Showing papers by "Erwin W. Gelfand published in 2014"

Newborn Screening for Severe Combined Immunodeficiency in 11 Screening Programs in the United States

Abstract: The purpose of newborn screening is early detection of inborn conditions for which prompt treatments mitigate mortality or irreversible damage. The first heritable immune disorders to which newborn screening has been applied are those that together comprise severe combined immunodeficiency (SCID), caused by defects in any of a diverse group of gene products essential for development of adaptive immunity provided by T and B lymphocytes.1,2 A feature of all SCID is defective production of T cells. In most SCID, B cells are also defective, but even normal B cells cannot produce antibodies without T-cell help. Thus, infants with SCID are susceptible to life-threatening infections. Early detection and treatment optimize survival.3-5 Provided that SCID is diagnosed before infections become overwhelming, affected infants can be rescued with hematopoietic stem cell transplantation; gene therapy; or, for adenosine deaminase deficiency, enzyme replacement therapy.2,5-8 Population-based screening is the only means to detect SCID prior to the onset of infections in most cases, as more than 80% lack a positive family history.9,10 T-cell receptor excision circles (TRECs), a biomarker for T lymphopoiesis,11 can be measured by polymerase chain reaction (PCR) using DNA isolated from infant dried blood spots collected for newborn screening.9 Dried blood spots from apparently healthy newborns who were later diagnosed with SCID lacked TRECs.9 Confirmation of the utility of the TREC test,12 adaptation for pilot newborn screening programs in Wisconsin13 and Massachusetts,14 and an evidence-based review led to the recommendation by the US Department of Health and Human Services Secretary in 2010 that SCID be added to the Uniform Screening Panel for all newborns, with related T-cell deficiencies added to the list of secondary targets.15 Currently, 23 states, the District of Columbia, and the Navajo Nation screen approximately two-thirds of all infants born in the United States for SCID. Individual states have confirmed detection of SCID as well as additional disorders with low T-cell numbers, which also may benefit from further assessment of immune dysfunction and from protective treatments.13,16-18 Here we present the first combined analysis of more than 3 million infants screened for SCID in 10 states and the Navajo Nation, providing a population-based overview of SCID and non-SCID T-cell lymphopenia.

Recommendations for live viral and bacterial vaccines in immunodeficient patients and their close contacts

Abstract: The present uncertainty of which live viral or bacterial vaccines can be given to immunodeficient patients and the growing neglect of societal adherence to routine immunizations has prompted the Medical Advisory Committee of the Immune Deficiency Foundation to issue recommendations based on published literature and the collective experience of the committee members. These recommendations address the concern for immunodeficient patients acquiring infections from healthy subjects who have not been immunized or who are shedding live vaccine–derived viral or bacterial organisms. Such transmission of infectious agents can occur within the hospital, clinic, or home or at any public gathering. Collectively, we define this type of transmission as close-contact spread of infectious disease that is particularly relevant in patients with impaired immunity who might have an infection when exposed to subjects carrying vaccine-preventable infectious diseases or who have recently received a live vaccine. Immunodeficient patients who have received therapeutic hematopoietic stem transplantation are also at risk during the time when immune reconstitution is incomplete or while they are receiving immunosuppressive agents to prevent or treat graft-versus-host disease. This review recommends the general education of what is known about vaccine-preventable or vaccine-derived diseases being spread to immunodeficient patients at risk for close-contact spread of infection and describes the relative risks for a child with severe immunodeficiency. The review also recommends a balance between the need to protect vulnerable subjects and their social needs to integrate into society, attend school, and benefit from peer education.

