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Interferon-Gamma Receptor 1 Gene Polymorphisms in Patients with Herpes Simplex Virus Infection or Lyme Disease
S14 Abstracts
SATURDAY
37
Omenn Syndrome is Associated with Mutations in DNA Ligase IV V. H. D. Kim, E. Grunebaum, A. Bates, C. M. Roifman; Hospital for Sick Children, Toronto, ON, Canada. RATIONALE: Omenn syndrome (OS) is a variant of severe combined immunodeficiency (SCID) that is fatal unless treated with allogeneic bone marrow transplant (BMT). We report the first case of a patient with mutations in DNA Ligase IV gene who presented with typical clinical and laboratory features of OS, which was corrected after allogeneic BMT. METHODS: A review of a case of OS and analysis of DNA Ligase IV gene. RESULTS: A 3 week old female born to non-consanguineous parents presented with protracted erythroderma, hepatosplenomegaly, generalized lymphadenopathy and diarrhea. Evaluation of the immune system revealed markedly elevated eosinophils, low CD3 1 T cells, markedly reduced T cell response to mitogen stimulation and markedly reduced T cell receptor excision circles (TREC). Analysis of T cell receptor (TCR) V beta families showed excessive expansion of 3 TCR families and underrepresentation of most other families. B cells were nearly absent and serum IgA and IgM levels were reduced. Sequencing of DNA Ligase IV gene revealed 3 heterozygous mutations, including a novel deletion. At 6 months of age, the patient received an HLA-matched unrelated donor BMT following myeloablative conditioning with busulphan and cyclophosphamide. At 3.5 years after transplant, the patient continues to demonstrate complete donor chimerism, and T and B cell numbers, T cell responses to mitogens, TREC, immunoglobulin production and TCR repertoire were all normal. CONCLUSIONS: Patients with mutations in DNA Ligase IV can present with features of OS. BMT with full myeloablative conditioning can be effectively and safely used to correct the SCID.
38
Quantitation of Antibodies in the Sera of Immunodeficient Patients Receiving Monthly Intravenous Immune Globulin (IVIg) M. M. Pieretti, C. Cunningham-Rundles; Mount Sinai, New York, NY. RATIONALE: Immunodeficient patients receiving IVIg are presumed to possess sufficient passive immunity to a variety of microbial antigens. The goal of this study was to determine if the level of antibodies would be considered in the protective range. METHODS: Sera were obtained from 7 patients with X-linked Agammaglobulinemia (XLA), 1 with Hyper-IgM syndrome (HIGM) and 13 with Common Variable Immune Deficiency (CVID) just before infusions. To minimize contributions from endogenous IgG, subjects with very low levels were chosen: the median initial IgG was 50 mg/dL, with a range 10-202 mg/dL. Levels of IgG to Streptococcus pneumoniae, Haemophilus influenzae type b (Hib), tetanus, diphtheria, varicella-zoster (VZV), measles, mumps and rubella were compared to laboratory values considered protective. Five products of IVIg were in use; IgG trough values were >600 mg/dL. RESULTS: All sera had laboratory defined-protective antibodies to Hib (IgG 0.15 mg/mL; range: 0.9-6.43), measles (IgG 0.7; 1.16-2.85), mumps (IgG 0.5; 0.82-2.1), rubella (IgG 10 IU/mL; 71.2->500), VZV (IgG 0.9; 1.44-2.61) and diphtheria (IgG 0.1 IU/mL; 0.241.25). However, only 13 of 21 patients (62%) had adequate titers (IgG > 0.49 IU/mL) to tetanus. For pneumococcus, 11 of 20 patients (55%) had levels considered protective (IgG 1.3 mg/mL) for 7 of 14 serotypes tested. None of the sera had laboratory defined protective antibodies to certain pneumococcal serotypes, such as 4, 9N and 12F. CONCLUSIONS: Patients receiving monthly IVIg demonstrate adequate antibody titers to Hib, measles, mumps, rubella, VZV and diphtheria. However, a significant number lack protective levels of antibody to tetanus and pneumococci despite adequate trough IgG levels.
