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Congenital NEMO Alteration at Position 223 causes Ectodermal Dysplasia and Immunodeficiency with Normogammaglobulinemia
Abstracts AB77
J ALLERGY CLIN IMMUNOL VOLUME 125, NUMBER 2
Congenital NEMO Alteration at Position 223 causes Ectodermal Dysplasia and Immunodeficiency with Normogammaglobulinemia G. Karamchandani-Patel1, R. U. Sorensen1, E. Hanson2, R. Saltzman2, J. Orange2; 1Louisiana State University Health Sciences Center, New Orleans, LA, 2University of Pennsylvania School of Medicine, Philadelphia, PA. RATIONALE: Hypomorphic mutations in the NEMO gene result in a variable syndrome of somatic and immunological abnormalities. It is becoming increasingly apparent that there are clinically relevant genotype-phenotype associations in this disease. We studied two unrelated boys with novel NEMO mutations resulting in alterations at position 223 to determine if they had similar phenotypes. METHODS: Clinical and immunologic characteristics as well as NF-kB activation were defined in two patients with novel NEMO mutations predicted to affect residue 223. Since this specific region of the NEMO protein has not been previously affected, consideration was given to a new genotype-phenotype correlation. RESULTS: Both patients were diagnosed at age two with hypohydrotic ectodermal dysplasia. Immunoglobulin levels, lymphocyte subpopulations and lymphocyte proliferation studies were normal. Patient 1 had recurrent sinopulmonary infections and pneumococcal antibody titers protective to only 1/14 serotypes. Patient 2 had a necrotizing soft tissue MRSA infection at 16 months and streptococcal angiosus bacteremia with subdural empyema at 26 months. DNA sequencing of the NEMO gene in both patients revealed mutations in exon 5 corresponding to position 223. Patient 1 had a 3 nucleotide deletion, c.667_669delGAG predicting the deletion of glutamine 223. Patient 2 had a missense mutation resulting in a nucleotide 667G>A which predicts Glu223>Lys substitution. Infectious susceptibility in both patients improved after immunoglobulin replacement therapy. CONCLUSIONS: Two different novel mutations affecting NEMO position 223 resulted in clinically relevant and similar phenotypes. This underscores the consideration of genotype-phenotype correlations in this disease and defines residue 223 as dispensable for quantitative, but not qualitative immunoglobulin production.
303
A Comparison Of The Effects Of TNF, IL-12/IL-23p40, And TGFb1 Peptide-based Vaccines In The Amelioration Of TNBSinduced Chronic Colitis q. guan1,2, Y. Ma2, C. Hillman2, A. G. Ma1,2, G. Zhou2, G. Qin3, C. R. Weiss1,2, Z. Peng1,2; 1Department of Immunology, University of Manitoba, Winnipeg, MB, CANADA, 2Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, CANADA, 3Department of Pathology, University of Manitoba, Winnipeg, MB, CANADA. RATIONALE: TNF, IL-12, IL-23 and TGFb1 are key cytokines in the pathogenesis of Crohn’s disease. We previously developed three mouse cytokine peptide-based virus-like particle vaccines against TNF, the p40 subunit shared by IL-12 and IL-23, and TGFb1, respectively. These vaccines elicited specific antibodies to the target cytokine and their antisera inhibited the bioactivity of the target. Here, we evaluated the effects of the vaccines separately and paralelly in murine chronic colitis, induced by intrarectal administrations of trinitrobenzene sulfonic acid (TNBS). METHODS: Mice were subcutaneously immunized with vaccine, vaccine carrier or saline three times and then intra-rectally administered TNBS weekly for 7 weeks. Serum cytokine-specific IgG, body weight, histology, collagen, and cytokines in colon tissue, and Th1 and Th17 cells in mesenteric lymph nodes (MLN) were measured. RESULTS: Vaccines induced high and long-lasting specific IgG antibodies to target cytokine. Vaccinated mice had less body weight loss and a decrease of inflammation, collagen deposition, and expression of p40, IL-23 and IL-17 proteins in colon tissues, compared with carrier and saline groups. Among them, p40 vaccine showed relatively stronger effects in body weight gain and colon inflammation than the other 2 vaccines. In p40 vaccinated mice, more parameters were analyzed. p40 vaccine decreased mRNA expression of p40, IL-10 and Bcl-2 and down-regulated
the percentages of Th1 and Th17 cells in MLN. CONCLUSIONS: Targeting TNF or IL-12/IL-23 or TGFb1, especially IL-12/IL-23, by employing a vaccine is effective in the amelioration of intestinal inflammation and fibrosis, providing a novel approach in the long-term treatment of Crohn’s disease.
