Evaluation of klotho expression on peripheral blood lymphocytes among hemodialysis patients and its possible contribution to their immuno-compromised status.

Infection is the second most common cause of mortality among end-stage kidney disease (ESKD) patients. Uremic toxins are the main cause of impaired immune response among ESKD patients. Klotho gene, the anti-aging gene, encodes the transmembrane alpha klotho (αKL) protein which acts as an obligate coreceptor for fibroblast growth factor 23 (FGF23). Klotho protein may play a role in immune cell functions, particularly in anti-inflammatory response; however, its role is still incompletely understood. In the present study, we aimed to measure αKL protein expression on peripheral blood lymphocytes (PBLs) among hemodialysis (HD) patients, and we assumed that decreased αKL expression on PBLs may contribute to the impaired immunity among HD patients. This case-control study included 20 ESKD patients on regular hemodialysis for more than 3 months. Their ages ranged from 24 to 69 years. Patients with primary immunodeficiencies, those on systemic immunosuppressive drugs, those with ongoing infections or who had recently recovered from infections, and those with malignancies on active treatment were excluded. A control group of 20 normal subjects of comparable age and gender were also included. We compared αKL protein expression on PBLs by flow cytometry between both groups. Significant reductions in percentages of αKL protein expression on B lymphocytes (CD19), T lymphocytes (CD3), and natural killer cells (CD56) were observed among HD patients compared to controls. We also noticed a significant reduction in the percentages of natural killer cells among HD patients. The present study suggests that decreased αKL expression on PBLs may contribute to the immunocompromised status among HD patients, highlighting the importance of understanding the exact function of αKL protein on immune cells. This may offer a future diagnostic and therapeutic tool to improve the immune response among HD patients.

Neutrophil Respiratory Burst: an immunological indicator of post stroke infection.

Stroke is long known to be followed by a series of immunosuppressive events, and infections might be a cause of death after an acute insult of stroke. The aim of our work was to assess the percentage of neutrophils showing spontaneous oxidative burst in patients with acute ischemic stroke. The study included 30 patients with acute cerebral infarction subjected to the following: magnetic resonance imaging of the brain immediately on admission, and blood sampling on day one of admission (baseline) and after 3 days of admission. Blood samples were used for the assessment of: differential leucocyte count and percentage of neutrophils showing spontaneous oxidative burst, performed by flow cytometry. Thirty age and gender matched controls were also recruited. Neutrophil respiratory burst percentage was significantly lower in stroke patients in comparison to controls (P < 0.001), and stroke patients had significantly lower neutrophil respiratory burst percent on day 3 of admission compared to the baseline (P < 0.001). Stroke-induced immune alterations including impairment of the first-line defense performed by neutrophils against bacteria. The hypothesis that these changes enhance susceptibility to acquired infections is supported by our observation that oxidative burst in neutrophils was more impaired in patients with stroke who exhibited subsequent stroke-associated infections.

Toll like receptor 4 (TLR4) expression on peripheral blood mononuclear cells (PBMCs) among patients with Graves disease.

Graves disease (GD) is a multifactorial disease due to multiple environmental and genetic factors as well as immune malfunction. Human Toll-like receptors (TLRs) play a key role in activating innate and adaptive immune cells. Their role in modulating immunity also interferes with the mechanisms that maintain tolerance in the host. Thus, expression or activation of TLRs can contribute to the loss of tolerance by a lot of mechanisms. In order to confirm the importance of TLR4 in the pathogenesis of GD, this study intended to measure TLR4 expression on peripheral blood mononuclear cells in GD patients before and after control of disease with Carbimazole as compared to a group of normal controls. We conducted a case-control study on 36 patients with newly diagnosed Graves disease and 36 individuals as the control group. Patients were recruited from the endocrinology outpatient clinic, Ain-Shams University Hospitals and were followed up till achieving the euthyroid state (for a minimum period of 6 months). In GD patients at baseline, the mean monocyte percentage was 4.46%, and the mean TLR4 on monocytes 90.91%. After achieving euthyroid state, the mean monocyte was 6.16%, and the mean TLR4 on monocytes 72.30%. In the control group, the mean monocyte was 3.88%, and the mean TLR4 on monocytes 66.30%. Results indicated significant differences in expression of TLR4 between GD patients before treatment, after achieving euthyroid state and in the control group (p< 0.0001). In conclusion, the present study showed a higher expression of TLR4 on monocytes among newly diagnosed GD patients in comparison to normal individuals. TLR4 expression on monocytes decreased significantly among GD patients after treatment.

Low frequency of CD4+CD25highFOXP3+ T regulatory cells in Egyptian patients with chronic spontaneous urticaria.