J ALLERGY CLIN IMMUNOL FEBRUARY 2009
39
B-Lymphocyte Deficiency and Neutropenia in a Patient with Normal Immunoglobulins S. A. Stutes1, K. Beane2, P. Mustillo1; 1Nationwide Children’s Hospital, Ohio State Univerisity, Columbus, OH, 2Nationwide Children’s Hospital, Columbus, OH. RATIONALE: B-cell deficient patients typically present early in life with sinopulmonary, skin, or enteroviral infections related to hypogammaglobulinemia. Reactive neutropenia may also be a feature. The finding of hypogammaglobulinemia in males typically prompts immunophenotyping, which may reveal a B-cell deficiency. This most commonly results from X-linked mutations in Bruton’s tyrosine kinase(BTK). We report a 4 year old male with absent B-cells with clinical phenotype similar to X-linked agammaglobulinemia (XLA) despite normal quantitative immunoglobulin (Ig) levels. METHODS: Description of clinical presentation and laboratory studies including immunophenotyping, Ig’s, BM biopsy, LAD markers, mitogen studies, HIV, and NBT to rule out alternative diagnosis. RESULTS: 4 year-old Senegalese male with a history of non-purulent abscesses, aseptic pericarditis, and neutropenia presented with a protracted enteroviral meningoencephalitis illness. Studies revealed normal NBT, LAD markers, mitogen studies, and BM biopsy. Ig’s were normal (IgG573, IgA-372, IgM-22, IgE-12) with normal dip/tet antibody titers, but low HIB and pneumococcal titers following primary vaccination series. Immunophenotyping demonstrated undetectable CD10/19/20/40 B-cell markers. BTK genetic analysis was normal. High dose IVIg led to resolution of acute enteroviral CNS infection, with no further infections for the subsequent 18 months on monthly IVIg. Recently his IgM levels have dropped. CONCLUSIONS: This case demonstrates the need to consider diagnosis of Bruton’s agammaglobulinemia despite normal immunoglobulin levels in males with a suspicious history, especially when associated with neutropenia. This represents an atypical Bruton’s patient, initially with normal immunoglobulins, which may be due to a small number of B-cells present mostly in lymphoid tissue but absent in peripheral circulation.
40
Interferon-Gamma Receptor 1 Gene Polymorphisms in Patients with Herpes Simplex Virus Infection or Lyme Disease L. P. Titov1, O. O. Yanovich1, V. V. Scherba1, L. M. DuBuske2; 1Research Institute for Epidemiology and Microbiology, Minsk, Belarus, 2Immunology Research Institute of New England, Gardner, MA. RATIONALE: Interferon-gamma is a key mediator of immune responses. The receptor for IFN-gamma is made up of alpha and beta-subunits, both of which are integral membrane proteins. The alpha subunit, encoded by the gene IFNGR1, plays a critical role in ligand binding and signal transduction. This study assesses SNP polymorphisms of the IFNGR1 gene in patients with Herpes Simplex Virus (HSV) and Lyme Disease. METHODS: The study included a control group (n 5 40) of healthy subjects, and patients with HSV (n 5 23) or Lyme disease (n 5 26). The role of two polymorphisms in the IFNGR1 gene (SNPs 195, -270) were assessed by allele-specific PCR. RESULTS: There were significant differences between the control group and the HSV patients in frequency of IFNGR1 1 95 genotypes. The frequency of genotype TT was increased in HSV patients (52%) compared with the control group (12.5%; p < 0.05). There was a significant association between genotype CC for SNP 195 in patients with Lyme Disease compared with the control group (42% versus 20%; p< 0.05). There was no statistically significant difference in SNP -270 IFNGR1 genotype frequencies between the studied groups. CONCLUSIONS: Patients with HSV and Lyme Disease have a genetic defect in the IFNGR1 gene in SNP 195 which lies at the beginning of intron 1 and in close proximity to a splicing site. Changes in the nucleotide at the first intron can increase or decrease the binding of a particular transcription factor and ultimately the export of cytokine product, suggesting that this SNP may have a role in disease susceptibility or progression.