Is There a Link Between Obesity and Asthma

Abstract: Increasing epidemiological data identify a link between obesity and asthma incidence and severity. Based on experimental data, it is possible that shared inflammatory mechanisms play a role in determining this linkage. Although controversial, the role of adipokines may be central to this association and the maintenance of the asthma phenotype. While leptin and adiponectin have a causal link to experimental asthma in mice, data in humans are less conclusive. Recent studies demonstrate that adipokines can regulate the survival and function of eosinophils and that these factors can affect eosinophil trafficking from the bone marrow to the airways. In addition, efferocytosis, the clearance of dead cells, by airway macrophages or blood monocytes appears impaired in obese asthmatics and is inversely correlated with glucocorticoid responsiveness. This review examines the potential mechanisms linking obesity to asthma.

Somatic reversion in dedicator of cytokinesis 8 immunodeficiency modulates disease phenotype.

Abstract: Background Autosomal recessive loss-of-function mutations in dedicator of cytokinesis 8 (DOCK8) cause a combined immunodeficiency characterized by atopy, recurrent infections, and cancer susceptibility. A genotype-phenotype explanation for the variable disease expression is lacking. Objective We investigated whether reversions contributed to the variable disease expression. Methods Patients followed at the National Institutes of Health's Clinical Center were studied. We performed detailed genetic analyses and intracellular flow cytometry to detect DOCK8 protein expression within lymphocyte subsets. Results We identified 17 of 34 DOCK8-deficient patients who had germline mutations with variable degrees of reversion caused by somatic repair. Somatic repair of the DOCK8 mutations resulted from second-site mutation, original-site mutation, gene conversion, and intragenic crossover. Higher degrees of reversion were associated with recombination-mediated repair. DOCK8 expression was restored primarily within antigen-experienced T cells or natural killer cells but less so in naive T or B cells. Several patients exhibited multiple different repair events. Patients who had reversions were older and had less severe allergic disease, although infection susceptibility persisted. No patients were cured without hematopoietic cell transplantation. Conclusions In patients with DOCK8 deficiency, only certain combinations of germline mutations supported secondary somatic repair. Those patients had an ameliorated disease course with longer survival but still had fatal complications or required hematopoietic cell transplantation. These observations support the concept that some DOCK8-immunodeficient patients have mutable mosaic genomes that can modulate disease phenotype over time.

Janus kinase 1/3 signaling pathways are key initiators of TH2 differentiation and lung allergic responses

Abstract: Background Janus kinases (JAKs) are regulators of signaling through cytokine receptors. The importance of JAK1/3 signaling on T H 2 differentiation and development of lung allergic responses has not been investigated. Objective We sought to examine a selective JAK1/3 inhibitor (R256) on differentiation of T H subsets in vitro and on development of ovalbumin (OVA)–induced airway hyperresponsiveness (AHR) and inflammation in an experimental model of asthma. Methods A selective JAK1/3 inhibitor was used to assay the importance of this pathway on induction of T H 1, T H 2, and T H 17 differentiation in vitro . In vivo , the effects of inhibiting JAK1/3 signaling were examined by administering the inhibitor during the sensitization or allergen challenge phases in the primary challenge model or just before provocative challenge in the secondary challenge model. Airway inflammation and AHR were examined after the last airway challenge. Results In vitro , R256 inhibited differentiation of T H 2 but not T H 1 or T H 17 cells, which was associated with downregulation of signal transducer and activator of transcription (STAT) 6 and STAT5 phosphorylation. However, once polarized, T H 2 cells were unaffected by the inhibitor. In vivo , R256 administered during the OVA sensitization phase prevented the development of AHR, airway eosinophilia, mucus hypersecretion, and T H 2 cytokine production without changes in T H 1 and T H 17 cytokine levels, indicating that selective blockade of T H 2 differentiation was critical. Inhibitor administration after OVA sensitization but during the challenge phases in the primary or secondary challenge models similarly suppressed AHR, airway eosinophilia, and mucus hypersecretion without any reduction in T H 2 cytokine production, suggesting the inhibitory effects were downstream of T H 2 cytokine receptor signaling pathways. Conclusions Targeting the T H 2-dependent JAK/STAT activation pathway represents a novel therapeutic approach for the treatment of asthma.

