Performance of two multiplex flow cytometric assays for antibody detection in Egyptian patients.

Background: Autoantibodies are vital biomarkers for the diagnosis, assessment and prognostic determination of various autoimmune disorders. Objective: This study aimed to evaluate the performance of the two AtheNA Multi-Lyte® systems for the detection of various autoantibodies. Methods: A total of 105 systemic lupus erythematosus patients, 35 patients with other autoimmune diseases (diseased controls), and 30 healthy volunteers (healthy controls) at Zagazig University Hospitals, Zagazig city, Al Sharqia governorate were tested for anti-double-stranded DNA (anti-dsDNA) antibodies using indirect immunofluorescence (IIF) and the AtheNA Multi-Lyte® anti-nuclear antibodies-II system between May 2020 and April 2022. Seventy-five patients with clinically suspected autoimmune vasculitis (AIV) and 25 healthy volunteers were also tested for anti-myeloperoxidase and anti-proteinase 3 antibodies using IIF, the AtheNA Multi-Lyte® AIV system, and enzyme-linked immunosorbent assay (ELISA). Results: The AtheNA anti-dsDNA test (98.5%) was more specific than IIF (96.9%) for diagnosing systemic lupus erythematosus, but both tests had the same sensitivity (38.1%). Combining both methods increased sensitivity to 47.6%, while increasing the cut-off of the AtheNA anti-dsDNA test to 134 international units/mL increased specificity to 100%. The AtheNA Multi-Lyte AIV system exhibited substantial agreement with IIF regarding anti-myeloperoxidase testing (κ = 0.65) and almost perfect agreement with ELISA (κ = 0.85). The AtheNA Multi-Lyte® AIV system exhibited perfect agreement with IIF (κ = 1) and substantial agreement with ELISA for anti-proteinase 3 testing (κ = 0.63). Conclusion: AtheNA Multi-Lyte® systems appear to be reliable for anti-dsDNA, anti-myeloperoxidase, and anti-proteinase 3 screening and may be an optimal choice for monitoring anti-dsDNA levels. What this study adds: It is necessary to evaluate various autoantibodies detection assays to increase both sensitivity and specificity of autoimmune diseases diagnostic approaches. AtheNA Multi-Lyte® systems appear to be reliable for anti-dsDNA, anti-myeloperoxidase, and anti-proteinase 3 screening and may be an optimal choice for monitoring anti-dsDNA levels.

HLA-DRB1, IRF5, and CD28 gene polymorphisms in Egyptian patients with rheumatoid arthritis: susceptibility and disease activity.

This study was established to assess the effects of IRF5 rs10488631 and CD28 rs1980422 single-nucleotide polymorphisms (SNPs) and HLA-DRB1 shared epitope (SE) allele on the prognosis and disease activity of rheumatoid arthritis (RA) patients. A total of 150 RA patients and 150 healthy controls were genotyped for the selected SNPs by real-time PCR. HLA-DRB1 SE was determined using LAB Type SSO Class II DRB1 typing. Our results suggest that HLA-DRB1, CD28, and IRF5 significantly discriminated (p < 0.001) RA patients and healthy controls (OR of single HLA-DRB1 SE allele = 2.431, CI = 1.467-4.027, OR of two SE alleles = 11.152, CI = 2.479-50.159), (OR of CD28 risk allele C = 2.794, 95% CI = 1.973-3.956) and (OR of IRF5 risk allele C = 4.925, CI = 3.26-7.439). Rheumatoid factor (RF) seropositivity was associated with HLA-DRB1 SE (p < 0.001) and IRF5 risk allele (p < 0.001). ACPA was significantly associated only with IRF5 risk allele (p < 0.001). A better response to methotrexate therapy was found in HLA-DRB1 SE non-carriers, and CD28 TT patients. This study demonstrated associations of HLA-DRB1 SE, CD28, and IRF5 with the risk of RA. HLA-DRB1 SE and CD28 rs1980422 can be used as predictors of methotrexate therapy response.

