ANCA-asociované vaskulitidy : komplexní diagnostický přístup

Cíle práce: Cílem této práce bylo zhodnotit parametry buněčné imunity u pacientů s ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies - autoprotilátky proti cytoplazmě neutrofilů)-asociovanými vaskulitidami (AAV) v různých stádiích onemocnění, s různou terapií a s ohledem na jejich dlouhodobou prognózu. Metodika: Vyšetřeno bylo 69 pacientů s AAV, 30 zdravých jedinců a 20 pacientů s chronickým onemocněním ledvin. Metodou průtokové cytometrie jsme u buněk periferní krve stanovovali následující markery: povrchové molekuly (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 a CRTH2) a intracelulární cytokiny (interferon gama (IFN), tumor nekrotizující faktor alfa (TNF), interleukin (IL)-2 a IL-4 v CD3+ T buňkách a IL-10 a IL-12 v monocytech). Výsledky: Pacienti s AAV měli ve srovnání se zdravými kontrolami snížený celkový počet lymfocytů, CD4+ buněk a CD4+CD45RA+ buněk (p<0.001). Aktivní pacienti měli zvýšenou expresi molekul CD30 a CRTH2 (p<0.05). Zvýšená exprese CCR5 přetrvávala i v remisi. Zvýšená exprese HLA-DR, expanze CD28 subpopulace a zvýšená produkce IFN byly pozorovány v remisi, ale ne u aktivní AAV. Pacienti v remisi, u kterých byl během dlouhodobého sledování zaznamenán relaps, měli významně nižší produkci IL-10 než ti bez relapsu (p<0.01). Závěry: Naše data celkově prokazují...Objectives: The aim of this study was to assess cellular immunity parameters in patients with ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies)-associated vasculitides (AAV) at different stages of the disease, with different treatment modalities, and with respect to the long-term prognosis of the patients. Methods: We examined 69 patients with AAV, 30 healthy individuals and 20 patients with chronic kidney disease. Using flow cytometry, the following markers were assessed in peripheral blood cells: surface molecules (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 and CRTH2) and intracellular cytokines (interferon gamma (IFN), tumor necrosis factor alpha (TNF), interleukin (IL)-2 and IL-4 in CD3+ T cells and IL-10 and IL-12 in monocytes). Results: Patients with AAV had decreased total number of lymphocytes, CD4+ cells, and CD4+CD45RA+ cells compared to healthy controls (p<0.001). Active patients had increased CD30 and CRTH2 expression (p<0.05). Increased CCR5 expression persisted in remission. Increased HLA-DR expression, expansion of CD28 subpopulation and increased IFN production were noted in remission but not in active disease. Patients in remission who developed a relapse during follow-up had significantly lower IL-10 production than those without relapse (p<0.01). Conclusions: Taken...Institute of Immunology and Microbiology First Faculty of Medicine Charles University in PragueÚstav Imunologie a mikrobiologie 1. LF UK a VFN v PrazeFirst Faculty of Medicine1. lékařská fakult

Impaired Deoxyribonuclease I Activity in Patients with Inflammatory Bowel Diseases

Background and Aims. Deoxyribonuclease I (DNaseI) is an endonuclease that facilitates chromatin breakdown and promotes susceptibility to autoimmune disorders. The aim of current study was to investigate serum DNase I activity in patients with inflammatory bowel diseases (IBD). Patients and Methods. A cohort of 110 IBD patients was evaluated, aged 35 ± 12 years, 77 with Crohn's disease (CD) and 33 with ulcerative colitis (UC). 50 SLE patients and 50 healthy blood donors were examined as control groups. Results. DNase I activity in IBD patients was significantly lower than in healthy individuals, but higher than in SLE patients (P < .0001). Patients with UC showed higher DNase I activity than CD patients, P = .21. DNase I activity in female patients with IBD was significantly lower than in males, P = .024; however, no differences in DNase I activity were found in relation to gender in healthy individuals. DNase I activity has shown a strong negative correlation with the serum concentration of anti-nucleosomal antibodies in the autoimmune (SLE + IBD) cohort, as well as in the separate IBD cohort. Conclusions. Reduced serum DNase I activity probably has pathogenetic consequences in IBD. Induction of autoantibodies towards nucleosomes could be a reflection of impaired DNase I activity

ANCA-Associated Vasculitides : Complex Diagnostic Approach

Objectives: The aim of this study was to assess cellular immunity parameters in patients with ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies)-associated vasculitides (AAV) at different stages of the disease, with different treatment modalities, and with respect to the long-term prognosis of the patients. Methods: We examined 69 patients with AAV, 30 healthy individuals and 20 patients with chronic kidney disease. Using flow cytometry, the following markers were assessed in peripheral blood cells: surface molecules (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 and CRTH2) and intracellular cytokines (interferon gamma (IFN), tumor necrosis factor alpha (TNF), interleukin (IL)-2 and IL-4 in CD3+ T cells and IL-10 and IL-12 in monocytes). Results: Patients with AAV had decreased total number of lymphocytes, CD4+ cells, and CD4+CD45RA+ cells compared to healthy controls (p<0.001). Active patients had increased CD30 and CRTH2 expression (p<0.05). Increased CCR5 expression persisted in remission. Increased HLA-DR expression, expansion of CD28 subpopulation and increased IFN production were noted in remission but not in active disease. Patients in remission who developed a relapse during follow-up had significantly lower IL-10 production than those without relapse (p<0.01). Conclusions: Taken..