Vaccine strain varicella-zoster virus–induced central nervous system vasculopathy as the presenting feature of DOCK8 deficiency

Abstract: To the Editor: In contrast to autosomal dominant forms of hyper-IgE syndrome resulting from mutations in the STAT3 gene, autosomal recessive dedicator of cytokinesis 8 (DOCK8) deficiency, results in susceptibility to cutaneous viral infections, eosinophilia, and allergic disease. CNS manifestations have been reported in patients with DOCK8 deficiency, but progressive multifocal leukoencephalopathy caused by JC virus has been considered the only known viral etiology to date (1, 2). A 6 year-old boy with atopy and recurrent peripheral blood eosinophilia developed acute intermittent vomiting, diarrhea, headache, and dizziness. Magnetic resonance imaging (MRI) of the brain was normal. One day later, he began giggling inappropriately, experienced left leg paresthesias, urinary incontinence, and fell upon attempting to stand. Repeat MRI with axial diffusion-weighted images revealed multiple areas of acute infarction in areas supplied by both anterior cerebral arteries, a branch of the anterior cerebral artery, the right posterior cerebral artery, and the left middle cerebral artery (Figure 1A). A complete blood count revealed a total eosinophil count of 2,160/μL, but no other abnormalities. He was treated with acetylsalicylic acid and intravenous methylprednisolone followed by oral prednisolone, 2 mg/kg for 3 days after which prednisolone was maintained at 1 mg/kg daily. Figure 1 Manifestation of vaccine strain varicella zoster virus-induced central nervous system vasculopathy On presentation to us one month later, he had developed paresthesias of his hands. Neurological examination showed only mild weakness in the hands. Computed tomography angiogram revealed diffuse vasculopathy. Subsequent 3-dimensional time of flight magnetic resonance angiographic imaging of the circle of Willis revealed vascular narrowing and post-stenotic dilatation (Figure 1B). Post-contrast T1W black blood arterial wall imaging illustrated avid enhancement and thickening of the distal supraclinoid internal carotid arterial walls bilaterally (Figure 1C). Cerebral spinal fluid (CSF) was obtained. While awaiting results of testing for viral CNS infections, the patient was treated with acyclovir, 30 mg/kg intravenous daily, and maintained on oral prednisolone, 2 mg/kg/day. The CSF at the time of diagnosis of vasculopathy contained 21 mononuclear cells and no red blood cells; protein was 39 mg/dL and glucose was 72 mg/dL. Quantitative PCR (Focus Diagnostics Reference Laboratory, Cypress, CA) amplified 7,116 copies of VZV DNA/mL in the CSF. To determine the VZV genotype, Forster Resonance Energy Transfer PCR was used to identify specific VZV DNA sequence polymorphisms within VZV open reading frames 38, 54, and 62, which distinguishes vaccine VZV from wild-type virus (Online Supplement Table 1) (3). This confirmed Oka varicella vaccine strain in the CSF. The CSF also contained anti-VZV IgG antibodies, but no antibodies to HSV-1 or HSV-2, enterovirus, cytomegalovirus, or Epstein-Barr virus were detected and the respective PCR analysis were similarly negative. The serum to CSF ratio of anti-VZV IgG antibody was markedly decreased (ratio 2.3) compared to albumin (ratio 149). Two weeks after treatment with acyclovir, the neurologic examination was completely normal, the CSF was acellular, and PCR was negative for VZV DNA. The patient received routine immunizations including VZV vaccination at 12 months of age. He had 3 episodes of vesicular rashes at ages 3, 4, and 5 years, which always occurred after completing a course of oral corticosteroids. The first rash tested positive for HSV. The second rash was identical. The third rash at age 5 years with vesicles on the lower thigh was diagnosed clinically as zoster. All rashes resolved on oral acyclovir. Past history included early-onset atopic dermatitis, food allergies often with anaphylaxis, biopsy-confirmed eosinophilic esophagitis, asthma and recurrent upper respiratory tract infections. Peripheral blood eosinophilia peaked at 9,000 eosinophils/μL