Hyperleptinemia as a Marker of Various Phenotypes of Obesity and Overweight in Women with Rheumatoid Arthritis and Systemic Lupus Erythematosus.

The objective of the study was to identify different phenotypes of overweight in women with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) based on body mass index (BMI) and serum leptin levels, as well as to determine the frequencies of various metabolic disorders, hypertension, and cardiovascular complications (CVCs) in individual phenotypes. The study included 50 women with RA and 46 with SLE aged 18 to 65 years without a history of diabetes and fasting hyperglycemia. In all patients, the concentration of leptin was determined by ELISA, the concentration of insulin was determined by electrochemiluminescence analysis, and the HOMA-IR index was calculated. Hyperleptinemia was diagnosed at leptin concentrations > 11.1 ng/mL; insulin resistance (IR), at HOMA-IR values ≥ 2.77. Three main phenotypes of overweight were distinguished: "classic" (BMI ≥ 25 kg/m2 + hyperleptinemia), "healthy" (BMI ≥ 25 kg/m2, without hyperleptinemia), "hidden" or "latent" (BMI < 25 kg/m2 + hyperleptinemia), as well as "normal weight" (BMI < 25 kg/m2, without hyperleptinemia). Patients with RA and SLE were similar in age (p = 0.4), disease duration (p = 0.2) and BMI (p = 0.5). Hyperleptinemia was found in 46% of women with RA and in 74% of women with SLE (p = 0.005), and IR was found in 10 and 22% of patients, respectively (p = 0.2). The "classic" phenotype of overweight was diagnosed in 30%, "healthy" in 8%, and "hidden" in 16% of cases with RA and in 44%, 0%, and 30% of cases with SLE, respectively. IR was found in 3% and hypertension in 6% of patients with "normal weight." With the "classic" phenotype, IR (29%) and hypertension (66%) were more common than with "normal weight" (p < 0.01 in all cases); with the "hidden" phenotype, significant differences were obtained only in hypertension frequency (45%; p = 0.0012), but not IR (18%). Three out of four women with a history of cardiovascular complications suffered from "classic" overweight, and one patient had a "normal weight." In women with SLE up to 65 years of age, the frequency of hyperleptinemia, but not IR, is higher than in patients with RA. In both diseases, the "classic" overweight phenotype is most common. In RA, a "hidden" phenotype was detected less often than in SLE, at the same time, a "healthy" phenotype is not characteristic of SLE. The frequency of metabolic disorders and hypertension is low with the "normal weight" and "healthy" phenotype, high with the "classic" phenotype, and intermediate with the "hidden" phenotype.

Clinical Significance of Antibodies to DFS70 in Immunoinflammatory Rheumatic Diseases.

The relevance of the problem of immunoinflammatory rheumatic diseases (IIRD) for modern medicine is determined by their high prevalence in the population, the difficulty of early diagnosis, the rapid development of disability and poor life prognosis. Recent data on the significance of anti-DFS70 have opened up new possibilities for optimizing the step-by-step diagnosis of IIRD. The detection of these antibodies can help in the interpretation of a positive result for antinuclear antibodies (ANA) by indirect immunofluorescence assay on HEp-2 cells (IIFA-HEp-2) in the absence of autoantibodies specific for IIRD. Detection of anti-DFS70 in antinuclear factor (ANF) seropositive patients without clinical and/or serological markers characteristic of a certain disease from the IIRD group can be considered as a potential marker that excludes this group of diseases.

[Clinical value of multiplex immune assay of antinuclear antibodies in systemic lupus erythematosus.]

