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hypogammaglobulinemia - Distinct Patterns Of Cytokine Production And Gene Expression Profiles
Abstracts AB231
J ALLERGY CLIN IMMUNOL VOLUME 127, NUMBER 2
Evidence of macrophage activation in Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal Infections (PANDAs) M. Longley, E. Kellner, S. Hudey, S. Lukas, P. Sriaroon, J. Sleasman, E. Perez, T. Murphy, M. Dorsey; University of South Florida/All Children’s Hospital, St. Petersburg, FL. RATIONALE: Children with PANDAS develop severe behavioral changes with obsessive compulsive disorder (OCD) or tic symptoms, associated with group A beta-hemolytic streptococcal infections (GAS). Cross reactive antibodies to oligodendrocytes is a proposed link between infection and this disorder. sCD14 is released after TLR2 and TLR4 ligation by bacterial peptidoglycan and lipopolysaccharide (LPS), respectively. N-acetylglucosamine, present in bacterial peptidoglycan is a dominant epitope of GAS and the neuronal cell surface molecule, GM1. Antibodies to N-acetylglucosamine have been linked to PANDAS related conditions. To investigate the role of macrophage activation in PANDAS we measured sCD14 as a surrogate marker of immune activation. METHODS: sCD14 ELISA was performed on samples from healthy subjects and prospectively followed PANDAS patients who met strict criteria after extensive neuropsychiatric evaluation. Statistical analysis was calculated with GraphPad Prism software using Mann-whitney U t-test and Spearman correlation. Demographic data was collected including clinical presentation, therapy, and presence of co-morbid conditions. Consent for all subjects was obtained using IRB approved protocol. RESULTS: sCD14 in PANDAS patients (n521) was increased significantly (mean 6359 ng/ml 6 1187) compared to healthy donors (n522) (mean 1777 ng/ml 6 1051; p <0.0001). sCD14 correlated positively with ASO titers (r50.5314; p50.0192). History of maternal autoimmune disease was present in six of twenty one patients with PANDAS (28.6%). CONCLUSIONS: Elevated sCD14 levels reflecting macrophage activation parallel previous findings. Correlation with increased ASO titers supports chronic immune activation via bacterial peptidoglycan. Together these findings strengthen the link between innate immune activation and other immunoglogical mechanisms involved in PANDAS.
Characterization Of Children With Autism Spectrum Disorders (asd) Requiring Intravenous Immunoglobulin (ivig) For Specific Polysaccharide Antibody Deficiency (spad)/ hypogammaglobulinemia - Distinct Patterns Of Cytokine Production And Gene Expression Profiles H. Jyonouchi, L. Geng, S. Kapoor, D. Streck, G. Toruner; UMDNJ-New Jersey Medical School, Newark, NJ. RATIONALE: The fact that ASD children are heterogeneous indicates a need for sub-grouping. This study evaluated whether ASD children, with defined immunodeficiency requiring IVIG treatment, have immune abnormalities that are distinct from ASD children not requiring IVIG treatment or non-ASD children treated with IVIG. METHODS: We assessed cytokine production patterns produced by peripheral blood (PB) mononuclear cells in ASD children treated with IVIG secondary to SPAD/ hypogammaglobulinemia (Test group, N510), control ASD (N549)/normal (N539) children, and non-ASD children with similar immunodeficiency (ID) requiring IVIG (N514) in addition to gene profiling of PB monocytes in 7/10 test group children and normal (N521) and ASD (N527) controls RESULTS: In the test group, we observed decreased production of proinflammatory cytokines (IL-6/IL-12/IL-23) and increased production of counter-regulatory cytokines (TGF-ß/sTNFRII) with TLR agonists as compared to normal controls (p<0.05). Also a decrease in production of IL-12 (with candida and PHA/ConA) and IL-17/IFN-g (with PHA) was observed (p<0.05). These changes were not observed in the control ASD and non-ASD/ID children. Up-regulation (>2-fold;1258 and 830 genes) and down-regulation (>2-fold;1235 and 653) in gene expression was observed in the test groups when compared with normal and ASD controls, respectively. In the test group, gene expression in TGF-ßR and NOTCH pathways were different from ASD controls (p<0.01). CONCLUSIONS: The ASD test group revealed distinct changes in cytokine production and mRNA expression. This difference is unlikely to be attributed to the ID dianosed in the test group, indicating a possible role of these changes in ASD pathogenesis in this subset of ASD.