SATURDAY
37
Omenn Syndrome is Associated with Mutations in DNA Ligase IV V. H. D. Kim, E. Grunebaum, A. Bates, C. M. Roifman; Hospital for Sick Children, Toronto, ON, Canada. RATIONALE: Omenn syndrome (OS) is a variant of severe combined immunodeficiency (SCID) that is fatal unless treated with allogeneic bone marrow transplant (BMT). We report the first case of a patient with mutations in DNA Ligase IV gene who presented with typical clinical and laboratory features of OS, which was corrected after allogeneic BMT. METHODS: A review of a case of OS and analysis of DNA Ligase IV gene. RESULTS: A 3 week old female born to non-consanguineous parents presented with protracted erythroderma, hepatosplenomegaly, generalized lymphadenopathy and diarrhea. Evaluation of the immune system revealed markedly elevated eosinophils, low CD3 1 T cells, markedly reduced T cell response to mitogen stimulation and markedly reduced T cell receptor excision circles (TREC). Analysis of T cell receptor (TCR) V beta families showed excessive expansion of 3 TCR families and underrepresentation of most other families. B cells were nearly absent and serum IgA and IgM levels were reduced. Sequencing of DNA Ligase IV gene revealed 3 heterozygous mutations, including a novel deletion. At 6 months of age, the patient received an HLA-matched unrelated donor BMT following myeloablative conditioning with busulphan and cyclophosphamide. At 3.5 years after transplant, the patient continues to demonstrate complete donor chimerism, and T and B cell numbers, T cell responses to mitogens, TREC, immunoglobulin production and TCR repertoire were all normal. CONCLUSIONS: Patients with mutations in DNA Ligase IV can present with features of OS. BMT with full myeloablative conditioning can be effectively and safely used to correct the SCID.
38
Quantitation of Antibodies in the Sera of Immunodeficient Patients Receiving Monthly Intravenous Immune Globulin (IVIg) M. M. Pieretti, C. Cunningham-Rundles; Mount Sinai, New York, NY. RATIONALE: Immunodeficient patients receiving IVIg are presumed to possess sufficient passive immunity to a variety of microbial antigens. The goal of this study was to determine if the level of antibodies would be considered in the protective range. METHODS: Sera were obtained from 7 patients with X-linked Agammaglobulinemia (XLA), 1 with Hyper-IgM syndrome (HIGM) and 13 with Common Variable Immune Deficiency (CVID) just before infusions. To minimize contributions from endogenous IgG, subjects with very low levels were chosen: the median initial IgG was 50 mg/dL, with a range 10-202 mg/dL. Levels of IgG to Streptococcus pneumoniae, Haemophilus influenzae type b (Hib), tetanus, diphtheria, varicella-zoster (VZV), measles, mumps and rubella were compared to laboratory values considered protective. Five products of IVIg were in use; IgG trough values were >600 mg/dL. RESULTS: All sera had laboratory defined-protective antibodies to Hib (IgG 0.15 mg/mL; range: 0.9-6.43), measles (IgG 0.7; 1.16-2.85), mumps (IgG 0.5; 0.82-2.1), rubella (IgG 10 IU/mL; 71.2->500), VZV (IgG 0.9; 1.44-2.61) and diphtheria (IgG 0.1 IU/mL; 0.241.25). However, only 13 of 21 patients (62%) had adequate titers (IgG > 0.49 IU/mL) to tetanus. For pneumococcus, 11 of 20 patients (55%) had levels considered protective (IgG 1.3 mg/mL) for 7 of 14 serotypes tested. None of the sera had laboratory defined protective antibodies to certain pneumococcal serotypes, such as 4, 9N and 12F. CONCLUSIONS: Patients receiving monthly IVIg demonstrate adequate antibody titers to Hib, measles, mumps, rubella, VZV and diphtheria. However, a significant number lack protective levels of antibody to tetanus and pneumococci despite adequate trough IgG levels.