and serum IgE at 472 IU/mL. During flares of respiratory or skin disease, he was treated with oral corticosteroids intermittently for 3 to 5 days, a few times annually. The frequency of such treatment increased and between ages 5½ and 6, he was treated with oral corticosteroids 1 to 2 times monthly during the prior 6 months. Conventional comparative genomic hybridization array analysis revealed a large deletion of exons 1 to 13 in a single allele of the DOCK8 gene. PCR analysis of genomic DNA and DOCK8 gene sequencing identified a single base pair mutation on the opposite allele at exon 12, resulting in a frame shift and premature stop codon: c.1266delC, p.Y423TfsX18, based upon reference sequence NM_00193536.1, isoform 3 (Figure 2A). Western Blot analysis confirmed lack of DOCK8 protein expression (Figure 2B). Parental testing demonstrated that the large exon deletion was inherited from the mother while the point mutation was inherited from the father. Immunological findings are summarized in Supplemental Table 2. Figure 2 DOCK8 molecular analyses The neurological symptoms and signs, imaging and CSF abnormalities, virological studies, and response to antiviral treatment were all features characteristically seen in VZV vasculopathy (4). Evidence of both focal and diffuse CNS disease was corroborated by widespread infarction produced by stenosis of multiple large cerebral arteries. The CSF examination revealed a pleocytosis as found in two-thirds of patients with VZV vasculopathy and both VZV DNA and anti-VZV IgG antibody with reduced serum/CSF ratios of anti-VZV IgG antibody were detected, indicative of intrathecal synthesis of anti-VZV IgG. Analysis of VZV DNA in CSF revealed that the VZV genotype was vaccine strain, demonstrating, for the first time, that VZV reactivation after vaccination in childhood can result in VZV vasculopathy. Although the underlying immunological consequences of DOCK8 deficiency remain to be fully elucidated, multiple immune system abnormalities may account for enhanced susceptibility to viral infection including impaired dendritic cell migration affecting T cell priming, lymphopenia, defective CD8 T cell activation and expansion, decreased production of the anti-viral cytokines IFN-γ and TNF-α, impaired T cell survival, decreased NK cell cytotoxicity, and antibody abnormalities (5-9). Germinal center formation and survival of germinal center B cells are impaired in DOCK8 deficiency, leading to defective long-lived antibody production (8). Responses to protein or polysaccharide-conjugated vaccines are often variable while responses to previously encountered viruses such as HSV and VZV have been normal as demonstrated here for VZV antibodies in serum and CSF. As in some other young patients with DOCK8 deficiency, this patient tended toward the milder spectrum of the disease with a relatively limited history of cutaneous infections, absence of severe systemic infections other than chronic mild otitis, and the modestly elevated serum IgE level. Instead, eosinophilia and moderate-severe eczema, asthma, and food sensitivities predominated. To control these conditions, courses of oral corticosteroids were administered with increased frequency over time. Interestingly, each of the 3 episodes of cutaneous viral infections with HSV or VZV was preceded by a course of oral corticosteroids. It is possible corticosteroids alone enabled the activation of vaccine strain VZV infection. More likely, the use of oral corticosteroids to gain disease control reduced the patient’s “immunologic threshold” and together resulted in reactivation of vaccine strain VZV and subsequent vasculopathy. In summary, a young patient with significant atopic disease and widespread infarction produced by stenosis of multiple large cerebral arteries was shown to express novel mutations on both alleles of the DOCK8 gene. For the first time, VZV vasculopathy was shown to be due to the vaccine strain. This case highlights the importance of considering the possibility of DOCK8 deficiency in the context of severe allergic disease and the potential risks for CNS infection including VZV vaccine-related vasculopathy.