A promising trend in the diagnosis of systemic autoimmune diseases is the multiplex immune assay (MIA) of autoantibodies and other laboratory biomarkers using microchips. The aim of the work was to study the diagnostic and prognostic significance of MIA antinuclear antibody (ANA) profiles in systemic lupus erythematosus (SLE). 94 patients with SLE, 70 patients with other rheumatic diseases and 30 healthy donors were examined. ANA (antibodies to doublestranded - dsDNA, Sm, SS-A/Ro, SS-B/La antigens, nucleosomes, ribosomal protein P-RibP and ribonucleoprotein - RNP-70) were determined in the serum by MIA using the xMAP technology. In MIA, antibodies to dsDNA, Sm and RibP have a high diagnostic specificity (Sp) (95.0-99.0%) and a likelihood ratio of positive results (LR+) (9.67-15.0), i.e. are the most "useful" diagnostic tests, and antibodies to RNP-70, SS-A/Ro and nucleosomes are classified as "useful" tests for the diagnosis of SLE (Sp: 84.0-95.0%, LR+> 2.0). Determination of profiles from 3 or more antigen-specific ANA by MIA increases the Sp method to 98.0-100%, and the LR+ - to the maximum values. Profiles from 7 subpopulations of ANA (antibodies to dsDNA, Sm, RibP, SS-A/Ro,SS-B/La, nucleosomes and RNP-70, 57.9%, 71.9%, 82.5%, 61.4 %, 84.2%, 50.9%, 84.2%) were found in the chronic variant of SLE. In the acute course of the disease, 4 subpopulations of ANA are simultaneously detected (antibodies to dsDNA, Sm, SS-A/Ro and nucleosomes, 77.3%, 45.5%, 40.9% and 72.7%); in subacute course there are 2 subpopulations of ANA (antibodies to dsDNA and nucleosomes, 53.3% and 46.7%). The activity index of SLEDAI-2K positively correlates with the concentration of antibodies to dsDNA (r = 0.55, p < 0.05), nucleosomes (r = 0.65, p < 0.05), RibP (r = 0.32; p < 0.05) and Sm (r = 0.36, p < 0.05) in the blood. There was no reliable relationship between the production of varieties of ANA and the index of organ damage. Mucocutaneous disorders, lupus-nephritis and neurolupus were most often associated with the detection of antibodies to dsDNA (53.2-64.0%), nucleosomes (55.3-66.0%), SS-A/Ro (38.0-40.4%) and Sm (27.8-36.2%). MIA of ANA profiles is an important tool for implementing a personalized approach to diagnosis, evaluation of activity, course and clinical and immunologic subtypes of SLE.

[THE CLINICAL INFORMATIVENESS OF AUTOMATED METHODS OF SCREENING DETECTION OF ANTI-NUCLEAR ANTIBODIES USING INDIRECT REACTION OF IMMUNE FLUORESCENCE, ENZYME-LINKED IMMUNOSORBENT ASSAY AND MULTIPLEX XMAP TECHNOLOGY UNDER SYSTEMIC LUPUS ERYTHEMATOSUS].

Thew antinuclear antibodies (ANA) consist heterogeneous group of auto antibodies reacting with various components of nucleus and cytoplasm. The ANA is a main serological marker of systemic lupus erythematosus (SLE). The implementation in clinical practice of new highly productive techniques of immune analysis using automated systems sets up prerequisites for standardization and amelioration of reproducibility of detection of ANA. The study was carried out to compare diagnostic significance of automated techniques of screening detection of ANA (indirect immunofluorescence test on cells HEp-2 (IIFT-HEp-2)), enzyme-linked immunosorbent assay (ELISA) and multi-complex immune analysis (MIA, using suspension technology xMAP) in serum of patients with SLE. The serums from 94 patients with SLE were analyzed. The comparison group included 70 patients with other rheumatic diseases. The control group consisted of 30 healthy donors. The screening detection of ANA using technique IIFT-HEp-2 was implemented on automated platform AKLIDES, ELISA - on automated analyzer ALEGRIA and MIA on automated analyzer BioPlex 2200. The technique IIFT-HEp-2 demonstrated the most high diagnostic sensitivity as compared with ELISA and MIA- BioPlex 2200 (96.8%; 79.8% and 82.9% correspondingly). The general diagnostic specificity of detection of ANA using technique IIFT-HEp-2 was lower than in case of ELISA and MIO-BioPlex 2200 (40%, 70% and 57% correspondingly). In the group of healthy donors the lowest diagnostic specificity was observed in ANA screening analysis using MIA-BioPlex 2200 (80%) while in case of applying IIFT-HEp-2 and ELISA indices of diagnostic specificity made up 93.3% and 96.7% correspondingly. The ANA analysis of mix of 26 nuclear antigens using ELISA technique was a reliable laboratory test for diagnostic of SLE (likelihood ratio of positive result - 2.66). By the level of likelihood ratio of negative result of the IIFT-HEp-2 technique was more informative test for exclusion of diagnosis of SLE than techniques of ELISA and MIA-BioPlex 2200 (0.08; 0.29 and 0.3 correspondingly). The detection of ANA using technique of is the most preferable primary screening test for diagnostic of SLE. The ELISA of antibodies to mix of nuclear antigens and MIA on the basis of xMAP technology are less preferable screening tests for diagnostic of SLE as compared with IIFT-HEp-2 because of false-negative results in 20% and 17% of cases correspondingly. ELISA and MIA are to applied as confirmatory screening tests permitting to detect antigen-specific ANA in patients with SLE with positive results of IIFT-HEp-2.

