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Immune response in tubulin induces hearing loss in mice
S16 Abstracts
J ALLERGY CLIN IMMUNOL FEBRUARY 2005
SATURDAY
Prevalence of Cystic Fibrosis in Children Who Present With Nasal Polyposis E. J. Schmitt, W. Neaville, T. Pongdee; Allergy/Immunology, University of Texas Southwestern Medical Center, Dallas, TX. RATIONALE: Children with cystic fibrosis have a higher incidence of nasal polyposis than unaffected children. Additionally, children with nasal polyposis are commonly screened for cystic fibrosis, whether or not other co-morbid features of cystic fibrosis are evident. The actual prevalence of cystic fibrosis diagnosed solely on the basis of isolated nasal polyposis disease has not been reported. METHODS: A medical records search identified all patients with an ICD-9 diagnosis code for cystic fibrosis and nasal polyps at Children’s Medical Center from 1999-2004. The order of each diagnosis was determined by chart review. RESULTS: We report 893 children with cystic fibrosis and 216 children with nasal polyps were seen at our institution over this five year period. Forty-five children were confirmed to have both diagnoses. Only 2 of 45 children developed nasal polyps as the initial manifestation that lead to the subsequent diagnosis of cystic fibrosis. Both of these subjects were otherwise healthy 6 year old Caucasian females, however one had chronic sinusitis with hypoplastic sinuses while the other was believed to have nasal allergies. We find a prevalence of 45/893 or 5% of cystic fibrosis patients with nasal polyps, but only 2/893 or 0.2% of the cystic fibrosis patients had their disease discovered because of nasal polyps. CONCLUSION: It is quite rare to diagnose cystic fibrosis secondary to the presence of nasal polyps. Furthermore, a suspicion for other etiologies aside from cystic fibrosis should be maintained in children who present with isolated nasal polyposis.
61
62
B Cell Superantigen-Induced Immune Complex Peritonitis
Anderson1,2,
Sporici1,
LaRosa2,
Levinson1,2; 1Philadel-
A. R. D. F. A. I. phia Veterans Administration Medical Center, Philadelphia, PA, 2University of Pennsylvania School of Medicine, Philadelphia, PA. RATIONALE: Staphylococcal protein A (SpA), a representative B cell superantigen (SAg), binds to the Fab regions of most VH3+ immunoglobulin molecules outside of their complementarity determining regions. We previously reported that the interaction of SpA with VH3+ immunoglobulin molecules produces a histologic pattern of inflammation in the skin of a rabbit indicative of immune complex injury. To begin to identify the cellular and molecular events contributing to this type of unconventional immune complex mediated inflammation, we established a mouse peritoneal Arthus model. METHODS: BALB/c mice were administered SpA intraperitoneally and either human polyclonal IgG (hIgG) or saline intravenously. Peritoneal lavage was performed 8 hours later and the fluid analyzed for differential white blood cell counts and levels of chemokines. VH3- depleted hIgG was substituted for hIgG in some experiments. Studies were also conducted in mast cell deficient and in TNF- knockout mice. RESULTS: BALB/c mice treated with SpA and hIgG developed a neutrophilic response that peaked eight hours after challenge. The response was dependent on the presence of VH3+ immunolgobulins. We also observed that mast cells and TNF- play obligatory roles in this reaction and that it is associated with the local release of the CXC chemokines MIP-2 and KC. CONCLUSIONS: These results provide further compelling evidence for the induction of immune-complex-mediated injury by a B cell SAg and
highlight important factors contributing to the pathogenesis of this novel type of inflammatory reaction. Funding: Veterans Administration Anti-Cyclic Citrullinated Peptide (CCP) Testing: Relevance in Rheumatoid Arthritis and Other Autoimmune Disorders J. M. Popov, M. Glovsky, K. Tokoro, O. Zhukov, F. Zarabpouri, K. Qu, A. Sferruzza; Immunology, Quest Diagnostics, San Juan Capistrano, CA. RATIONALE: CCP antibody is a relatively new, specific marker for detection of rheumatoid arthritis (RA) that can be used in conjunction with rheumatoid factor (RF) antibody. It is rarely present in other autoimmune diseases including systemic lupus erythematosus (SLE). In this study, we determined the frequency of CCP antibody in patients with RA and SLE as well as in patients with unknown or suspected autoimmune disease. METHODS: 516 sera sent to Quest Diagnostics for autoimmune testing (including ANA testing by EIA and IFA) and 113 sera from patients with RA (n=50) or SLE (n=63) were included. Presence of CCP antibody (IgG) was determined by enzyme-linked immunoassay. Further, presence of RF IgM was determined by latex agglutination. RESULTS: CCP antibody was present in 96% of RA cases and 3% of SLE cases. Thirty percent of the samples submitted for autoimmune testing (diagnosis unknown) were RF+/CCP+ and 70% were RF+/CCP-. Four of 120 (3%) ANA-positive samples were RF+/CCP+ and 27 (23%) were RF+/CCP-. CONCLUSIONS: The CCP antibody sensitivity in patients with RA is 96%. As expected, CCP antibody was rarely (3%) detected in patients with SLE. Patients with RF+/CCP- serotype are likely to have other diseases such as infection or immune complex diseases.