Chronic urticaria is a prevalent disabling dermatological disease. About 90%, are considered idiopathic and referred to as chronic spontaneous urticaria (CSU), and nearly half of them are likely to have autoimmune mechanisms. Regulatory T cells play a substantial role to prevent autoimmune diseases. Subsets of Tregs expressing the CD4+CD25high and forkhead-box-P3 (FOXP3) transcription factor, crucial for their development and function, are best characterized in maintenance of self-tolerance. The objective of this study was the analysis of peripheral CD4+CD25highFOXP3+(T regs) frequency in chronic spontaneous urticaria; and its possible association with autologous serum skin test (ASST). Fifty chronic spontaneous urticaria patients (25 with positive ASST and 25 with negative ASST) and 20 healthy controls were enrolled in this study. The frequency of CD4+CD25highFOXP3+ (T regs) was analyzed by flow cytometry. A Significant decrease in peripheral blood CD4+CD25highFOXP3+ T regs% was detected in CSU patients in comparison to healthy individuals (median [IQR], 1.47% [0.71-3.12] vs 1.79% [1.15-4.00]; P = 0.05). When ASST positive patients were compared with ASST negative patients, no significant difference was found in percentage of T regs, (P=0.112). In conclusion our data provided further insights into CSU pathogenesis. Reduced frequency of CD4+CD25highFOXP3+(Tregs) in patients with urticaria, support the notion that CSU is an immune mediated disease and may help researchers to develop a novel immunotherapy strategy.

The interplay between tumor necrosis factor alpha, disease activity, and depressive symptoms among Egyptian female patients with systemic lupus erythematosus.

One of the most remarkable presentations of systemic lupus erythematosus (SLE) is depression. Our aim was to elucidate the potential relationship between disease activity, depressive symptoms, and tumor necrosis factor alpha (TNF-α) in patients with SLE. Sixty female patients with SLE and thirty comparable healthy controls were recruited. According to systemic lupus erythematosus disease activity index, patients were subdivided into two similar groups; active and inactive. Complete clinical and laboratory assessments were done to authenticate the diagnosis of SLE and outline its activity. All participants were assessed using the Beck depression Inventory (BDI) to diagnose and determine the severity of depressive symptoms. TNF-α level was assessed using Enzyme linked immunosorbent assay technique. Using BDI, patients with SLE activity showed higher prevalence of depression 19 (63.3%) compared to those with inactive SLE and control groups (P < 0.001). TNF-α level was markedly elevated amongst patients with active SLE in comparison to inactive and control groups (P <0.001). TNF-α differentiated SLE patients into with and without depression at cut-off value (>360 ng/l) (AUC = 0.726; P=0.0008; 95% CI 1.3-2.7). Multivariable regression analysis for prediction of depression revealed that TNF-α was the only independent predictor of depression (P= 0.011). In conclusion, patients with increased SLE activity are more prone to depression especially, moderate to severe degree. TNF-α level could be of significance in predilection of depression and SLE activity in patients with SLE. Hence, future studies are essential to test the treatment modalities targeting TNF-α in those patients.

Skin Prick Test Reactivity to Aeroallergens among Egyptian Patients with Isolated Allergic Conjunctival Disease.

Allergic conjunctival disease (ACD) is a type of ocular allergy, which includes seasonal allergic conjunctivitis (SAC), perennial allergic conjunctivitis (PAC), and vernal keratoconjunctivitis (VKC). Little is known about the pattern of sensitization or prevalent aeroallergens among patients with isolated ACD in Egypt We aimed to evaluate the prevalence of skin prick test positivity to common aeroallergens among Egyptian patients with isolated allergic conjunctival disease. The study included 75 patients with isolated ACD recruited from a tertiary Egyptian outpatient clinic. Skin prick test (SPT) was performed for all patients with common aeroallergens. Total serum immunoglobulin E (IgE) was measured by ELISA. A positive SPT reaction was present among 32 patients (42.7%). The most prevalent aeroallergens among all patients were mites and pollens (12% respectively), followed by grass (8%) and hay dust (6.7%). Eight patients (10.7%) had SAC, 19 patients (25.3%) had PAC, and 48 patients (64%) had VKC. Prevalence of SPT positivity to indoor allergens was significantly more common among PAC (52.6%) than among SAC (25%) and VKC (16.7%), P= 0.011. Outdoor allergen sensitization did not differ significantly between the 3 subgroups, P= 0.614. Elevated IgE levels were observed among 62.5%, 73.7% and 66.7% of patients with SAC, PAC and VKC, respectively, with no statistically significant difference between them, P= 0.806. In conclusion aeroallergen sensitization is common among Egyptian patients with isolated ACD. Accordingly, SPT should be included in the diagnostic workup of these patients.