[THE IDENTIFICATION OF SUB-POPULATIONS OF B-LYMPHOCYTES OF PERIPHERAL BLOOD USING TECHNIQUE OF FLOW CYTOFLUOROMETRY IN HEALTHY INDIVIDUALS AND PATIENTS WITH RHEUMATOID DISEASES].

The study was carried out to apply technique of flow cytofluorometry for immunofenotyping of sub-populations of B-cells of peripheral blood in healthy persons and patients with rheumatoid diseases. The samples included 27 healthy donors, 16 patients with rheumatoid arthritis and 9 patients with systemic lupus erythematosus. The peripheral blood of all participants was analyzed for relative number of CD19+B-cells, total population of memory B-cells (CD19+CD27+); unswitched (CD19+IgD+CD27+) and switched (CD19+IgD- CD27+) memory B-cells, naive (CD19+IgD+CD27-) and transitory (CD19+IgD+CD10+CD38++CD27-) B-cells, plasmablasts (CD19+CD38+++IgD-CD27+CD20+) and long-lived plasmatic cells (CD19+CD138+). The sub-populations of B-cells were identified by technique of multicolor flow cytofluorometry using panel of monoclonal antibodies to surface membrane markers of B-lymphocytes. In normal state, relative number (median-Me; interquartile range 25-75 percentiles) of CD19+B-lymphocytes amounted to 9.1 (6.6-11.6)%; CD19+CD27+memory B-cells - 2.2 (1.6-3.3)%; unswitched and switched memory B-cells - 10 (6.4-12.7)% and 17.7 (14.9-27.0)%; naive B-cells - 65.8 (55.1-73.4)%; transitory B-cells - 0.1 (0.1-0.3)%; plasmablasts - 7.0 (5.0-9.4)%. The long-lived plasmatic cells in peripheral blood were absent. In patients with rheumatoid arthritis percentage of content of total population of memory B-cells were lower than in donors (1.6; 1.00-2.3%; p = 0.038). Under systemic lupus erythematosus was detected decreasing of number of naive B-cells (40.2; 19.7-58.2) and increasing of level of switched memory B-cells (34.2; 21.0-52.7) as compared with donors (p = 0.003 in both cases). The study established no reliable differences between healthy donors and patients with rheumatoid arthritis and systemic lupus erythematosus in the rest of sub-populations of B-lymphocytes. The developed technique of multi-parametric flow cytofluorometry can be recommended for studying homeostasis of B-cells of peripheral blood in healthy persons and patients with auto-immune rheumatoid diseases and also for evaluating and forecasting effectiveness of anti-B-cells therapy.

[Soluble CD40 ligand in systemic lupus erythematosus: link with atherosclerotic vascular affection].

AIM: To evaluate clinical implications of pCD40L as a marker of atherosclerotic vascular affection in systemic lupus erythematosus (SLE). MATERIAL AND METHODS: The examination of 132 females (mean age 35 years, SLE duration 96 months) assessed classic factors of atherosclerosis risk (AR), total coronary risk (TCR), detected subclinical atherosclerosis with ultrasonic scanning of the carotid arteries. Serum level of pCD40L was measured with enzyme immunoassay (EL4). RESULTS: Concentration of pCD40L in SLE patients was higher than in the control group (7.1 +/- 4.9 and 5.8 +/- 3.1 ng/ml, respectively) but the difference was insignificant (p > 0.05). Upper limit of normal value (M + 2SD) for pCD40L was 12 ng/ml. Elevation of pCD40L level was seen in 21% of SLE patients. Clinical manifestations of atherosclerosis risk were seen in 13% SLE patients, subclinical in 19 and 17% (increased intima-media thickness and atherosclerotic plaques, respectively). No correlations were found between pCD40L level and atherosclerotic symptoms. The pCD40L level was higher in SLE patients with the plaques than in those without them (p = 0.005). A positive correlation exists between a pCD40L concentration and maximal intima-media thickness (r = 0.2; p = 0.02), total cholersterol (r = 0.3; p = 0.002), LDLP cholesterol (r = 0.3; p = 0.004), LDLP cholesterol (r = 0.2; p = 0.04) and age of the patients (r = 0.2; p = 0.03). In patients with TCR > 20% a pCD40L was significantly higher than in patients with TCR < 20% (p = 0.01). CONCLUSION: Elevated pCD40L level is a marker of atherosclerotic affection of the vessels, has an important clinical role for predicting risk of cardiovascular diseases in SLE and elicidation of the role of activation of cell immunity in development of atherosclerosis in this disease.