63
64
Immune Response in Tubulin Induces Hearing Loss in Mice
M. H. Kermany, Z. Bin, K. Mohammad, Z. Bin, J. Lee; Department of Medicine, University of Tennessee, Memphis, TN. RATIONALE: -Tubulin is an important molecule in the hair cells and supporting cells within the sensory epithelium and found to be an autoantigen in Méniere’s disease. Our goal is to induce hearing loss in mice with varying doses of -Tubulin antigen and evaluate the pathogenesis of autoimmune hearing loss induced by tubulin in mice. METHOD: Balb/C mice were subcutaneously injected with purified tubulin in different dosage of 100, 200 and 300 g and an equal valume of complete Freund’s adjuvant(CFA). Control mice underwent subcutaneous injection of PBS and CFA. Immunizations were boosted in incomplete Freund’s adjuvant (IFA) with 100, 200 and 300 g purified tubulin per groups twice. Control mice were given PBS in IFA at the contemporaneous boostertime as well. The ABR and DPOAE were recorded three times. The click and tone pips of 8, 16 and 32 kHz were generated to the ears twice after last boosting immunization. RESULT: Analysis of variance and t-test show significant difference in four weeks from 2 to 6 weeks of last boosting and between controls and tubulin-immunized groups after 2 and 6 weeks from last boosting. DPOAE of 100, 200 and 300 g tubulin immunization show abnormal result after 6 weeks from last booster immunization. Morphological study of temporal bone shows that in both 200 and 300 g tubulin immunized group we observe spiral ganglion damages as well as hair cells. CONCLUSION: This study showed that -tubulin is an autoantigen in animal model of hearing loss as well as in human autoimmune hearing loss patients.
J ALLERGY CLIN IMMUNOL FEBRUARY 2005
SATURDAY
Prevalence of Cystic Fibrosis in Children Who Present With Nasal Polyposis E. J. Schmitt, W. Neaville, T. Pongdee; Allergy/Immunology, University of Texas Southwestern Medical Center, Dallas, TX. RATIONALE: Children with cystic fibrosis have a higher incidence of nasal polyposis than unaffected children. Additionally, children with nasal polyposis are commonly screened for cystic fibrosis, whether or not other co-morbid features of cystic fibrosis are evident. The actual prevalence of cystic fibrosis diagnosed solely on the basis of isolated nasal polyposis disease has not been reported. METHODS: A medical records search identified all patients with an ICD-9 diagnosis code for cystic fibrosis and nasal polyps at Children’s Medical Center from 1999-2004. The order of each diagnosis was determined by chart review. RESULTS: We report 893 children with cystic fibrosis and 216 children with nasal polyps were seen at our institution over this five year period. Forty-five children were confirmed to have both diagnoses. Only 2 of 45 children developed nasal polyps as the initial manifestation that lead to the subsequent diagnosis of cystic fibrosis. Both of these subjects were otherwise healthy 6 year old Caucasian females, however one had chronic sinusitis with hypoplastic sinuses while the other was believed to have nasal allergies. We find a prevalence of 45/893 or 5% of cystic fibrosis patients with nasal polyps, but only 2/893 or 0.2% of the cystic fibrosis patients had their disease discovered because of nasal polyps. CONCLUSION: It is quite rare to diagnose cystic fibrosis secondary to the presence of nasal polyps. Furthermore, a suspicion for other etiologies aside from cystic fibrosis should be maintained in children who present with isolated nasal polyposis.