892
894
Differences in Regulatory Capacity of Umbilical Cord Blood Mononuclear Cells Based on Maternal Atopic Risk K. M. Fraser, A. K. Ellis; Queen’s University, Kingston, ON, CANADA. RATIONALE: Maternal atopy is a known significant risk factor for the development of allergy in children, and may exert influence in utero on fetal immune cells. Atopy is generally viewed as a failure of immune tolerance, and we thus aimed to evaluate the effects of maternal atopy on umbilical cord blood (CB) mononuclear cell (MNC) production of regulatory cytokines following innate stimulation. METHODS: Pregnant women delivering via planned Caesarian Section at Kingston General Hospital were recruited for study participation, and maternal atopic status was recorded after written informed consent was obtained. CB MNCs were isolated from both atopic and non-atopic mothers (N54 each) following delivery, and incubated with 1mg/mL of lipopolysaccaride (LPS) or peptidoglycan (PGN), for 6 and 24 hours. Cell supernatants were collected and assayed for regulatory cytokine production by ELISA. RESULTS: Production of Interleukin-10 (IL-10) and Transforming Growth Factor-Beta (TGF-b) was significantly lower in the CB MNC from atopic mothers compared to non-atopic mothers following innate stimulation. Specifically, after 24hrs incubation with PGN, IL-10 levels were 1.3 fold higher in MNCs supernatants from non-atopic mothers compared to their atopic counterparts (p<0.05) The level of TGF-b secreted by the MNCs of non-atopic mothers following 6hr stimulation with LPS was 3.5 fold higher than those from allergic mothers (p<0.05). CONCLUSIONS: Cord blood MNCs from atopic mothers appear to have impaired regulatory capacity following innate stimulation compared to non-atopic controls, and may have relevance to the development of allergic disease in the offspring.
893
Levels of IgE in Acute Nephritis Patients as Related to the Efficacy of Treatment R. Khanferyan1, N. Fedicheva1, J. Shakhrai1, L. Gorbov1, L. M. DuBuske2; 1Kuban State Medical University, Krasnodar, RUSSIAN FEDERATION, 2Immunology Research Institute of New England, Gardner, MA. RATIONALE: Autoimmune glomerulonephritis is characterised by immune disturbances in the balance of Th1/Th2 cell activity with 20 to 40% of acute nephritis patients demonstrating chronic inflammation. This investigation evaluated the level of IgE synthesis in patients with acute nephritis and its dependence on efficacy of treatment. METHODS: Total IgE as well as IgE regulatory cytokines (IL4 and gIFN) were investigated in sera of 27 patients with acute nephritis by ELISA methods. Patients were divided into 2 groups: group A (15 patients) with high efficacy of treatment and group B (12) with low efficacy of treatment. RESULTS: Patients in both groups before treatment had similar IgE concentrations. After treatment the serum level of IgE in patients having high therapeutic efficacy decreased in 1.76 fold from 272.4 to 154.4 IU/ml while in patients with low therapeutic there was an increase in total IgE up to 2.8 fold. Alteration in total IgE production was highly dependent on the ratio of IL4 to IFN g. CONCLUSIONS: IgE may be involved in the mechanisms of acute and chronic immune inflammation in patients with nephritis and may be a prognostic indicator for the efficacy of treatment.
TUESDAY
891
J ALLERGY CLIN IMMUNOL VOLUME 127, NUMBER 2
Evidence of macrophage activation in Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal Infections (PANDAs) M. Longley, E. Kellner, S. Hudey, S. Lukas, P. Sriaroon, J. Sleasman, E. Perez, T. Murphy, M. Dorsey; University of South Florida/All Children’s Hospital, St. Petersburg, FL. RATIONALE: Children with PANDAS develop severe behavioral changes with obsessive compulsive disorder (OCD) or tic symptoms, associated with group A beta-hemolytic streptococcal infections (GAS). Cross reactive antibodies to oligodendrocytes is a proposed link between infection and this disorder. sCD14 is released after TLR2 and TLR4 ligation by bacterial peptidoglycan and lipopolysaccharide (LPS), respectively. N-acetylglucosamine, present in bacterial peptidoglycan is a dominant epitope of GAS and the neuronal cell surface molecule, GM1. Antibodies to N-acetylglucosamine have been linked to PANDAS related conditions. To investigate the role of macrophage activation in PANDAS we measured sCD14 as a surrogate marker of immune activation. METHODS: sCD14 ELISA was performed on samples from healthy subjects and prospectively followed PANDAS patients who met strict criteria after extensive neuropsychiatric evaluation. Statistical analysis was calculated with GraphPad Prism software using Mann-whitney U t-test and Spearman correlation. Demographic data was collected including clinical presentation, therapy, and presence of co-morbid conditions. Consent for all subjects was obtained using IRB approved protocol. RESULTS: sCD14 in PANDAS patients (n521) was increased significantly (mean 6359 ng/ml 6 1187) compared to healthy donors (n522) (mean 1777 ng/ml 6 1051; p <0.0001). sCD14 correlated positively with ASO titers (r50.5314; p50.0192). History of maternal autoimmune disease was present in six of twenty one patients with PANDAS (28.6%). CONCLUSIONS: Elevated sCD14 levels reflecting macrophage activation parallel previous findings. Correlation with increased ASO titers supports chronic immune activation via bacterial peptidoglycan. Together these findings strengthen the link between innate immune activation and other immunoglogical mechanisms involved in PANDAS.
