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Primary Immunodeficiencies in Patients with Recurrent Ear and Sinus Infections
AB68 Abstracts
244
SUNDAY
Immune Complex-Mediated Damage Is Under Complement Pathway Control Eveline Y. Wu, MD, Garren Hester, BS, Haixiang Jiang, MD, PhD, Michael Frank, MD, FAAAAI; Duke University Medical Center. RATIONALE: IgG receptors (FcgR) are critical in immune complex (IC)-mediated tissue damage. We performed reverse passive Arthus reactions, a model of IC-mediated cutaneous vasculitis, in normal C57Bl/6 mice and mice genetically deficient in C1q, C4, C3, and Factor B. We hypothesized classical complement pathway (CP) down-regulates while alternative complement pathway (AP) promotes IC binding to activating FcgR; therefore, CP deficiency (C1q-/-, C4-/-, C3-/-) would enhance the IC-induced cutaneous vasculitis, while AP deficiency (FB-/-) would diminish it. METHODS: Sedated, shaved mice were injected intradermally with 20ml PBS on one side and rabbit anti-BSA IgG 5mg on the opposite. BSA 100mg and 125Iodine-labeled BSA 1.25mg with 1% Evans blue was then injected intravenously. After 4 hours, skin sections were weighed, radioactivity was measured (cpm/gm of tissue), and an Arthus index (AI) was calculated for each mouse (cpm/gm treated skin/control skin cpm/gm). Significant differences in AIs were analyzed by Kruskal-Wallis test, followed by MannWhitney tests. RESULTS: Complement absence strongly influenced degree of cutaneous vasculitis (p<0.001). Compared to normal mice (mean 2.561.2), C1q-/(mean 3.660.8, p<0.05) and C4-/- (mean 4.762.3, p<0.05) exhibited more extensive vasculitis, and C3-/- (mean 3.560.5, p50.05) trended towards greater vasculitis. FB-/- (mean 1.460.4, p<0.05) mice exhibited reduced vasculitis. CONCLUSIONS: Cutaneous vasculitis is significantly greater in CPdeficient mice and reduced in AP-deficient mice. Our results are first to provide in vivo evidence that complement critically influences IC and FcgR-mediated inflammation. They also refute previous data suggesting complement possesses no role in Arthus-induced vasculitis and may further understanding of why CP-deficient individuals are susceptible to autoimmunity.
245
Simultaneous Detection of Total and Allergen-Specific IgE in Human Plasma Using Multiplex Array Technology Meredith Lohman, MS, Martin D. Chapman, PhD, FAAAAI, Eva King, MSc, PhD; Indoor Biotechnologies, Inc., Charlottesville, VA. RATIONALE: Methods used for quantitative measurement of IgE in human serum or plasma analyze one analyte per test (FEIA). Chip-based multiplex technology (ISAC) provides semi-quantitative results for multiple analytes. The aim of this study was to develop a quantitative multiplex array for total and specific IgE that simultaneously detects multiple analytes, and validate the method by comparing results with streptavidinImmunoCAP. METHODS: IgE concentration in human plasma was detected using fluorescent microspheres covalently coupled to antibodies that were saturated with purified allergens. Human plasma samples (n579) were analyzed by 12-plex array and streptavidin-ImmunoCAP to determine total IgE as well as IgE specific to Der p 1, Der f 1, Der p 2, Der f 2, Fel d 1, Can f 1, Mus m 1, Rat n 1, Bet v 1, Phl p 5, and Alt a 1. RESULTS: IgE measurements by 12-plex array were reproducible both within and between assays: mean coefficients of variance (CV%) were 16.1% for intra-assay triplicates, and 17.1% for triplicate inter-assay analysis. The array demonstrated good parallelism between serial dilutions within the dynamic range (CV516.4%). Results obtained by multiplex array versus streptavidin-ImmunoCAP correlated closely (mean r50.88, p<0.001), with a mean CV of 56.5%, and a concordance of 84.2%. The total sample volume required for the 12-plex test was 30ml compared to 800ml required for ImmunoCAP. CONCLUSIONS: The multiplex array provides reproducible quantitative IgE measurements in human plasma, while using only a fraction of the sample volume required in other methods.