Skin test reactivity to female sex hormones in women with primary unexplained recurrent pregnancy loss.

The objective was to examine the hypothesis that primary unexplained recurrent pregnancy loss might be associated with an inappropriate immunologically mediated response to progesterone and/or estrogen. This prospective study included 47 women with two or more documented consecutive early pregnancy losses of unknown etiology, and no previous history of deliveries. Intradermal skin testing was performed in the luteal phase of the cycle (days 16-20) using estradiol benzoate, progesterone, and a placebo of refined sesame oil. Immediate (20 min) and late (24h and 1 week) skin test readings for all cases were compared with those of 12 parous women of comparable age with no history of spontaneous miscarriages, premenstrual disorders, pregnancy, or sex hormone-related allergic or autoimmune diseases. Main outcome measure was skin test reactivity to estradiol and/or progesterone. Immediate skin test reactivity to both hormones was observed among half of the cases at 20 min. A papule after 24h, which persisted for up to 1 week, was observed among 32 (68.1%) and 34 (72.3%) cases at the sites of estrogen and progesterone injection, respectively. 55.3% of cases had combined skin test reactivity to both estradiol and progesterone at 1 week. All women in the control group showed absence of skin test reactivity for both estradiol and progesterone at 20 min, 24h, and 1 week. None of the subjects in either group showed skin test reactivity to placebo. There is an association between primary unexplained recurrent pregnancy loss and skin test reactivity to female sex hormones.

Study of folate status among Egyptian asthmatics.

OBJECTIVE: To compare serum folate levels between atopic asthmatics, non-atopic asthmatics, and healthy controls. METHODS: This case-control study included 60 asthmatics with at least one positive skin prick test (SPT) reaction (atopic asthma group), 60 asthmatics with negative SPT reactions (non-atopic asthma group), and 60 healthy controls with no history of asthma or other allergic diseases, and with negative SPT reactions. Serum folate and total IgE levels were measured in all subjects. In addition, lung functions were assessed by spirometry. RESULTS: Serum folate levels were significantly lower among the atopic asthma group [9.1 (4.9, 12.1) ng/mL] as compared to the non-atopic asthma group [11.3 (7.5, 14.8) ng/mL] and the control group [12.0 (8.3, 15.1) ng/mL], p= 0.001. Among atopic asthmatics, serum folate levels were inversely correlated with total serum IgE levels (r=-0.483, p<0.001), and the number of positive SPT reactions (r=-0.442, p<0.001). Atopic asthmatics with a total serum IgE ≤200 IU/mL had significantly higher levels of serum folate than those with a total serum IgE >200 IU/mL. Regression analysis showed that higher folate levels independently predicted lower total serum IgE levels. Folate was not found to be an independent predictor of asthma. No association was observed between serum folate levels and values of forced expiratory volume in 1s. CONCLUSION: Among asthmatics, serum folate levels are significantly lower among atopics, and correlate inversely with the degree of atopy.

OX40/OX40L in systemic lupus erythematosus: association with disease activity and lupus nephritis.

BACKGROUND AND OBJECTIVES: OX40-OX40L interaction is implicated in the pathogenesis of systemic lupus erythematosus (SLE). We evaluated the role of OX40/OX40L as markers of disease activity and nephritis in SLE patients. DESIGN AND SETTING: Case-control study conducted in 2009 on SLE patients attending the outpatient clinics of Ain Shams University Hospital, Egypt. PATIENTS AND METHODS: We assessed the percentage of CD4+ T-lymphocytes expressing OX40 by flowcytometry, and serum OX40 ligand (OX40L) levels in 40 patients with SLE (20 with lupus nephritis and 20 without) and in 20 healthy controls. Disease activity was assessed by the University of Toronto SLE disease activity index (SLEDAI). RESULTS: The percentage of CD4+ T-lymphocytes expressing OX40 was significantly higher in SLE patients than in controls, and in patients with lupus nephritis than in those without. OX40 expression correlated positively with both serum creatinine levels and SLEDAI. OX40 expression was the highest in patients with class V lupus nephritis and lowest in class II. Serum OX40L levels were significantly higher in SLE patients than in controls, and in patients with nephritis than in those without. Serum OX40L levels correlated with serum creatinine levels but not with SLEDAI. OX40 expression on CD4+ T-cells had a higher sensitivity and specificity in diagnosing lupus nephritis than both OX40L and anti-double-stranded DNA levels. CONCLUSION: OX40-OX40L interaction plays a role in the pathogenesis of SLE. The expression of OX40 on CD4+ T-lymphocytes and the serum level of OX40L may act as markers of lupus nephritis. Measurements of percentages of CD4+ T-lymphocytes expressing OX40 may serve as an indicator of disease activity in SLE.