In international cooperation with EuroFlow PID, we will introduce uniform and optimized methodology for a detailed immunophenotyping of lymphocytes using multicolor flow cytometry (8-12 parameters) in patients with immunodeficiency, which are examinated in regional centers in Prague and Brno. The project will lead to improved diagnostics and classification of immunodeficiencies, particularly the rare types of these diseases, as well as to revealing of patophysiological mechanisms and mechanisms that modify the clinical progress of the primary and secondary immunodeficiencies. The project is designed as multicentric to cover the majority of immunodeficient patients in the Czech Republic and to reach the maximal application of the results in practice. Cooperation with Institute of Molecular Genetics of Academy of Science of the Czech Republic will allow detection of patophysiological mechanisms on detailed molecular level.

U pacientů s imunodeficiencí vyšetřeným v regionálních centrech (Praha a Brno) zavedeme v mezinárodní spolupráci s EuroFlow-PID jednotnou a optimalizovanou metodiku vyšetření podrobného imunofenotypu lymfocytů metodou mnohobarevné průtokové cytometrie (8-12 parametrů). Projekt povede ke zlepšení diagnostiky a klasifikace imunodeficiencí, především u vzácných typů onemocnění. Povede také k objevení patofysiologických mechanismů a mechanismů jež modifikují klinický průběh onemocnění (u primárních i sekundárních imunodeficiencí). Projekt je koncipován jako multicentrický, což povede jednak k pokrytí většiny imunodeficientních pacientů v ČR, jednak k maximálnímu uplatnění výsledků v praxi. Spolupráce s ÚMG AV ČR umožní odhalení patofysiologických mechanismů na detailní molekulární úrovni.

• MeSH
• algoritmy MeSH
• B-lymfocyty cytologie   MeSH
• běžná variabilní imunodeficience diagnóza   klasifikace   MeSH
• imunofenotypizace MeSH
• průtoková cytometrie MeSH
• T-lymfocyty cytologie   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• alergologie a imunologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Článek shrnuje diagnostická kritéria, etiopatogenezi, klinické projevy a léčbu primárních i sekundárních poruch tvorby protilátek. Humorální imunodeficity obecně spojuje náchylnost k infekcím způsobeným opouzdřenými baktériemi. Dominují infekce dýchacích cest a trávicího ústrojí. U řady klinických jednotek je zvýšená náchylnost k autoimunitním či alergickým onemocněním. Základem léčby jsou substituce imunoglobuliny a cílená antibiotická terapie. Díky včasnější diagnostice a lepší dostupnosti léčby se prognóza i kvalita života těchto nemocných výrazně zlepšily.

The article summarises diagnostic criteria, etiopathogenesis, clinical symptoms, and management of primary and secondary humoural immunodeficiencies. Humoural immunodeficiencies are connected with higher susceptibility to infections caused by encapsulated bacteria. Infections of respiratory and gastrointestinal tract have been predominant. There is also a trend to autoimmune complications and allergies in patients with this diagnosis. The therapy is based on immunoglobulin substitution and targeted antibiotic treatment. Due to earlier diagnosis and better availability of therapeutic options in recent years, the prognosis of patients with primary immunodeficiencies and their quality of life has improved significantly.

• Klíčová slova
• XLA, CVID, humorální, imunodeficience,
• MeSH
• agamaglobulinemie * diagnóza   etiologie   genetika   MeSH
• běžná variabilní imunodeficience genetika   klasifikace   patologie   MeSH
• diferenciální diagnóza MeSH
• dítě MeSH
• dospělí MeSH
• dysgamaglobulinemie * diagnóza   klasifikace   terapie   MeSH
• lidé MeSH
• prognóza MeSH
• protilátky MeSH
• syndromy imunologické nedostatečnosti * diagnóza   klasifikace   terapie   MeSH
• thymom komplikace   terapie   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH

The aim of this study was to find an objective computational approach for phenotype analysis of common variable immunodeficiency (CVID) patients that describes all differences in the six-color space and to form groups of patients using computational methods. CVID is a heterogeneous primary immunodeficiency disorder where molecular defect is recognized in <10% of the cases and is unknown in the majority of patients. The current CVID classification, EUROClass, is based on quantification of selected B-cell subsets. Using six-color polychromatic flow cytometry, we analyzed B-cell phenotypes in a cohort of 48 CVID patients and 49 healthy donors. We used a "probability binning" algorithm to create 1,024 bins (each bin is a six-color gate) that covered the cells' distribution within the entire B-cell compartment. A matrix file recording cellular content in all the bins was made. The hierarchical clustering of the individual samples was analyzed using a Pearson correlation of the bins' values. The Cut tree algorithm found 12 clusters. In six clusters, healthy individuals predominated; in one cluster, smB+CD21low (CVID patients by EUROClass) cells prevailed; in one cluster, smB-CD21norm cells prevailed; in one cluster, smB+CD21low cells prevailed; the remaining cluster was mixed. The overall reproducibility of probability binning clustering was confirmed by matching of replicates to the original cohort using the similarity matrix of the Pearson correlation, 15 replicates matched the same individual, three replicates matched a different individual within the same cluster, and three replicates matched to a different cluster. We were able to define B-cell subsets over- or under-represented in a particular cluster and display them back in the flow cytometry software. We describe a new analytical approach that enables a search in an objective computational environment for patient cohorts that are defined by similar B-cell profiles and thus contribute to the description of differences between CVID patient groups. Copyright 2009 International Society for Advancement of Cytometry.

