Nesrine A. Mohamed

Influence of glucocorticoid receptor gene NR3C1 646 C>G polymorphism on glucocorticoid resistance in asthmatics: a preliminary study

Background Glucocorticoid receptor gene polymorphism (NR3C1 646 C>G) may play an important role in the development of severe bronchial asthma and resistance to glucocorticoids (GCs). Objective The aim of the present study was to determine the relation between the 646 C>G polymorphism of the glucocorticoid receptor gene (NR3C1) and resistance to GCs with development of severe bronchial asthma. Material and methods This case-control study included 40 patients with severe bronchial asthma and 20 apparently healthy controls. Atopic status was determined by skin prick test reaction to the most common locally-encountered allergens. GCs reversibility test was performed to differentiate between GCs sensitive and GCs resistant asthma. For all subjects, analysis of the glucocorticoid receptor gene polymorphism (NR3C1 646 C>G) was done using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Results The frequencies of NR3C1 646 C>G genotypes and alleles differed significantly between asthmatic patients and controls. The frequencies of the CC genotype and C allele carriers were significantly higher among asthmatics than among controls, and also among GCs sensitive asthmatics than among GCs resistant asthmatics. However, NR3C1 646 C>G genotypes and alleles frequencies did not differ significantly according to the atopic status in asthmatics. Conclusions The too small sized of the investigated groups is a shortcoming of this study. Nevertheless, the observed variations demonstrate a marked association of NR3C1 646 C>G CC genotype with the development of bronchial asthma and a higher frequency of the C allele among GCs sensitive asthmatics. Large-scale studies are required to investigate the association between polymorphisms of the NR3C1 gene and GCs resistance among asthmatic patients.

OX40 ligand: a potential costimulatory molecule in atopic asthma.

Abstract Objective: We aimed to assess the percentage of peripheral blood B-lymphocytes expressing OX40 ligand (OX40L) in adult atopic and non-atopic asthmatic patients, and in healthy controls. Methods: This case–control study included 15 atopic asthmatic patients, 15 non-atopic asthmatic patients, and 15 healthy controls. Atopic status was determined by skin prick test reaction to the most common locally-encountered allergens. For all subjects, pulmonary function tests and measurement of total serum immunoglobulin E (IgE) levels by ELISA were performed. In addition, the percentage of B-lymphocytes expressing OX40L was assessed by flow cytometry in all three groups. Results: OX40L expression was significantly higher in atopic asthmatics than in non-atopic asthmatics and controls, but did not differ significantly between non-atopic asthmatics or controls. Among atopic asthmatics, OX40L expression correlated positively with total serum IgE levels, but not with age, disease duration, or values of forced expiratory volume in the first second. Conclusion: The over-expression of OX40L in atopic asthmatic patients appears to be linked to markers of the atopic status as total serum IgE, and signifies the vital role of OX40L in the atopic mechanism. Further large-scale studies are needed to investigate the role of OX40L in other atopic diseases and its relation to disease activity and severity.

Comparison between magnetic activated cell sorted monocytes and monocyte adherence techniques for in vitro generation of immature dendritic cells: an Egyptian trial.

Introduction Dendritic cells (DCs) are the most efficient antigen presenting cells, which are considered a central component of the immune system for their extraordinary capacity to initiate and modulate the immune responses elicited upon recognition of infectious agents. This has made them a major focus of interest in the conception of immunotherapeutic vaccine strategies. Aim of the study To standardise a protocol for in vitro differentiation of human peripheral blood monocytes into immature DCs (iDCs) upon treatment with specific growth factors and to compare two monocyte isolation methods including magnetic activated cell sorted (MACS) monocytes by CD14+ immuno-magnetic beads and monocytes separated by adherence. Material and methods Immature DCs were generated from monocytes of human peripheral blood in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 after in vitro culture for seven days. Cultured cells were stained with surface markers of iDCs: FITC-anti-CD14, PE-anti-CD11c, PE-anti-CD1a, PE-Cy5-anti-HLA-DR, and PE-anti-CD83 for flow cytometry analysis. Results We found that the viability of MACS-DCs was higher than DCs derived from monocytes separated by adherence (median 50 and interquartile range 45-50 vs. 25 and 10-30, respectively; p < 0.001). Flow cytometry analysis revealed that the median interquartile percentages of MACS-DCs expressing CD14– was significantly higher compared to the DCs derived from monocytes separated by adherence (median 80.2 and interquartile range 77.7-80.7 vs. 40.2 and 30.4-40.6, respectively; p < 0.001). However, MACS-DCs expressed the same levels of CD11c, CD1a, and HLA-DR as well as CD83 compared to the DCs derived from monocytes separated by adherence with p value > 0.05. Conclusions Both positively selected monocytes and monocytes separated by adherence procedure gave the same results as regards cell surface marker expression, although the DCs purity and viability using MACS separated monocytes were better.