J ALLERGY CLIN IMMUNOL FEBRUARY 2009
39
B-Lymphocyte Deficiency and Neutropenia in a Patient with Normal Immunoglobulins S. A. Stutes1, K. Beane2, P. Mustillo1; 1Nationwide Children’s Hospital, Ohio State Univerisity, Columbus, OH, 2Nationwide Children’s Hospital, Columbus, OH. RATIONALE: B-cell deficient patients typically present early in life with sinopulmonary, skin, or enteroviral infections related to hypogammaglobulinemia. Reactive neutropenia may also be a feature. The finding of hypogammaglobulinemia in males typically prompts immunophenotyping, which may reveal a B-cell deficiency. This most commonly results from X-linked mutations in Bruton’s tyrosine kinase(BTK). We report a 4 year old male with absent B-cells with clinical phenotype similar to X-linked agammaglobulinemia (XLA) despite normal quantitative immunoglobulin (Ig) levels. METHODS: Description of clinical presentation and laboratory studies including immunophenotyping, Ig’s, BM biopsy, LAD markers, mitogen studies, HIV, and NBT to rule out alternative diagnosis. RESULTS: 4 year-old Senegalese male with a history of non-purulent abscesses, aseptic pericarditis, and neutropenia presented with a protracted enteroviral meningoencephalitis illness. Studies revealed normal NBT, LAD markers, mitogen studies, and BM biopsy. Ig’s were normal (IgG573, IgA-372, IgM-22, IgE-12) with normal dip/tet antibody titers, but low HIB and pneumococcal titers following primary vaccination series. Immunophenotyping demonstrated undetectable CD10/19/20/40 B-cell markers. BTK genetic analysis was normal. High dose IVIg led to resolution of acute enteroviral CNS infection, with no further infections for the subsequent 18 months on monthly IVIg. Recently his IgM levels have dropped. CONCLUSIONS: This case demonstrates the need to consider diagnosis of Bruton’s agammaglobulinemia despite normal immunoglobulin levels in males with a suspicious history, especially when associated with neutropenia. This represents an atypical Bruton’s patient, initially with normal immunoglobulins, which may be due to a small number of B-cells present mostly in lymphoid tissue but absent in peripheral circulation.
40
Interferon-Gamma Receptor 1 Gene Polymorphisms in Patients with Herpes Simplex Virus Infection or Lyme Disease L. P. Titov1, O. O. Yanovich1, V. V. Scherba1, L. M. DuBuske2; 1Research Institute for Epidemiology and Microbiology, Minsk, Belarus, 2Immunology Research Institute of New England, Gardner, MA. RATIONALE: Interferon-gamma is a key mediator of immune responses. The receptor for IFN-gamma is made up of alpha and beta-subunits, both of which are integral membrane proteins. The alpha subunit, encoded by the gene IFNGR1, plays a critical role in ligand binding and signal transduction. This study assesses SNP polymorphisms of the IFNGR1 gene in patients with Herpes Simplex Virus (HSV) and Lyme Disease. METHODS: The study included a control group (n 5 40) of healthy subjects, and patients with HSV (n 5 23) or Lyme disease (n 5 26). The role of two polymorphisms in the IFNGR1 gene (SNPs 195, -270) were assessed by allele-specific PCR. RESULTS: There were significant differences between the control group and the HSV patients in frequency of IFNGR1 1 95 genotypes. The frequency of genotype TT was increased in HSV patients (52%) compared with the control group (12.5%; p < 0.05). There was a significant association between genotype CC for SNP 195 in patients with Lyme Disease compared with the control group (42% versus 20%; p< 0.05). There was no statistically significant difference in SNP -270 IFNGR1 genotype frequencies between the studied groups. CONCLUSIONS: Patients with HSV and Lyme Disease have a genetic defect in the IFNGR1 gene in SNP 195 which lies at the beginning of intron 1 and in close proximity to a splicing site. Changes in the nucleotide at the first intron can increase or decrease the binding of a particular transcription factor and ultimately the export of cytokine product, suggesting that this SNP may have a role in disease susceptibility or progression.