[Interleukin-18 in systemic lupus erythematosus: link with clinical symptoms and vascular atherosclerosis].

AIM: To examine correlation between IL-18 concentration, SLE manifestations and atherosclerotic vascular affection. MATERIAL AND METHODS: We examined 162 SLE female patients (age 26-43 years, mean age 35 years, duration of SLE 28-204 months, mean duration 96.0 months) and 64 healthy controls (women aged 30.0-45.0 years, mean age 35.5 years). We analysed correlation between IL-18 concentration, SLE symptoms, classic risk factors for cardiovascular diseases, subclinical and clinical signs of atherosclerosis. RESULTS: IL-18 in blood serum of SLE patients was much higher than in the control group (p < 0.00001). A positive correlation exists between IL-concentration and the disease activity by scale SLEDAI-2K, the level of antibodies to IgG cardiolipin, ESR, SLE, titers of antibodies to double-chain DNA, concentration of creatinin, urine, glucocorticoids dose taken by the patients at examination. In SLE patients with stomatitis, hematological, immunological disorders, a positive test for antinuclear factor, IL-18 was much higher than in patients without the above signs. No significant correlation was seen between IL-18 concentration, subclinical and clinical signs of atherosclerosis, Of risk factors, there was a negative correlation between total cholesterol concentration, LDLP and HDLP cholesterol. CONCLUSION: In SLE patients a high IL-18 level reflects activity of the basic disease and is not related with vascular atherosclerosis.

[Clinical significance of heart rate variability in patients with systemic lupus erythematosus].

AIM: To evaluate clinical significance of heart rate variability (HRV) in patients with systemic lupus erythematosus (SLE). MATERIAL AND METHODS: HRV was investigated by means of time-domain analysis of 24 hour ambulatory ECG of 122 SLE patients under 55 years of age and 32 age-matched healthy controls. In addition to clinical manifestations and activity of SLE, we assessed the presence of basic conventional cardiovascular risk factors (hypertension, smoking, body mass index, dyslipidemia), performed common carotid duplex scanning with measurement of intima-medial thickness (IMT). Inflammatory markers (ESR, CRP, IL-6) were assessed in all the patients. RESULTS: Significantly lower HRV and the trend to tachycardia were detected in SLE patients when compared to the control group. There was a significant positive correlation between HRV and a cumulative dose of cyclophosphamide, a high density lipoprotein cholesterol level, a negative correlation between HRV and cumulative dose of azathioprine, standard risk factors (hypertension, smoking, body mass index, triglyceride level), markers of inflammation (ESR, CRP, IL-6) and IMT. CONCLUSION: Measurement of HRV in combination with routine cardiovascular risk factors and level of inflammatory markers can be used for identification of subjects at risk for faster progression of atherosclerosis in SLE patients.

[The significance of cardiovascular risk factors and C-reactive protein to the development of atherosclerosis in women with systemic lupus erythematosus].

Compared with general population, women suffering from systemic lupus erythematosus (SLE) have signs of coronary artery disease (CAD) five to eight times more often, especially in young age. Early development of atherosclerosis in patients with SLE is caused by conventional cardiovascular risk factors and specific ones, associated with the disease and its therapy. A slight increase in such an inflammatory marker as C-reactive protein (CRP) is thought to reflect the presence of subclinical inflammation in the vascular wall, connected with atherosclerotic process. The authors analyzed the frequency of clinical and subclinical (an increase in the thickness of intima-media complex (IMC)) atherosclerotic manifestations, the summary coronary risk, the prevalence of conventional risk factors, and CRP level in 133 female patients with SLE and in 50 healthy donors. Compared to the control group, SLE patients were younger, developed cardiovascular diseases (CAD, stenocardia, myocardial infarction, and cerebral stroke (p = 0.05) as well as arterial hypertension more often, had higher levels of hs-CRP and triglycerides, and lower levels of high density lipoprotein cholesterol (HDLC). There was a positive correlation between hs-CRP level and the activity of the disease according to ECLAM score, ES value, IgG and IgM levels, hematological disturbances (anemia, leucopenia, and/or thrombocytopenia) , and a negative correlation with total cholesterol level, HDLC there was a moderate correlation between hs-CRP and a maximal IMC value.