The immunomodulatory role of zinc in asthmatic patients

HighlightsHypozincemia, among atopic asthmatics, was significantly linked to total serum IgE.Hypozincemia, among atopic asthmatics, was associated with alterations in the Th1 cytokine profile (IFN‐&ggr;), and the Th2 cytokine profile (IL‐10).Hypozincemia might play a role in the pathogenesis of atopic asthma.Zinc supplements might be useful in the treatment and prevention of atopic asthma. Background: Zinc deficiency may play an important role in the development of atopic asthma. The aim of the work: To assess serum zinc levels in adult atopic, non‐atopic asthmatic patients, and in healthy controls and to investigate its modulatory effect on production of interferon gamma (IFN‐&ggr;) and interleukin‐10 (IL‐10) by peripheral blood mononuclear cells (PBMCs) in vitro. Methods: Sixty asthmatics and 30 apparently healthy volunteers were included in this study. All patients were subjected to history taking, clinical examination, pulmonary function tests, skin prick test (SPT), serum zinc assessment by a colorimetric method as well as serum total IgE measurement by Enzyme‐linked immunosorbent assay (ELISA). PBMCs were activated in vitro in the presence and absence of zinc, and then cell culture supernatants were analyzed for IFN‐&ggr; and IL‐10 by ELISA. Results: Serum zinc levels were significantly lower in atopic asthmatics than non‐atopic asthmatics and healthy controls. In atopic asthmatics, highly significant correlations were found between zinc levels and total Ig E levels as well as FEV1. In culture, zinc triggers IFN‐&ggr; and inhibits IL‐10 production by PBMCs, in atopic asthmatics. In non atopic asthmatics and healthy controls, IFN‐&ggr; and IL‐10 were slightly affected by zinc supplementation in culture. Conclusion: Serum zinc levels affect asthma phenotypes. Atopic asthmatics might benefit from zinc supplements.

Transient elastography as a noninvasive assessment tool for hepatopathies of different etiology in pediatric type 1 diabetes mellitus

AIM To identify the prevalence and effect of hepatopathies of different etiologies among pediatric patients with type 1 diabetes mellitus (T1DM) using transient elastography (TE) and its relation to glycemic control. METHODS One hundred T1DM patients were studied focusing on liver functions, fasting lipid profile, hemoglobin A1c (HbA1c), hepatitis C virus (HCV), serum immunoglobulins, autoimmune antibodies; anti-nuclear antibody (ANA), anti-smooth muscle antibody (ASMA), and anti-liver kidney microsomal antibody (anti-LKM). Abdominal ultrasound was performed and TE was done for patients with HCV, positive autoimmune antibody and/or abnormal ultrasound findings. RESULTS Thirty-one patients were found to have one or more hepatic abnormalities; clinical hepatomegaly in 8%, elevated alanine aminotransferase (ALT) in 10%, HCV in 6%, autoimmune hepatitis (AIH) in 11% (10 were positive for ASMA and 2 were positive for ANA while anti-LKM antibodies were negative) and abnormal hepatic ultrasound in 20% (12 non-alcoholic fatty liver disease, 5 AIH, 2 HCV, 1 Mauriac syndrome). Mean liver stiffness in those 31 patients was 7.0±2.1kPa (range, 3.1-11.8kPa); 24 were Metavir F0-F1, 7 were F2-F3 while none was F4. Type 1 diabetic patients with abnormal hepatic ultrasound had higher fasting blood glucose, HbA1c and total cholesterol than those with normal findings. Liver stiffness was significantly higher in patients with abnormal liver ultrasound compared with normal sonography. Liver stiffness was positively correlated to HbA1c and ALT. CONCLUSIONS Hepatic abnormalities are prevalent in T1DM and related to poor metabolic control. TE provides a non-invasive method for detection of hepatopathy-induced fibrosis.

Association between thyroid autoimmunity and chronic urticaria in patients versus healthy controls