J ALLERGY CLIN IMMUNOL FEBRUARY 2013
246
Primary Immunodeficiencies in Patients with Recurrent Ear and Sinus Infections Chadi Makary, MD1, Rafka Chaiban, MD2, Benjamin Addicks2, Yesim Yilmaz Demirdag, MD2; 1West Virginina University School of Medicine, Morgantown, WV, 2West Virginia University School of Medicine, Morgantown, WV. RATIONALE: Primary immunodeficiencies (PIDs) frequently present with otolaryngologic infections but they are usually overlooked mostly because upper respiratory tract infections are common in general population. We aimed to determine the frequency of PIDs in patients with recurrent and/or chronic ENT infections. METHODS: We have started reviewing records of patients referred to Allergy/Immunology clinic with the diagnosis of chronic rhinitis, rhinosinusitis, recurrent sinusitis, recurrent otitis, and chronic otitis between July 2009 and July 2012. Patients who met the criteria of chronic (>12 weeks) or recurrent (>3 times/year) sinusitis and/or ear infections underwent immunologic evaluation. Patients with cystic fibrosis and anatomic defects were excluded. Demographic data, and associated factors, including recurrent pneumonias, asthma, and smoking exposure, were reviewed. Immunologic evaluation included CBC with differential, IgG, IgM, and IgA levels, lymphocyte subsets by flow cytometry and complement classic pathway assay (CH50) as well as baseline and post-vaccination pneumococcal and tetanus antibody titers were reviewed. RESULTS: There were 185 patients referred and to date, we have reviewed 82 charts. Forty-three patients (33 children, 10 adults) underwent immunologic evaluation. Eight patients (18.6%) had abnormal immunologic findings: 4 had low IgG, 2 had selective IgA deficiency, and 2 patients had specific antibody deficiency. Twenty-five percent (2/8) of these patients did not have any history of lower respiratory tract infections. CONCLUSIONS: Primary immunodeficiency disorders are not rare in patients with recurrent and/or chronic ENT infections and they should be considered in the differential diagnosis.
244
SUNDAY
Immune Complex-Mediated Damage Is Under Complement Pathway Control Eveline Y. Wu, MD, Garren Hester, BS, Haixiang Jiang, MD, PhD, Michael Frank, MD, FAAAAI; Duke University Medical Center. RATIONALE: IgG receptors (FcgR) are critical in immune complex (IC)-mediated tissue damage. We performed reverse passive Arthus reactions, a model of IC-mediated cutaneous vasculitis, in normal C57Bl/6 mice and mice genetically deficient in C1q, C4, C3, and Factor B. We hypothesized classical complement pathway (CP) down-regulates while alternative complement pathway (AP) promotes IC binding to activating FcgR; therefore, CP deficiency (C1q-/-, C4-/-, C3-/-) would enhance the IC-induced cutaneous vasculitis, while AP deficiency (FB-/-) would diminish it. METHODS: Sedated, shaved mice were injected intradermally with 20ml PBS on one side and rabbit anti-BSA IgG 5mg on the opposite. BSA 100mg and 125Iodine-labeled BSA 1.25mg with 1% Evans blue was then injected intravenously. After 4 hours, skin sections were weighed, radioactivity was measured (cpm/gm of tissue), and an Arthus index (AI) was calculated for each mouse (cpm/gm treated skin/control skin cpm/gm). Significant differences in AIs were analyzed by Kruskal-Wallis test, followed by MannWhitney tests. RESULTS: Complement absence strongly influenced degree of cutaneous vasculitis (p<0.001). Compared to normal mice (mean 2.561.2), C1q-/(mean 3.660.8, p<0.05) and C4-/- (mean 4.762.3, p<0.05) exhibited more extensive vasculitis, and C3-/- (mean 3.560.5, p50.05) trended towards greater vasculitis. FB-/- (mean 1.460.4, p<0.05) mice exhibited reduced vasculitis. CONCLUSIONS: Cutaneous vasculitis is significantly greater in CPdeficient mice and reduced in AP-deficient mice. Our results are first to provide in vivo evidence that complement critically influences IC and FcgR-mediated inflammation. They also refute previous data suggesting complement possesses no role in Arthus-induced vasculitis and may further understanding of why CP-deficient individuals are susceptible to autoimmunity.
