Iman H. Bassyouni

Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity.

AIM Imbalance of T-helper-cell (TH) subsets (TH1/TH2/TH17) and regulatory T-cells (Tregs) is suggested to contribute to the pathogenesis of Systemic lupus erythematosus (SLE). Therefore, we evaluated their cytokine secretion profile in SLE patients and their possible association with disease activity. METHODS Sixty SLE patients, 24 rheumatoid arthritis (RA) patients and 24 healthy volunteers were included in this study. Demographic, clinical, disease activity and serological data were prospectively assessed. Plasma cytokines levels of TH1 (IL-12, IFN-γ), TH2 (IL-4, IL-6, IL-10), TH17 (IL-17, IL-23) and Treg (IL-10 and TGF-β) were measured by enzyme linked immunosorbent assays (ELISA). RESULTS SLE patients were found to have significantly higher levels of IL-17 (p<0.001), IL-6 (p<0.01), IL-12 (p<0.001) and IL-10 (p<0.05) but comparable levels of IL-23 and IL-4 and slight reduction (but statistically insignificant) of TGF-β levels compared to controls. IL-6, IL-10 and IL-17 were significantly increased (p<0.05) with disease activity. The RA group exhibited significantly higher levels of plasma IL-4 (p<0.01), IL-6 (p<0.05), IL-17 (p<0.001), IL-23 (p<0.01) and TGF-β (p<0.5) and lower IFN-γ (p<0.001) and IL-10 (p<0.01) than those of healthy subjects. CONCLUSION Our study showed a distinct profile of cytokine imbalance in SLE patients. Reduction in IFN-γ (TH1) and TGF-β1 (Treg) with the elevation in IL-6 and IL-17 (TH17) could imply skewing of T-cells toward TH17 cells. Breaking TH17/Treg balance in peripheral blood may play an important role in the development of SLE and could be responsible for an increased pro-inflammatory response especially in the active form of the disease.

CD4+CD25+ regulatory T cells (TREG) in systemic lupus erythematosus (SLE) patients: the possible influence of treatment with corticosteroids.

Systemic Lupus Erythematosus (SLE) is a chronic, systemic autoimmune disease characterized by loss of tolerance to self-antigens. Regulatory T cells (T(REG)) are those CD4+ T cells that constitutively express high levels of CD25 and exhibit powerful suppressive properties. The aim of this work was to quantify CD4+CD25+ (T(REG)) cells and the Mean Fluorescence Index (MFI) of T(REG) in the peripheral blood of patients with SLE and to correlate these findings with their disease activity scores and drug therapy. This study included 24 SLE patients with various disease activity scores (SLEDAI) and 24 healthy age and sex matched controls. Flow cytometry was used to examine the frequency of CD4+CD25+ T cells and the MFI of CD4+CD25+(high) T cells (T(REG)). CD4+CD25+ T cells % and MFI of CD4+CD25+(high) T cells were higher in SLE patients than controls (p value=0.62 and=0.037 respectively) and both CD4+CD25+ T cell % and the MFI of CD4+CD25+(high) T cells showed highly significant correlation with SLEDAI scores (both with a p value<0.001) and were higher in patients taking glucocorticoids than those not on glucocorticoids (p= 0.023, 0.048 respectively). We conclude that the increase in T(REG) cells in our patients may be due to corticosteroid treatment.

Plasma concentrations of growth arrest specific protein 6 and the soluble form of its tyrosine kinase receptor Axl in patients with Systemic lupus erythematosus and Behçets disease

PurposeThe aim of the present study was to investigate plasma concentrations of Gas6 and its soluble tyrosine kinase receptor sAxl in Systemic lupus erythematosus (SLE) and Behçets disease (BD) patients and to correlate those levels with clinical and laboratory manifestations of the diseases.MethodsThe study included 89 female SLE and 49 male BD patients. Twenty-seven age and sex matched healthy volunteers served as controls. All patients were subjected to full clinical examination, laboratory investigations and assessment of disease activity. Plasma concentrations of Gas6 and sAxl were quantified using ELISA technique.ResultsThe level of Gas6 and Axl were significantly altered in the SLE patients (p < 0.001) and in the BD patients (p 0.001 and 0.04 respectively) compared to those of the control. In SLE, the Gas6 was remarkably lower in those with class 1 lupus nephritis and in those with neuropsychiatric manifestations. In the BD patients, the level of Axl was significantly increased in those with neurological disease activity. The number of lymphocytes significantly negatively correlated with the gas6 and Axl levels significantly correlated with the number of neutrophils and negatively with the lymphocytic count in the BD patients.ConclusionThe plasma concentrations of Gas6 and Axl were significantly altered in SLE and BD patients, suggesting that the Axl receptor shedding is an active process affected by and influences Gas6-mediated Axl-signaling in both diseases. Special attention is required in SLE patients with early lupus nephritis and neuropsychiatric manifestations and BD patients presenting with neurological disease activity. The relation with lymphocytes and neutrophils in BD throws light on the role of gas6 and Axl on their known resistance to cell death. Although the mechanisms responsible for the initiation of BD remain to be clarified, the role of the apoptotic process seems critical throughout the disease.