ANCA-Associated Vasculitides : Complex Diagnostic Approach

Objectives: The aim of this study was to assess cellular immunity parameters in patients with ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies)-associated vasculitides (AAV) at different stages of the disease, with different treatment modalities, and with respect to the long-term prognosis of the patients. Methods: We examined 69 patients with AAV, 30 healthy individuals and 20 patients with chronic kidney disease. Using flow cytometry, the following markers were assessed in peripheral blood cells: surface molecules (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 and CRTH2) and intracellular cytokines (interferon gamma (IFN), tumor necrosis factor alpha (TNF), interleukin (IL)-2 and IL-4 in CD3+ T cells and IL-10 and IL-12 in monocytes). Results: Patients with AAV had decreased total number of lymphocytes, CD4+ cells, and CD4+CD45RA+ cells compared to healthy controls (p<0.001). Active patients had increased CD30 and CRTH2 expression (p<0.05). Increased CCR5 expression persisted in remission. Increased HLA-DR expression, expansion of CD28 subpopulation and increased IFN production were noted in remission but not in active disease. Patients in remission who developed a relapse during follow-up had significantly lower IL-10 production than those without relapse (p<0.01). Conclusions: Taken..

Peripheral blood lymphocytes immunophenotyping predicts disease activity in clinically isolated syndrome patients

Abstract Background Clinically isolated syndrome (CIS) represents first neurological symptoms suggestive of demyelinating lesion in the central nervous system (CNS). Currently, there are no sufficient immunological or genetic markers predicting relapse and disability progression, nor there is evidence of the efficacy of registered disease modifying treatments (DMTs), such as intramuscular interferon beta1a. The aim of the study is to evaluate immunological predictors of a relapse or disability progression. Methods One hundred and eighty one patients with CIS were treated with interferon beta1a and followed over the period of 4 years. Lymphocyte subsets were analyzed by flow cytometry. A Kaplan-Meier estimator of survival probability was used to analyze prognosis. For statistical assessment only individual differences between baseline values and values at the time of relapse or confirmed disability progression were analysed. Results Higher levels of B lymphocytes predicted relapse-free status. On the other hand, a decrease of the naïve subset of cells (CD45RA+ in CD4+) after 12, 24, and 36 months of follow-up were associated with an increased risk of confirmed disability progression. Conclusion: Our data suggest that the quantification of lymphocyte subsets in patients after the first demyelinating event suggestive of MS may be an important biomarker

Proteinase-3 and myeloperoxidase serotype in relation to demographic factors and geographic distribution in anti-neutrophil cytoplasmic antibody-associated glomerulonephritis.

BACKGROUND: In anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis, antigen specificity varies between myeloperoxidase (MPO) and proteinase 3 (PR3). This has been reported to vary in relation to age, gender, geography and extrarenal manifestations. However, studies are difficult to compare as criteria for inclusion vary. The aim of this study was to investigate the relationship between ANCA serotype, latitude, ultraviolet (UV) radiation levels, age, gender and renal function at diagnosis in a large study with uniform inclusion criteria. METHODS: Patients with biopsy-proven ANCA-associated glomerulonephritis were identified from regional or nationwide registries in 14 centres in Norway, Sweden, the UK, the Czech Republic, Croatia, Italy and the USA during the period 2000-13. UV radiation levels for 2000-13 in Europe were obtained from the Swedish Meteorological and Hydrological Institute. RESULTS: A total of 1408 patients (45.2% PR3-ANCA) were included in the study. In univariable analysis, PR3-ANCA was significantly associated with male gender {odds ratio [OR] 2.12 [95% confidence interval (CI) 1.71-2.62]}, younger age [OR per year 0.97 (95% CI 0.96-0.98)] and higher glomerular filtration rate [OR per mL/min 1.01 (95% CI 1.01-1.02); P < 0.001] at diagnosis but not with latitude or UV radiation. In multivariable logistic regression analysis, latitude and UV radiation also became significant, with higher odds for PR3-ANCA positivity at northern latitudes/lower UV radiation levels. However, the latitudinal difference in MPO:PR3 ratio is smaller than differences previously reported concerning microscopic polyangiitis and granulomatosis with polyangiitis. CONCLUSIONS: The ratio between PR3-ANCA and MPO-ANCA varies in glomerulonephritis with respect to age, gender, renal function and geographic latitude/UV radiation levels

Rituximab in adult minimal change disease and focal segmental glomerulosclerosis - What is known and what is still unknown?

Primary forms of minimal change disease and focal segmental glomerulosclerosis are rare podocytopathies and clinically characterized by nephrotic syndrome. Glucocorticoids are the cornerstone of the initial immunosuppressive treatment in these two entities. Especially among adults with minimal change disease or focal segmental glomerulosclerosis, relapses, steroid dependence or resistance are common and necessitate re-initiation of steroids and other immunosuppressants. Effective steroid-sparing therapies and introduction of less toxic immunosuppressive agents are urgently needed to reduce undesirable side effects, in particular for patients whose disease course is complex. Rituximab, a B cell depleting monoclonal antibody, is increasingly used off-label in these circumstances, despite a low level of evidence for adult patients. Hence, critical questions concerning drug-safety, long-term efficacy and the optimal regimen for rituximab-treatment remain unanswered. Evidence in the form of large, multicenter studies and randomized controlled trials are urgently needed to overcome these limitations