304
Human Papillomavirus (HPV)-Specific T-cells Recognizing Dominant E2/E6 Epitopes Elicit Reduced IFN-g in Patients with Recurrent Respiratory Papillomatosis (RRP) E. James1, J. A. DeVoti2,3, D. W. Rosenthal3,4, L. J. Hatam3, B. M. Steinberg2,5, A. Abramson5, W. W. Kwok1, V. R. Bonagura3,4; 1Benaroya Research Institute, Seattle, WA, 2Feinstein Institute for Medical Research, North Shore-LIJ Health System, Manhasset, NY, 3Division of Allergy/ Immunology, North Shore LIJ Health System, Manhasset, NY, 4Elmezzi Graduate School of Molecular Medicine, North Shore LIJ Health System, Manhasset, NY, 5Department of Otolaryngology, Long Island Jewish Medical Center, New Hyde Park, NY. RATIONALE: RRP is an HPV-related disease, in which HLADRB1*0102 (DR0102) and DRB1*0301 (DR0301) haplotypes are associated with increased severity. TH1-like responses to E6/E2 proteins have been shown to protect against papillomavirus-induced disease in animals. Therefore, dysfunctional TH1 responses may be related to RPP disease severity in DR0102 and DR0301 subjects. METHODS: Using tetramer guided epitope mapping, we identified immunogenic peptides within HPV-11 early proteins (E6/E2) restricted by DR0102 and/or DR0301. Peptide binding, tetramer staining, and proliferation assays identified minimal epitopes within these peptides. The cytokine profile of sorted tetramer positive T-cell lines from RPP patients (n56) and controls (n510) was measured using a cytokine capture assay, to determine E2/E6-responsive T-cell polarization. RESULTS: Two distinct E6/E2 peptides (E6113-132 and E21-20) contained DR0102 and DR0301 restricted epitopes respectively. An additional peptide (E2281-300) contained an epitope presented by both DR0102 and DR0301. Minimal epitopes within these peptides bound to recombinant DR protein, gave positive tetramer staining for sorted T-cell lines, and elicited T-cell proliferation in both RPP patients and HLA-matched healthy controls. While the magnitude of responses to these epitopes was similar in both groups, IFN-g secretion was substantially lower in T-cell lines isolated from RPP patients. CONCLUSIONS: CD4+ T-cells specific for E2/E6 epitopes are easily detected in RPP patients and healthy controls with DR0102 and DR0301 haplotypes. However, RPP patients exhibit HPV-specific, immune dysregulation, indicated by a lack of IFN-g secretion. Therefore, therapeutic vaccination or other interventions that repolarize T-cell responses and restore TH1-like cytokine responses to HPV proteins could improve outcomes for RPP patients.
SUNDAY
302
J ALLERGY CLIN IMMUNOL VOLUME 125, NUMBER 2
Congenital NEMO Alteration at Position 223 causes Ectodermal Dysplasia and Immunodeficiency with Normogammaglobulinemia G. Karamchandani-Patel1, R. U. Sorensen1, E. Hanson2, R. Saltzman2, J. Orange2; 1Louisiana State University Health Sciences Center, New Orleans, LA, 2University of Pennsylvania School of Medicine, Philadelphia, PA. RATIONALE: Hypomorphic mutations in the NEMO gene result in a variable syndrome of somatic and immunological abnormalities. It is becoming increasingly apparent that there are clinically relevant genotype-phenotype associations in this disease. We studied two unrelated boys with novel NEMO mutations resulting in alterations at position 223 to determine if they had similar phenotypes. METHODS: Clinical and immunologic characteristics as well as NF-kB activation were defined in two patients with novel NEMO mutations predicted to affect residue 223. Since this specific region of the NEMO protein has not been previously affected, consideration was given to a new genotype-phenotype correlation. RESULTS: Both patients were diagnosed at age two with hypohydrotic ectodermal dysplasia. Immunoglobulin levels, lymphocyte subpopulations and lymphocyte proliferation studies were normal. Patient 1 had recurrent sinopulmonary infections and pneumococcal antibody titers protective to only 1/14 serotypes. Patient 2 had a necrotizing soft tissue MRSA infection at 16 months and streptococcal angiosus bacteremia with subdural empyema at 26 months. DNA sequencing of the NEMO gene in both patients revealed mutations in exon 5 corresponding to position 223. Patient 1 had a 3 nucleotide deletion, c.667_669delGAG predicting the deletion of glutamine 223. Patient 2 had a missense mutation resulting in a nucleotide 667G>A which predicts Glu223>Lys substitution. Infectious susceptibility in both patients improved after immunoglobulin replacement therapy. CONCLUSIONS: Two different novel mutations affecting NEMO position 223 resulted in clinically relevant and similar phenotypes. This underscores the consideration of genotype-phenotype correlations in this disease and defines residue 223 as dispensable for quantitative, but not qualitative immunoglobulin production.