245
Simultaneous Detection of Total and Allergen-Specific IgE in Human Plasma Using Multiplex Array Technology Meredith Lohman, MS, Martin D. Chapman, PhD, FAAAAI, Eva King, MSc, PhD; Indoor Biotechnologies, Inc., Charlottesville, VA. RATIONALE: Methods used for quantitative measurement of IgE in human serum or plasma analyze one analyte per test (FEIA). Chip-based multiplex technology (ISAC) provides semi-quantitative results for multiple analytes. The aim of this study was to develop a quantitative multiplex array for total and specific IgE that simultaneously detects multiple analytes, and validate the method by comparing results with streptavidinImmunoCAP. METHODS: IgE concentration in human plasma was detected using fluorescent microspheres covalently coupled to antibodies that were saturated with purified allergens. Human plasma samples (n579) were analyzed by 12-plex array and streptavidin-ImmunoCAP to determine total IgE as well as IgE specific to Der p 1, Der f 1, Der p 2, Der f 2, Fel d 1, Can f 1, Mus m 1, Rat n 1, Bet v 1, Phl p 5, and Alt a 1. RESULTS: IgE measurements by 12-plex array were reproducible both within and between assays: mean coefficients of variance (CV%) were 16.1% for intra-assay triplicates, and 17.1% for triplicate inter-assay analysis. The array demonstrated good parallelism between serial dilutions within the dynamic range (CV516.4%). Results obtained by multiplex array versus streptavidin-ImmunoCAP correlated closely (mean r50.88, p<0.001), with a mean CV of 56.5%, and a concordance of 84.2%. The total sample volume required for the 12-plex test was 30ml compared to 800ml required for ImmunoCAP. CONCLUSIONS: The multiplex array provides reproducible quantitative IgE measurements in human plasma, while using only a fraction of the sample volume required in other methods.
J ALLERGY CLIN IMMUNOL FEBRUARY 2013
246
Primary Immunodeficiencies in Patients with Recurrent Ear and Sinus Infections Chadi Makary, MD1, Rafka Chaiban, MD2, Benjamin Addicks2, Yesim Yilmaz Demirdag, MD2; 1West Virginina University School of Medicine, Morgantown, WV, 2West Virginia University School of Medicine, Morgantown, WV. RATIONALE: Primary immunodeficiencies (PIDs) frequently present with otolaryngologic infections but they are usually overlooked mostly because upper respiratory tract infections are common in general population. We aimed to determine the frequency of PIDs in patients with recurrent and/or chronic ENT infections. METHODS: We have started reviewing records of patients referred to Allergy/Immunology clinic with the diagnosis of chronic rhinitis, rhinosinusitis, recurrent sinusitis, recurrent otitis, and chronic otitis between July 2009 and July 2012. Patients who met the criteria of chronic (>12 weeks) or recurrent (>3 times/year) sinusitis and/or ear infections underwent immunologic evaluation. Patients with cystic fibrosis and anatomic defects were excluded. Demographic data, and associated factors, including recurrent pneumonias, asthma, and smoking exposure, were reviewed. Immunologic evaluation included CBC with differential, IgG, IgM, and IgA levels, lymphocyte subsets by flow cytometry and complement classic pathway assay (CH50) as well as baseline and post-vaccination pneumococcal and tetanus antibody titers were reviewed. RESULTS: There were 185 patients referred and to date, we have reviewed 82 charts. Forty-three patients (33 children, 10 adults) underwent immunologic evaluation. Eight patients (18.6%) had abnormal immunologic findings: 4 had low IgG, 2 had selective IgA deficiency, and 2 patients had specific antibody deficiency. Twenty-five percent (2/8) of these patients did not have any history of lower respiratory tract infections. CONCLUSIONS: Primary immunodeficiency disorders are not rare in patients with recurrent and/or chronic ENT infections and they should be considered in the differential diagnosis.