Introduction There is growing evidence that some cases of chronic idiopathic urticaria are associated with various autoimmune diseases such as thyroid autoimmunity. The association between chronic urticaria (CU) and thyroid disorders has been a subject of controversy. Some reports link CU with hyperthyroidism or hypothyroidism. The frequency of thyroid antibodies in patients with chronic idiopathic urticaria reported in 2009 was 30%, which is higher than that previously reported. Objective This is a case-control study that aimed to detect the presence of markers of thyroid autoimmunity (thyroid autoantibodies with or without underlying abnormal thyroid functions) among a cohort of autologous serum skin test (ASST)-positive patients with CU in comparison with ASST-negative CU patients as well as with healthy controls, and correlating it to the severity of urticaria symptoms. Patients and methods This study was carried out on 80 CU patients attending the Allergy and Immunology Clinic of Ain Shams University Hospitals. CU was diagnosed on the basis of the appearance of continuous recurrent hives for more than 6 weeks. The patients were subdivided into the following groups: group A - 40 CU patients with positive ASST; group B - 40 CU patients with negative ASST. In addition, 40 healthy individuals were included in this study as healthy controls. History and general examination were conducted to all study grouos. Assessment of the Urticaria Activity Score-7 and laboratory investigations including those for complete blood count, erythrocyte sedimentation rate, thyroid function, thyroid Abs, namelyantimicrosomal antibody and antithyroglobulin antibody and total immunoglobulin E (IgE), were done. Results Comparison between the three groups showed that antithyroglobulin antibody was highly statistically significant in group A than in both group B and healthy controls. Moreover, antimicrosomal antibody was also found to be of higher statistical significance in group A than in both group B and healthy controls. Although total IgE had no statistical significance between groups A and B, total IgE was found to be statistically significantly higher in group B than in healthy controls. Level of thyroid stimulating hormone was higher in group A than in controls, and free T3 was lower in group A than in group B. Conclusion We suggest that thyroid diseases have a role in CU, which was confirmed by a higher level of thyroid antibodies in the ASST-positive group than in ASST-negative patients and healthy controls.

The influence of interferon-β supplemented human dendritic cells on BCG immunogenicity

INTRODUCTION Tuberculosis (TB) remains a huge worldwide burden, despite extensive vaccination coverage with the Bacillus Calmette-Guérin (BCG), the only vaccine available against this disease, indicating that BCG-driven immunity is inadequate to protect the human population against TB. This underscore the critical necessitate to develop an improved TB vaccine, based on a better understanding of host-pathogen interactions and immune responses during mycobacterial infection. AIM OF THE WORK To examine whether the exogenous addition of IFN-β could improve dendritic cell (DC) response to Mycobacterium bovis (M. bovis) and to evaluate the effect induced by the infection of human DCs with M. bovis (with and without IFN-β) and Mycobacterium tuberculosis (Mtb) on DC viability as well as to compare the ability of BCG and Mtb to provide DCs with a Th1-polarizing capacity through the assessment of the immunoregulatory cytokines interleukin (IL)-12, IL-10 and interferon-gamma (IFN-γ). METHODS Immature DCs (iDCs) were generated in vitro using peripheral blood monocytes separated by anti-CD14-conjugated microbeads in the presence of granulocyte-macrophage-colony-stimulating factor (GM-CSF) and IL-4, cultured cells were analyzed using flow cytometry, then we tested DC viability after inoculation with M. bovis (with and without IFN-β pretreatment) and Mtb using light microscopic examination and trypan blue exclusion method. Additionally, supernatants from infected-DCs cultures were analyzed for IFN-γ, IL-12 and IL-10 by ELISA. RESULTS The viability of BCG-infected DCs was significantly higher than that of Mtb-infected DCs (61.55% vs 52.10%). BCG-infected DC produced significantly more IL-12 (p = 0.02) and less IL-10 (p = 0.01) compared with Mtb-infected cells. IFN-β-pretreated BCG-infected DCs produced significantly larger amounts of IL-12 than did BCG-infected DCs (p = 0.03) and Mtb-infected cells (p < 0.001). CONCLUSION IFN-β improves DC functions following BCG infection, thus assuming that IFN-β could be used as a vaccine adjuvant.

Evaluation of endothelial protein C receptor in patients with systemic lupus erythematosus: correlation with disease activity and lupus nephritis

Introduction Systemic lupus erythematous (SLE) is a systemic, multifaceted inflammatory disease with clinical manifestations is protean and follows a relapsing and remitting course. Lupus Nephritis (LN) is one of the most frequent and serious manifestation. Endothelial protein C receptor (EPCR) is a transmembrane receptor that is shed into soluble form (sEPCR) in inflammatory status. It is demonstrated as a part of the pathobiology of the SLE disease. Aim of the work To assess correlation of sEPCR level in SLE patients to the disease activity in these patients and to relate sEPCR to LN. Patients and methods Serum level of sEPCR using enzyme-linked immunosorbent assay (ELISA), chemical and immunological markers of SLE were measured in 30 SLE patients and 30 age and sex matched apparently healthy controls. SLE patients were subgrouped into 20 patients without LN and 10 with LN. Disease activity was assessed using SLE Disease Activity Index (SLEDAI). Results A significantly higher sEPCR level was found on comparing SLE patients to controls with statistically highly significant difference (z = 4.8, P < 0.001). Moreover, there was a significantly higher sEPCR level on comparing SLE patients with LN to those without LN with statistically highly significant difference (z = 3.9, P < 0.001). Serum sEPCR had a highly significant positive correlation with SLEDAI in SLE patients (r = 0.66, P < 0.01). Conclusion sEPCR has a possible role in the pathogenesis of SLE and particularly LN diseases, reflecting disease activity in SLE patients.