303
A Comparison Of The Effects Of TNF, IL-12/IL-23p40, And TGFb1 Peptide-based Vaccines In The Amelioration Of TNBSinduced Chronic Colitis q. guan1,2, Y. Ma2, C. Hillman2, A. G. Ma1,2, G. Zhou2, G. Qin3, C. R. Weiss1,2, Z. Peng1,2; 1Department of Immunology, University of Manitoba, Winnipeg, MB, CANADA, 2Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, CANADA, 3Department of Pathology, University of Manitoba, Winnipeg, MB, CANADA. RATIONALE: TNF, IL-12, IL-23 and TGFb1 are key cytokines in the pathogenesis of Crohn’s disease. We previously developed three mouse cytokine peptide-based virus-like particle vaccines against TNF, the p40 subunit shared by IL-12 and IL-23, and TGFb1, respectively. These vaccines elicited specific antibodies to the target cytokine and their antisera inhibited the bioactivity of the target. Here, we evaluated the effects of the vaccines separately and paralelly in murine chronic colitis, induced by intrarectal administrations of trinitrobenzene sulfonic acid (TNBS). METHODS: Mice were subcutaneously immunized with vaccine, vaccine carrier or saline three times and then intra-rectally administered TNBS weekly for 7 weeks. Serum cytokine-specific IgG, body weight, histology, collagen, and cytokines in colon tissue, and Th1 and Th17 cells in mesenteric lymph nodes (MLN) were measured. RESULTS: Vaccines induced high and long-lasting specific IgG antibodies to target cytokine. Vaccinated mice had less body weight loss and a decrease of inflammation, collagen deposition, and expression of p40, IL-23 and IL-17 proteins in colon tissues, compared with carrier and saline groups. Among them, p40 vaccine showed relatively stronger effects in body weight gain and colon inflammation than the other 2 vaccines. In p40 vaccinated mice, more parameters were analyzed. p40 vaccine decreased mRNA expression of p40, IL-10 and Bcl-2 and down-regulated
the percentages of Th1 and Th17 cells in MLN. CONCLUSIONS: Targeting TNF or IL-12/IL-23 or TGFb1, especially IL-12/IL-23, by employing a vaccine is effective in the amelioration of intestinal inflammation and fibrosis, providing a novel approach in the long-term treatment of Crohn’s disease.
304
Human Papillomavirus (HPV)-Specific T-cells Recognizing Dominant E2/E6 Epitopes Elicit Reduced IFN-g in Patients with Recurrent Respiratory Papillomatosis (RRP) E. James1, J. A. DeVoti2,3, D. W. Rosenthal3,4, L. J. Hatam3, B. M. Steinberg2,5, A. Abramson5, W. W. Kwok1, V. R. Bonagura3,4; 1Benaroya Research Institute, Seattle, WA, 2Feinstein Institute for Medical Research, North Shore-LIJ Health System, Manhasset, NY, 3Division of Allergy/ Immunology, North Shore LIJ Health System, Manhasset, NY, 4Elmezzi Graduate School of Molecular Medicine, North Shore LIJ Health System, Manhasset, NY, 5Department of Otolaryngology, Long Island Jewish Medical Center, New Hyde Park, NY. RATIONALE: RRP is an HPV-related disease, in which HLADRB1*0102 (DR0102) and DRB1*0301 (DR0301) haplotypes are associated with increased severity. TH1-like responses to E6/E2 proteins have been shown to protect against papillomavirus-induced disease in animals. Therefore, dysfunctional TH1 responses may be related to RPP disease severity in DR0102 and DR0301 subjects. METHODS: Using tetramer guided epitope mapping, we identified immunogenic peptides within HPV-11 early proteins (E6/E2) restricted by DR0102 and/or DR0301. Peptide binding, tetramer staining, and proliferation assays identified minimal epitopes within these peptides. The cytokine profile of sorted tetramer positive T-cell lines from RPP patients (n56) and controls (n510) was measured using a cytokine capture assay, to determine E2/E6-responsive T-cell polarization. RESULTS: Two distinct E6/E2 peptides (E6113-132 and E21-20) contained DR0102 and DR0301 restricted epitopes respectively. An additional peptide (E2281-300) contained an epitope presented by both DR0102 and DR0301. Minimal epitopes within these peptides bound to recombinant DR protein, gave positive tetramer staining for sorted T-cell lines, and elicited T-cell proliferation in both RPP patients and HLA-matched healthy controls. While the magnitude of responses to these epitopes was similar in both groups, IFN-g secretion was substantially lower in T-cell lines isolated from RPP patients. CONCLUSIONS: CD4+ T-cells specific for E2/E6 epitopes are easily detected in RPP patients and healthy controls with DR0102 and DR0301 haplotypes. However, RPP patients exhibit HPV-specific, immune dysregulation, indicated by a lack of IFN-g secretion. Therefore, therapeutic vaccination or other interventions that repolarize T-cell responses and restore TH1-like cytokine responses to HPV proteins could improve outcomes for RPP patients.
SUNDAY
302