Effects of Anti-G and Anti-F Antibodies on Airway Function after Respiratory Syncytial Virus Infection

Abstract: Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract illnesses in infants worldwide. Both RSV-G and RSV-F glycoproteins play pathogenic roles during infection with RSV. The objective of this study was to compare the effects of anti–RSV-G and anti–RSV-F monoclonal antibodies (mAbs) on airway hyperresponsiveness (AHR) and inflammation after primary or secondary RSV infection in mice. In the primary infection model, mice were infected with RSV at 6 weeks of age. Anti–RSV-G or anti–RSV-F mAbs were administered 24 hours before infection or Day +2 postinfection. In a secondary infection model, mice were infected (primary) with RSV at 1 week (neonate) and reinfected (secondary) 5 weeks later. Anti–RSV-G and anti–RSV-F mAbs were administered 24 hours before the primary infection. Both mAbs had comparable effects in preventing airway responses after primary RSV infection. When given 2 days after infection, anti–RSV-G–treated mice showed significantly decreased AHR and airway inflammati...

Role of prostaglandin D2 and CRTH2 blockade in early- and late-phase nasal responses.

Abstract: SummaryBackground Prostaglandin D2 (PGD2) plays an important role in allergic inflammation. The PGD2 receptor, CRTH2, is expressed on basophils, eosinophils, and Th2 lymphocytes and mediates chemotactic activity. Objective To define the role of CRTH2 in allergen-induced nasal responses in a mouse model of allergic rhinitis (AR), a potent, selective CRTH2 receptor antagonist, ARRY-063 was administered in a model of allergic rhinitis in mice. Methods ARRY-063 was administered orally to ovalbumin (OVA) sensitized and challenged mice. To assess nasal obstruction, respiratory frequency (RF) was monitored by whole-body plethysmography immediately after the 4th challenge (early-phase response, EPR) and 24 h after the 6th challenge (late-phase response, LPR). Nasal resistance (RNA) was also measured in the LPR. PGD2 was administered with or without OVA to determine the effect of PGD2 on nasal responsiveness. Cytokine levels and histopathological changes in nasal tissue were analysed. Results Instillation of PGD2 in the nose of sensitized mice together with a low concentration of OVA induced both an EPR and LPR. Treatment with the CRTH2 receptor antagonist prevented the decreases in RF seen immediately following the 4th challenge of sensitized mice (EPR). In the LPR, decreases in RF and increases in RNA were also prevented by antagonist treatment associated with reduced cytokine levels and inflammation in nasal tissues. Conclusions These data identify PGD2 as a mediator of both the EPR and LPR in this model of AR and suggest that antagonism of CRTH2 prevents the development of both the EPR and LPR as well as nasal inflammation.

Leukotriene B4 receptor 1 is differentially expressed on peripheral T cells of steroid-sensitive and -resistant asthmatics

Abstract: Background Numbers of CD8 + T cells expressing the leukotriene B 4 (LTB4) receptor, BLT1, have been correlated with asthma severity. Objective To examine the activation and numbers of BLT1-expressing peripheral blood CD4 + and CD8 + T cells from patients with steroid-sensitive (SS) and steroid-resistant (SR) asthma. Methods CD4 + and CD8 + T cells isolated from peripheral blood of healthy human subjects and patients with SS and SR asthma were stimulated in culture with anti-CD3/anti-CD28 followed by analysis of BLT1 surface expression and cytokine production. Activation of CD8 + T cells after ligation of BLT1 by LTB4 was monitored by changes in intracellular Ca 2+ concentrations. Results The number of BLT1-expressing cells was larger in patients with asthma than in controls and larger on activated CD8 + than on CD4 + T cells. Addition of LTB4 to activated CD8 + T cells resulted in increases in intracellular Ca 2+ concentrations. Expansion of activated CD4 + T cells, unlike CD8 + T cells, was significantly decreased in the presence of corticosteroid. In patients with SS asthma, numbers of BLT1-expressing CD8 + T cells were lower in the presence of corticosteroid, unlike in those with SR asthma in whom cell expansion was maintained. Levels of interleukin-13 were highest in cultured CD8 + T cells, whereas interleukin-10 levels were higher in CD4 + T cells from controls and patients with SS asthma. Interferon-γ levels were lowest in patients with SR asthma. Conclusion Differences in BLT1 expression, steroid sensitivity, and cytokine production were demonstrated in T lymphocytes from patients with SS and SR asthma. The LTB4–BLT1 pathway in CD8 + cells may play an important role in asthma and serve as an important target in the treatment of patients with SR asthma.