Characterization Of Children With Autism Spectrum Disorders (asd) Requiring Intravenous Immunoglobulin (ivig) For Specific Polysaccharide Antibody Deficiency (spad)/ hypogammaglobulinemia - Distinct Patterns Of Cytokine Production And Gene Expression Profiles H. Jyonouchi, L. Geng, S. Kapoor, D. Streck, G. Toruner; UMDNJ-New Jersey Medical School, Newark, NJ. RATIONALE: The fact that ASD children are heterogeneous indicates a need for sub-grouping. This study evaluated whether ASD children, with defined immunodeficiency requiring IVIG treatment, have immune abnormalities that are distinct from ASD children not requiring IVIG treatment or non-ASD children treated with IVIG. METHODS: We assessed cytokine production patterns produced by peripheral blood (PB) mononuclear cells in ASD children treated with IVIG secondary to SPAD/ hypogammaglobulinemia (Test group, N510), control ASD (N549)/normal (N539) children, and non-ASD children with similar immunodeficiency (ID) requiring IVIG (N514) in addition to gene profiling of PB monocytes in 7/10 test group children and normal (N521) and ASD (N527) controls RESULTS: In the test group, we observed decreased production of proinflammatory cytokines (IL-6/IL-12/IL-23) and increased production of counter-regulatory cytokines (TGF-ß/sTNFRII) with TLR agonists as compared to normal controls (p<0.05). Also a decrease in production of IL-12 (with candida and PHA/ConA) and IL-17/IFN-g (with PHA) was observed (p<0.05). These changes were not observed in the control ASD and non-ASD/ID children. Up-regulation (>2-fold;1258 and 830 genes) and down-regulation (>2-fold;1235 and 653) in gene expression was observed in the test groups when compared with normal and ASD controls, respectively. In the test group, gene expression in TGF-ßR and NOTCH pathways were different from ASD controls (p<0.01). CONCLUSIONS: The ASD test group revealed distinct changes in cytokine production and mRNA expression. This difference is unlikely to be attributed to the ID dianosed in the test group, indicating a possible role of these changes in ASD pathogenesis in this subset of ASD.
892
894
Differences in Regulatory Capacity of Umbilical Cord Blood Mononuclear Cells Based on Maternal Atopic Risk K. M. Fraser, A. K. Ellis; Queen’s University, Kingston, ON, CANADA. RATIONALE: Maternal atopy is a known significant risk factor for the development of allergy in children, and may exert influence in utero on fetal immune cells. Atopy is generally viewed as a failure of immune tolerance, and we thus aimed to evaluate the effects of maternal atopy on umbilical cord blood (CB) mononuclear cell (MNC) production of regulatory cytokines following innate stimulation. METHODS: Pregnant women delivering via planned Caesarian Section at Kingston General Hospital were recruited for study participation, and maternal atopic status was recorded after written informed consent was obtained. CB MNCs were isolated from both atopic and non-atopic mothers (N54 each) following delivery, and incubated with 1mg/mL of lipopolysaccaride (LPS) or peptidoglycan (PGN), for 6 and 24 hours. Cell supernatants were collected and assayed for regulatory cytokine production by ELISA. RESULTS: Production of Interleukin-10 (IL-10) and Transforming Growth Factor-Beta (TGF-b) was significantly lower in the CB MNC from atopic mothers compared to non-atopic mothers following innate stimulation. Specifically, after 24hrs incubation with PGN, IL-10 levels were 1.3 fold higher in MNCs supernatants from non-atopic mothers compared to their atopic counterparts (p<0.05) The level of TGF-b secreted by the MNCs of non-atopic mothers following 6hr stimulation with LPS was 3.5 fold higher than those from allergic mothers (p<0.05). CONCLUSIONS: Cord blood MNCs from atopic mothers appear to have impaired regulatory capacity following innate stimulation compared to non-atopic controls, and may have relevance to the development of allergic disease in the offspring.
893
Levels of IgE in Acute Nephritis Patients as Related to the Efficacy of Treatment R. Khanferyan1, N. Fedicheva1, J. Shakhrai1, L. Gorbov1, L. M. DuBuske2; 1Kuban State Medical University, Krasnodar, RUSSIAN FEDERATION, 2Immunology Research Institute of New England, Gardner, MA. RATIONALE: Autoimmune glomerulonephritis is characterised by immune disturbances in the balance of Th1/Th2 cell activity with 20 to 40% of acute nephritis patients demonstrating chronic inflammation. This investigation evaluated the level of IgE synthesis in patients with acute nephritis and its dependence on efficacy of treatment. METHODS: Total IgE as well as IgE regulatory cytokines (IL4 and gIFN) were investigated in sera of 27 patients with acute nephritis by ELISA methods. Patients were divided into 2 groups: group A (15 patients) with high efficacy of treatment and group B (12) with low efficacy of treatment. RESULTS: Patients in both groups before treatment had similar IgE concentrations. After treatment the serum level of IgE in patients having high therapeutic efficacy decreased in 1.76 fold from 272.4 to 154.4 IU/ml while in patients with low therapeutic there was an increase in total IgE up to 2.8 fold. Alteration in total IgE production was highly dependent on the ratio of IL4 to IFN g. CONCLUSIONS: IgE may be involved in the mechanisms of acute and chronic immune inflammation in patients with nephritis and may be a prognostic indicator for the efficacy of treatment.
TUESDAY
891