Enthesitis and Related Changes in the Knees in Seronegative Spondyloarthropathies and Skin Psoriasis: Magnetic Resonance Imaging Case-Control Study

Objective. To describe enhanced magnetic resonance imaging (MRI) features and characteristic entheseal changes in the knees in patients with seronegative spondyloarthropathy (SpA). Methods. The 56 patients included 30 with psoriatic arthritis, 5 with ankylosing spondylitis, 5 with reactive arthritis, 5 with ulcerative colitis (UC), 5 with Crohn’s disease, and another 6 with skin psoriasis. Controls were 20 healthy subjects without knee complaints. MRI was performed in all participants, emphasizing entheseal sites. Results. Both knees were studied in 45 (80.3%) patients and one knee in 11 (19.6%). MRI showed evidence of bone marrow edema in 13 (23.2%) patients, cartilaginous erosions in 18 (32.1%), and bone erosions in 9 (16.1%). Enthesitis was found in medial collateral ligaments in 18 (32.1%), lateral collateral ligaments in 8 (14.3%), posterior cruciate ligaments in 3 (5.35%), patellar tendon in 18 (32.1%), biceps femoris insertion in 3 (5.35%), medial patellofemoral ligaments (MPFL) in 5 (8.9%), and lateral patellofemoral ligament in 1 patient (1.8%). In the UC and Crohn’s patients (n = 10), 2 had bone erosions and 5 had enthesitis. In the skin psoriasis group (n = 6), one had bone marrow edema; enthesitis was detected in 5 at the patellar tendon insertion and in one in the MPFL. Entheseal-related changes were absent in the controls. Conclusion. This is the first study showing entheseal-related changes in the knees in patients with inflammatory bowel disease or skin psoriasis without clinical arthritis. Enthesitis of the knee on MRI may be an early finding in SpA.

Clinical significance of anti-cyclic citrullinated peptide antibodies in Egyptian patients with chronic hepatitis C virus genotype IV infection.

Abstract Background: Symmetric polyarthritis associated with hepatitis C virus (HCV) infection frequently displays a clinical picture like rheumatoid arthritis (RA). Antibodies to cyclic citrullinated peptide (CCP) have high specificity for the diagnosis of RA. This study examined the frequency and clinical significance of anti-CCP antibodies in patients with chronic HCV infection, with and without manifestations of joint involvement, compared to RA patients. Methods: Serum anti-CCP antibodies and rheumatoid factor (RF) were evaluated in 30 patients with RA and 47 patients with chronic HCV infection. Of those with HCV infection, 20 patients had chronic HCV infection associated with articular involvement and 27 patients had chronic HCV infection without any articular involvement. Results: Anti-CCP antibody level was positive in 70% of RA patients, 8.5% of HCV-infected patients, and in 20% of HCV patients with articular manifestations. RF was positive in 76% of RA patients and in 60% of HCV patients with articular involvement. Cryoglobulins were found in 29% of HCV-infected patients and in 16% of RA patients. Cryoglobulins were more frequent among HCV patients with articular affection (35%) compared to HCV patients without articular affection (26%). Conclusions: Although anti-CCP antibodies remain a useful diagnostic tool for RA, their interpretation in HCV-infected patients with arthritis should be applied with caution. The possibility that those patients could be prone to develop RA cannot be ruled out. Those patients need careful clinical and radiological follow-up. Further large-scale studies are warranted. Clin Chem Lab Med 2009;47:842–7.

Clinical significance of serum levels of sCD36 in patients with systemic sclerosis: preliminary data

OBJECTIVE To evaluate the clinical significance of anti-angiogenic receptor cluster of differentiation 36 (CD36) in serum of patients with SSc. METHODS We studied 47 SSc patients (28 with lcSSC and 19 with dcSSC) and 38 age- and gender-matched healthy controls. Demographic, clinical, autoantibodies and serological data were prospectively assessed. Peripheral vascular affection was classified into mild, moderate, severe or end-stage based on a vascular severity scale. Soluble CD36 (sCD36) serum levels were measured using ELISA. RESULTS Serum sCD36 levels were significantly higher in patients with SSc compared with healthy controls (P = 0.045). When the patients were divided into clinical subsets, sCD36 was higher in lcSSc than in healthy controls (P = 0.03). Levels of sCD36 were found to be positively correlated with pulmonary artery systolic pressure (PASP) and negatively correlated with percentage diffusing lung capacity for carbon monoxide (DL(CO)). In the multivariate analysis, 50% of the variation of sCD36 levels could be explained by elevated PASP (0.000), telangiectasias (0.026) and increasing vascular severity (P = 0.003). CONCLUSION Serum sCD36 levels were higher in SSc patients (particularly the limited subset) than in healthy controls and were found to be correlated with PASP and vascular severity. We conclude that sCD36 may be a marker for elevated PASP and vascular involvement in SSc. To confirm our results we propose that larger scale, multicentre studies with longer evaluation periods are needed.