Successful hematopoietic cell transplantation in patients with unique NF-κB essential modulator (NEMO) mutations.

Abstract: Successful hematopoietic cell transplantation in patients with unique NF-κB essential modulator (NEMO) mutations

JNK2 Regulates the Functional Plasticity of Naturally Occurring T Regulatory Cells and the Enhancement of Lung Allergic Responses

Abstract: Glucocorticoid-induced TNFR family–related protein (GITR)–mediated activation of JNK was shown to regulate the suppressive activity of CD4 + CD25 + naturally occurring T regulatory cells (nTregs) in wild-type (WT) hosts. In this study, CD4 + CD25 + T cells were shown to be capable of becoming pathogenic effector cells in sensitized and challenged CD8 −/− recipient mice. Only GITR-expressing CD4 + CD25 + T cells, but neither GITR knocked-in CD4 + CD25 − T cells nor GITR-silenced CD4 + CD25 + T cells, enhanced development of lung allergic responses. Inhibition of JNK in WT nTregs or nTregs from GITR −/− and JNK2 −/− mice failed to enhance lung allergic responses in sensitized and challenged CD8 −/− recipient mice. The failure to enhance responses was associated with increased bronchoalveolar lavage fluid levels of IL-10 and TGF-β and decreased levels of IL-5, IL-6, and IL-13. In contrast, nTregs from JNK1 −/− mice, similar to WT nTregs, were fully effective in enhancing responses. Thus, GITR stimulation of nTregs and signaling through JNK2, but not JNK1, triggered the loss of regulatory function while concomitantly gaining pathogenic CD4 + T effector cell function responsible for exacerbating asthma-like immunopathology.

Modulated expression of genes encoding estrogen metabolizing enzymes by G1-phase cyclin-dependent kinases 6 and 4 in human breast cancer cells.

Abstract: G1-phase cell cycle defects, such as alterations in cyclin D1 or cyclin-dependent kinase (cdk) levels, are seen in most tumors. For example, increased cyclin D1 and decreased cdk6 levels are seen in many human breast tumors. Overexpression of cdk6 in breast tumor cells in culture has been shown to suppress proliferation, unlike the growth stimulating effects of its close homolog, cdk4. In addition to directly affecting proliferation, alterations in cdk6 or cdk4 levels in breast tumor cells also differentially influence levels of numerous steroid metabolic enzymes (SMEs), including those involved in estrogen metabolism. Overexpression of cdk6 in tumor cell lines having low cdk6 resulted in decreased levels of mRNAs encoding aldo-keto reductase (AKR)1C1, AKR1C2 and AKR1C3, which are hydroxysteroid dehydrogenases (HSDs) involved in steroid hormone metabolism. In contrast, increasing cdk4 dramatically increased these transcript levels, especially those encoding AKR1C3, an enzyme that converts estrone to 17β-estradiol, a change that could result in a pro-estrogenic state favoring tumor growth. Effects on other estrogen metabolizing enzymes, including cytochrome P450 (CYP) 19 aromatase, 17β-HSD2, and CYP1B1 transcripts, were also observed. Interactions of cdk6 and cdk4, but not cyclin D1, with the promoter region of a cdk-regulated gene, 17β-HSD2, were detected. The results uncover a previously unsuspected link between the cell cycle and hormone metabolism and differential roles for cdk6 and cdk4 in a novel mechanism for pre-receptor control of steroid hormone action, with important implications for the origin and treatment of steroid hormone-dependent cancers.