Serum angiopoietin-2 level as a novel biomarker in atopic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is a chronic, pruritic inflammatory skin disease. Angiopoietin is an intrinsic mediator induced by T helper 2 cytokines and has been reported it to play a critical role in the pathogenesis of AD. OBJECTIVES: To determine the relationship between circulating angiopoietin-2 (Ang-2) in AD patients and to determine its correlation with the severity and chronicity of the disease. METHODS: The present study included 38 AD patients and 38 healthy controls, age- and sex-matched. We collected 3 mm of venous blood from each subject. Enzyme-linked immunosorbent assay (ELISA) determined serum angiopoietin. We determined the correlation between serum Ang-2 level, duration of disease, severity, type of atopy, and scoring atopic dermatitis (SCORAD) score. RESULTS: There were highly significant differences between patients and controls with regard to serum Ang-2 levels (P < 0.001). There were significant differences between severe AD patients with serum Ang-2 levels with an increasing level of degree severity (P < 0.001). There were significant differences in serum Ang-2 levels among widespread type (P < 0.03). There was a positive significant correlation between Ang-2 levels and both duration and SCORAD score of the patient group. CONCLUSION: Serum Ang-2 levels may correlate with disease severity, chronicity in patients with AD, and serve as a potential biomarker of the disease severity.

Association between CD4+, CD25+, FOXP3+ Regulatory T-cells and Cardiovascular Complications in Diabetic Patients Type 1.

T regulatory cells (Tregs) are a cornerstone regulator for immune responses and inflammatory reactions. Abnormal number or function of Tregs causes deranged immune response that increases the autoimmune disorders and inflammatory conditions. Type 1 diabetes mellitus is an autoimmune disease associated with many complications, of which, Cardiovascular complications are fundamental and responsible for profound morbidity and mortality. Understanding the immunopathogenesis of these disorders allows early diagnosis and better management by innovating new therapeutic targets. In this study, we aimed to detect the association between CD4+CD8+FOX3+ Tregs, T1DM, and associated cardiovascular complications. The study included 144 individuals divided into three groups, group 1 included 48 patients suffering from T1DM without cardiovascular complications, group II: included 48 type T1DM patients with cardiovascular complications. Group III: included 48 healthy control subjects. For all participants, markers for inflammation, and cardiovascular involvement were assessed. The percentage of CD4+ CD25+ FOXP3+ Regulatory T- cells (Tregs) was measured by flow cytometry using peripheral blood samples. The level of Treg was lowest in group II and highest in group III, the difference was highly significant P < 0.001. Treg in group I significantly correlated with age (r= 0.58, P=0.004), CK-mb (r= 0.61, P=0.04) and LDL (r= -.61, P=0.4). While in group II, it correlated with triglyceride level, (r= 0.65 and a P =- 0.02). In conclusion, Lower levels of Tregs are associated with cardiovascular complications in TIDM patients.

Chitinase-3-like protein 1 (YKL-40): novel biomarker of lichen planus.

BACKGROUND: Lichen planus (LP) is an inflammatory skin disorder of unknown etiology. Chitinase-3-like protein 1 (YKL-40) is a multifunctional proinflammatory protein, which has an important role not only in immunity and inflammation, but also in the regulation of cell proliferation, differentiation, and apoptosis. AIM OF THE WORK: The present study was designed to investigate the serum level of YKL-40 in patients with different clinical presentations of LP in comparison to healthy participants in order to study its possible role in the pathogenesis of LP. PATIENTS AND METHODS: A total of 60 consecutive subjects were enrolled in this prospective case-control study including 30 patients with different clinical variants of LP (16 male and 14 female patients) in addition to 30 healthy controls matched for age and gender. The serum level of serum YKL-40 was evaluated by means of ELISA method in both patients and controls. RESULTS: The serum YKL-40 level was significantly higher (P < 0.001) in patients than in controls. Also, significantly higher levels were found in patients with oral type than in patients with cutaneous type who do not have oral lesions (P < 0.05). The sensitivity and specificity of serum YKL-40 were 62.5 and 81.8%, respectively, (cut off value of 151 pg/ml) for the diagnosis of oral lesions in LP. CONCLUSION: The findings in our study support the role of YKL-40 in the pathogenic process of LP and may help in treatment of lichen planus.