p<0.05) and inhibited secretion of multiple chemo- and cytokines, SCF, GM-CSF, LIF, IL-2, -5, -6, -7, -9, -15, -31. In DC culture Ciproxifan significantly up-regulate the secretion of SCF, GM-CSF, IL1-a and IL-5 and down-regulates the production of IL-2, IL-6, IL-15 and LIF (p<0.05). Ciproxifan increases secretion of chemokines by PBMC (Eotaxin, RANTES, MIP-1a, MIP-1b) as well as DC (MCP-1 and RANTES). CONCLUSIONS: Histamine H3/4 receptors are involved in the regulation of secretion of cytokines and chemokines by both PBMC and DC derived from PBMC.

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Expression Pattern of Peripheral Blood Mononuclear Leucocyte GABA Receptors and Calcium Signaling Genes

Leonid P. Titov, MD, PhD1, A. B. Kapitau1, K. I. Pavlov1, I. M. Goloenko1, Lawrence M. DuBuske, MD, FAAAAI2,3; 1Republican Research and Practical Center for Epidemiology and Microbiology, Minsk, Belarus, 2 Immunology Research Institute of New England, Gardner, MA, 3George Washington University School of Medicine, Washington, DC. RATIONALE: Neuromediators including gamma-aminobutyric acid (GABA) have immune modulation, impacting airway epithelium of allergic asthma stimulating mucus production. METHODS: PBML gene expression profiling was performed using Human Discover Chipsä and Neurotransmitter Receptors and Regulators Arrayit Pathwaysä Focused microarrays (Arrayit corporation, California, USA). GABA A2 receptor polymorphic locus rs279826 analysis was performed using standard PCR with subsequent Mnl1 endonuclease restriction. RESULTS: Microarray methodology demonstrated significant expression levels in PBML for calcium-associated ion channels and second messengers: chloride channel 1, calcium activated gene, calmodulin 1 gene, S100 calcium-binding protein A7 gene, vacuolar ATFase gene (increasing level of expression: CLCA1 < CALM1 < S100A7 < VPS33B) with slight expression of cholinergic receptors and nicotinic genes compared with intensive expression of gamma-aminobutyric acid (GABA) A receptor alpha 2 gene. CONCLUSIONS: GABA A2 receptors have high expression in PBML. PBML GABA receptors and calcium associated second messenger expression indicate a significant peripheral effect of neurotransmitters.

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IL1-Beta Levels in Patients with Refractory Recurrent Pericarditis

Rushita Mehta, MD, Arye Rubinstein, MD; Albert Einstein College of Medicine and Montefiore Hospital, Bronx, NY. RATIONALE: Recurrent pericarditis is a debilitating condition thought to have an underlying immunologically-mediated mechanism. This condition is often resistant to conventional therapeutic options including nonsteroidal anti-inflammatory drugs, colchicine, and glucocorticoids. We report three cases of refractory recurrent pericarditis (RRP) who were found to have elevated serum IL1-b levels, one of whom responded to Anakinra, an IL1-receptor antagonist. These findings implicate IL1-b in RRP pathogenesis and suggest a therapeutic role for IL1-b blockage. METHODS: A retrospective chart review was conducted on 3 patients, aged 32-42 years, with confirmed RRP lasting 2-12 years. RESULTS: Case 1 is a 32 year old female with familial Mediterranean fever and a 12-year history of RRP unresponsive to steroids and colchicine. Serum IL1-b levels were elevated (69.6 pg/mL, normal <3.9), and treatment with IVIG and abortive Anakinra resulted in a clinical improvement. Case 2 is a 42 year-old female with hypogammaglobulinemia, lupus, Sjogren syndrome, and a three-year history of RRP despite treatment with glucocorticoids, colchicine, and toradol. IL1-b was markedly elevated (137.5 pg/mL). Case 3 is a 38 year-old male with chronic EBV infection and a three-year history of RRP unresponsive to colchicine and steroids. IL1-b was elevated (5 pg/mL). CONCLUSIONS: This case series describes three patients with recurrent refractory pericarditis who were found to have elevated IL1-b, one of whom responded to Anakinra, an IL-1-receptor antagonist. These cases indicate that IL1-b may be implicated in the pathogenesis of recurrent refractory pericarditis and suggest a potential role for Anakinra in the management of this disease.

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Rapid Molecular Identification and Quantification of Allergenic Pollen By Real-Time PCR

Michael Teng, PhD1, Mark C. Glaum, MD, PhD, FAAAAI2, Dennis K. Ledford, MD3; 1Division of Allergy and Immunology, Department of Internal Medicine, and the Joy McCann Culverhouse Airway Diseases Research Center, University of South Florida Morsani College of Medicine, Tampa, FL, 2Division of Allergy and Immunology, Department of Internal Medicine, Morsani College of Medicine, University of South Florida, Tampa, FL, 3University of South Florida and the James A. Haley VA Hospital, Tampa, FL. RATIONALE: Identifying and quantifying airborne pollen is important for causal assessment of allergic symptoms, for determining which pollens should be tested and for interpreting the results of allergy tests. Present pollen counting methods are time-consuming, require specific expertise and training, and are prone to subjective variability. Our goal is to devise a sensitive, high throughput method of quantifying pollen load in environmental samples. METHODS: 9 different pure pollen samples, including grass (Paspalum notatum), trees (Morella cerifera, Morus alba, Juniperus ashei, Pinus strobus, Quercus virginiana, Taxodium distichum, Ulmus americana), and ragweed (Ambrosia artemisifolia) were used for analysis (courtesy of Tom Greer PhD, Greer Laboratories). Genomic DNA (gDNA) was extracted from pure pollens using a modification of the DNAzol ES procedure. Primers specific for the 5.8S rRNA genes of the different pollens were designed for quantitative real-time PCR (qPCR) using a SYBRGreen-based PCR assay. RESULTS: Individual gDNA from the pollens was detectable to sub nanogram levels. Individual pollen gDNA was detected in samples consisting of a mixture of all 9 pollen gDNAs. Specific pollens were detected in gDNA extracted from randomly admixed samples of the various pollen samples. The selected primers provided accurate identification in mixed samples with variable efficiency. CONCLUSIONS: qPCR detection of the pollen gDNA is sensitive and specific. The current assay can be expanded to include additional species and/ or modified to detect pollens native to distinct geographical regions. Actual environmental sample quantification is necessary to prove applicability.

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