Targeting Pim1 kinase in the treatment of peanut allergy.

Abstract: Introduction: Peanut sensitization is the most significant food allergen associated with life-threatening allergic reactions. Unlike many food sensitivities, peanut allergy often persists into adulthood. Presently, the only effective therapy is peanut avoidance. Effective preventative therapy requires an understanding of the pathways that lead to anaphylaxis. IgE and mast cell activation are essential contributors. The responsible pathways upstream are driven by pro-allergic T helper 2 differentiation and release of cytokines including interleukin-4 (IL-4) and IL-13. Areas covered: The research utilized an experimental model of peanut-induced anaphylaxis in mice that mimics many of the responses seen in the human disease. Following peanut sensitization and challenge, clinical responses, intestinal inflammatory and immune cell interactions, and genetic and molecular events were monitored. For the first time, evidence for Pim1 kinase involvement was demonstrated in association with the downregulation of Run...

Differential DNA Methylation In Mothers Increases The Prevalence Of Atopic Dermatitis In Their Offspring

Abstract: S U N D A Y 522 Differential DNA Methylation In Mothers Increases The Prevalence Of Atopic Dermatitis In Their Offspring Dr. Pia J. Hauk, MD, Mrs. Anna Forssen, MS, Dr. Brent Pedersen, PhD, Dr. Matthew Strand, PhD, Mrs. Lindsay Munoz, BA, Dr. Michaela Schedel, PhD, Dr. Anne Lynch, MD, MSPH, Dr. Virginia Winn, MD, PhD, David A. Schwartz, MD, Dr. Erwin W. Gelfand, MD, FAAAAI; National Jewish Health, Denver, CO, University of Colorado Denver, Aurora, CO, University of Colorado Denver School of Medicine, Aurora, CO. RATIONALE: Allergic diseases are increasing in developed countries with early manifestations as atopic dermatitis and food sensitivities. To determine the effects of prenatal diet on DNA methylation and transgenerational inheritance of atopy, we studied a birth cohort of children born to mothers at high and at a low risk of having an atopic child based on parental history. METHODS: Three hundred three newborns born to 159 highand 144 low-risk mothers were followed biannually for two years for development of atopy. Genome-wide DNA methylation patterns were determined on maternal peripheral blood mononuclear cells (PBMC) using Comprehensive High-throughput Arrays for Relative Methylation (CHARM). Prenatal intake (NIH NCI Diet History Questionnaire) and serum concentrations of the methyl donors folate and vitamin B12 were measured. RESULTS: Atopic dermatitis wasmore prevalent in children ofmothers at high risk of atopy compared to children of low-risk mothers at age 6 months (36.1% versus 20.9%; p50.0096), 12 months (41.3% versus 17.9%; p50.0001), 18 months (31.4% versus 17.4%; p50.023) and 24 months (38.8% versus 15.6%; p50.0119). Greater than 95% of mothers in both groups took prenatal preparations containing folate. While dietary intake and serum levels of folate and vitamin B12 did not differ between highand low-risk mothers, DNAmethylation analyses showed significant differences in PBMC from highand low-risk mothers, with 64 gene loci being differentially methylated (increasedmethylation at 22 and decreased methylation at 42 loci). CONCLUSIONS: These data suggest that DNA methylated/demethylated gene loci in mothers at high-risk may affect the development of atopy in their offspring.

Targeting the Steroidogenic Pathway For Treating and/or Preventing Allergic Diseases

Abstract: The present invention relates to methods and compositions for treating and/or preventing allergic diseases or conditions by inhibiting one or more components of the steroidogenic pathway.