61
62
B Cell Superantigen-Induced Immune Complex Peritonitis
Anderson1,2,
Sporici1,
LaRosa2,
Levinson1,2; 1Philadel-
A. R. D. F. A. I. phia Veterans Administration Medical Center, Philadelphia, PA, 2University of Pennsylvania School of Medicine, Philadelphia, PA. RATIONALE: Staphylococcal protein A (SpA), a representative B cell superantigen (SAg), binds to the Fab regions of most VH3+ immunoglobulin molecules outside of their complementarity determining regions. We previously reported that the interaction of SpA with VH3+ immunoglobulin molecules produces a histologic pattern of inflammation in the skin of a rabbit indicative of immune complex injury. To begin to identify the cellular and molecular events contributing to this type of unconventional immune complex mediated inflammation, we established a mouse peritoneal Arthus model. METHODS: BALB/c mice were administered SpA intraperitoneally and either human polyclonal IgG (hIgG) or saline intravenously. Peritoneal lavage was performed 8 hours later and the fluid analyzed for differential white blood cell counts and levels of chemokines. VH3- depleted hIgG was substituted for hIgG in some experiments. Studies were also conducted in mast cell deficient and in TNF- knockout mice. RESULTS: BALB/c mice treated with SpA and hIgG developed a neutrophilic response that peaked eight hours after challenge. The response was dependent on the presence of VH3+ immunolgobulins. We also observed that mast cells and TNF- play obligatory roles in this reaction and that it is associated with the local release of the CXC chemokines MIP-2 and KC. CONCLUSIONS: These results provide further compelling evidence for the induction of immune-complex-mediated injury by a B cell SAg and
highlight important factors contributing to the pathogenesis of this novel type of inflammatory reaction. Funding: Veterans Administration Anti-Cyclic Citrullinated Peptide (CCP) Testing: Relevance in Rheumatoid Arthritis and Other Autoimmune Disorders J. M. Popov, M. Glovsky, K. Tokoro, O. Zhukov, F. Zarabpouri, K. Qu, A. Sferruzza; Immunology, Quest Diagnostics, San Juan Capistrano, CA. RATIONALE: CCP antibody is a relatively new, specific marker for detection of rheumatoid arthritis (RA) that can be used in conjunction with rheumatoid factor (RF) antibody. It is rarely present in other autoimmune diseases including systemic lupus erythematosus (SLE). In this study, we determined the frequency of CCP antibody in patients with RA and SLE as well as in patients with unknown or suspected autoimmune disease. METHODS: 516 sera sent to Quest Diagnostics for autoimmune testing (including ANA testing by EIA and IFA) and 113 sera from patients with RA (n=50) or SLE (n=63) were included. Presence of CCP antibody (IgG) was determined by enzyme-linked immunoassay. Further, presence of RF IgM was determined by latex agglutination. RESULTS: CCP antibody was present in 96% of RA cases and 3% of SLE cases. Thirty percent of the samples submitted for autoimmune testing (diagnosis unknown) were RF+/CCP+ and 70% were RF+/CCP-. Four of 120 (3%) ANA-positive samples were RF+/CCP+ and 27 (23%) were RF+/CCP-. CONCLUSIONS: The CCP antibody sensitivity in patients with RA is 96%. As expected, CCP antibody was rarely (3%) detected in patients with SLE. Patients with RF+/CCP- serotype are likely to have other diseases such as infection or immune complex diseases.
63
64
Immune Response in Tubulin Induces Hearing Loss in Mice
M. H. Kermany, Z. Bin, K. Mohammad, Z. Bin, J. Lee; Department of Medicine, University of Tennessee, Memphis, TN. RATIONALE: -Tubulin is an important molecule in the hair cells and supporting cells within the sensory epithelium and found to be an autoantigen in Méniere’s disease. Our goal is to induce hearing loss in mice with varying doses of -Tubulin antigen and evaluate the pathogenesis of autoimmune hearing loss induced by tubulin in mice. METHOD: Balb/C mice were subcutaneously injected with purified tubulin in different dosage of 100, 200 and 300 g and an equal valume of complete Freund’s adjuvant(CFA). Control mice underwent subcutaneous injection of PBS and CFA. Immunizations were boosted in incomplete Freund’s adjuvant (IFA) with 100, 200 and 300 g purified tubulin per groups twice. Control mice were given PBS in IFA at the contemporaneous boostertime as well. The ABR and DPOAE were recorded three times. The click and tone pips of 8, 16 and 32 kHz were generated to the ears twice after last boosting immunization. RESULT: Analysis of variance and t-test show significant difference in four weeks from 2 to 6 weeks of last boosting and between controls and tubulin-immunized groups after 2 and 6 weeks from last boosting. DPOAE of 100, 200 and 300 g tubulin immunization show abnormal result after 6 weeks from last booster immunization. Morphological study of temporal bone shows that in both 200 and 300 g tubulin immunized group we observe spiral ganglion damages as well as hair cells. CONCLUSION: This study showed that -tubulin is an autoantigen in animal model of hearing loss as well as in human autoimmune hearing loss patients.