Genetic polymorphisms of interleukin 6 and interleukin 10 in Egyptian patients with systemic lupus eythematosus

Systemic lupus erythematosus (SLE) is a complex autoimmune disease. Cytokine gene polymorphisms play an important role in SLE. Thus, this study aimed to investigate the associations between interleukin 6 (IL-6) and interleukin 10 (IL-10) promoter single-nucleotide polymorphisms (SNPs) and their susceptibility to SLE and the implications for plasma levels. We genotyped IL-6-174G/C (rs1800795) using mutagenically separated polymerase chain reaction (MS-PCR) and IL-10-1082G/A (rs1800896) and -819C/T (rs1800871) using sequence specific primer polymerase chain reaction (SSP-PCR) in 100 Egyptian patients and 119 controls. The plasma levels of IL-6 and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). There was significant increase in the frequency of IL-6 (-174) GG genotype (P < 0.05) and G allele (P < 0.01) compared to controls. A significant increase in the distribution of IL-10 (-1082G/A) GG (P < 0.05) and AA (P < 0.05) genotypes and a significant reduction in the frequency of GA genotype (P < 0.05) was found in SLE patients. The mean serum concentration of IL-6 (P < 0.001) and IL-10 (P < 0.001) was significantly elevated in SLE patients compared to healthy controls. There was no  significant association of the most common clinical findings and IL-6 and IL-10 gene polymorphisms in SLE patients. In conclusion, our preliminary study indicated that both GG genotype and G allele of IL-6 (-174G/C) could be considered as risk factors for SLE. In addition, the polymorphisms at IL-10 (-1082 G/G and AA) may play a role in SLE susceptibility in Egyptian patients. Larger prospective studies are needed to confirm our findings.

Serum Concentrations of Cyclooxygenase-2 in Patients with Systemic Sclerosis: Association with Lower Frequency of Pulmonary Fibrosis

Systemic sclerosis (SSc) is a multisystem disease in which interplay between inflammation, autoimmunity and fibrosis appears to play an indispensable role. Owing to the suggested role of cyclooxygenase-2 enzymes (Cox-2) in inflammation and fibrosis, we investigated their serum concentrations in SSc patients and their clinical and laboratory associations. Serum from 49 patients with SSc, 28 of whom had limited cutaneous SSc (lSSc) and 21 had diffuse cutaneous SSc (dSSc) subtypes, and from 27 healthy subjects were assayed for Cox-2 and TNF by enzyme-linked immunosorbent assay (ELISA). Demographic, clinical, autoantibodies and serological data were prospectively assessed. The analysis revealed that patients with lSSc had higher levels of serum Cox-2 than controls. Serum Cox-2 levels were increased in SSc patients with arthritis and digital ulcers; on the contrary, these were diminished in those with associated pulmonary fibrosis. An additional prospective large scale, longitudinal study should be carried out to support these findings and to reveal the mechanistic connections between Cox-2 levels and SSc disease manifestations.

Diminished soluble levels of growth arrest specific protein 6 and tyrosine kinase receptor Axl in patients with rheumatoid arthritis

Growth arrest specific protein 6 (Gas‐6) and its tyrosine kinase receptor Axl plays an important role in apoptosis, and regulation of innate immune response, therefore, we investigated their plasma concentrations in Rheumatoid arthritis (RA) patients and correlated them to clinical, laboratory and radiological parameters of the disease.

Clinical Association of a Soluble Triggering Receptor Expressed on Myeloid Cells-1 (sTREM-1) in Patients with Systemic Lupus Erythematosus.

ABSTRACT A triggering receptor expressed on myeloid cells-1 (TREM-1) is a member of the immunoglobulin superfamily with an established role in innate and adaptive immune response. We aimed to determine the plasma concentrations and clinical association of sTREM-1 in Systemic Lupus Erythematosus (SLE) patients. Plasma from 79 SLE patients and 35 normal healthy subjects were assayed for sTREM-1 and IL-6 levels using Enzyme Linked Immunosorbant Assay (ELISA). The clinical disease characteristics and serological data were prospectively assessed. Disease activity was scored using the SLE disease activity index. We detected significantly higher levels of sTREM-1 in plasma of SLE patients than the healthy control group. We also detected high sTREM-1 levels in subgroups of patients with neuropsychiatric manifestations (NPLE) and patients with the total high disease activity and NPLE activity. In addition, sTREM-l levels were significantly correlated with parameters of disease activity, i.e. SLEDAI score, IL-6, hypoalbuminemia. On the other hand, we did not find significant differences in sTREM-1 levels in relation to age, disease duration, medications, ESR, other organ system involvement, or the presence of anti-dsDNA. Our preliminary data indicated that sTREM-1 levels may be an additional useful marker of disease activity in SLE. It also highlights its importance in patients with NPLE. An additional prospective longitudinal study should be carried out to support these findings.