• MeSH
• algoritmy MeSH
• analýza hlavních komponent MeSH
• B-lymfocyty imunologie   MeSH
• běžná variabilní imunodeficience diagnóza   klasifikace   krev   MeSH
• dospělí MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• pravděpodobnost MeSH
• průtoková cytometrie metody   MeSH
• senioři MeSH
• shluková analýza MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

The European Common Variable Immunodeficiency Disorders registry was started in 1996 to define distinct clinical phenotypes and determine overlap within individual patients. A total of 7 centers contributed patient data, resulting in the largest cohort yet reported. Patients (334), validated for the diagnosis, were followed for an average of 25.6 years (9461 patient-years). Data were used to define 5 distinct clinical phenotypes: no complications, autoimmunity, polyclonal lymphocytic infiltration, enteropathy, and lymphoid malignancy. A total of 83% of patients had only one of these phenotypes. Analysis of mortality showed a considerable reduction in the last 15 years and that different phenotypes were associated with different survival times. Types of complications and clinical phenotypes varied significantly between countries, indicating the need for large, international registries. Ages at onset of symptoms and diagnosis were shown to have a Gaussian distribution, but were not useful predictors of phenotype. The only clinical predictor was polyclonal lymphocytic infiltration, which was associated with a 5-fold increased risk of lymphoid malignancy. There was widespread variation in the levels of serum immunoglobulin isotypes as well as in the percentages and absolute numbers of B cells, confirming the heterogeneity of these conditions. Higher serum IgM and lower circulating CD8 proportions were found to be predictive markers for polyclonal lymphocytic infiltration and autoimmunity, respectively.

• MeSH
• autoimunita MeSH
• běžná variabilní imunodeficience * klasifikace   komplikace   mortalita   patologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• fenotyp MeSH
• imunoglobulinové izotypy krev   MeSH
• kohortové studie MeSH
• leukemická infiltrace MeSH
• lidé MeSH
• prognóza MeSH
• registrace MeSH
• věk při počátku nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• multicentrická studie MeSH
• práce podpořená grantem MeSH

The heterogeneity of common variable immunodeficiency (CVID) calls for a classification addressing pathogenic mechanisms as well as clinical relevance. This European multicenter trial was initiated to develop a consensus of 2 existing classification schemes based on flowcytometric B-cell phenotyping and the clinical course. The clinical evaluation of 303 patients with the established diagnosis of CVID demonstrated a significant coincidence of granulomatous disease, autoimmune cytopenia, and splenomegaly. Phenotyping of B-cell subpopulations confirmed a severe reduction of switched memory B cells in most of the patients that was associated with a higher risk for splenomegaly and granulomatous disease. An expansion of CD21(low) B cells marked patients with splenomegaly. Lymphadenopathy was significantly linked with transitional B-cell expansion. Based on these findings and pathogenic consideration of B-cell differentiation, we suggest an improved classification for CVID (EUROclass), separating patients with nearly absent B cells (less than 1%), severely reduced switched memory B cells (less than 2%), and expansion of transitional (more than 9%) or CD21(low) B cells (more than 10%). Whereas the first group contains all patients with severe defects of early B-cell differentiation, severely reduced switched memory B cells indicate a defective germinal center development as found in inducible constimulator (ICOS) or CD40L deficiency. The underlying defects of expanded transitional or CD21(low) B cells remain to be elucidated. This trial is re-gistered at http://www.uniklinik-freiburg.de/zks/live/uklregister/Oeffentlich.html as UKF000308.

• MeSH
• B-lymfocyty imunologie   patologie   MeSH
• běžná variabilní imunodeficience * epidemiologie   imunologie   klasifikace   patologie   MeSH
• dospělí MeSH
• homeostáza imunologie   MeSH
• imunofenotypizace * MeSH
• imunoglobuliny krev   MeSH
• kohortové studie MeSH
• konsensus MeSH
• lidé středního věku MeSH
• lidé MeSH
• průtoková cytometrie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

Common variable immunodeficiency (CVID) is primary hypogammaglobulinaemia with an unknown aetiopathogenesis. Although various abnormalities of T and B cells have been described, their pathogenetic roles are unclear. We determined T and B lymphocyte subsets known to be abnormal in CVID in order to disclose possible relations between numerical abnormalities in those cells. Markers associated with B cell development (CD21, CD27, IgM, IgD) were determined on B lymphocytes (CD19+); T lymphocyte development (CD45RA, CD45RO, CD62L) and activation markers (CD25, CD27, CD28, CD29, CD38, CD57, HLA-DR) were determined on CD4+ and CD8+ T lymphocytes in 42 CVID patients and in 33 healthy controls. Abnormalities in CD4+ T lymphocyte activation markers (increase in CD29, HLA-DR, CD45RO, decrease in CD27, CD62L, CD45RA) were observed particularly in patients with a decreased number of memory (CD27+) and mature (CD21+) B cells (group Ia according to the Freiburg group's classification), while abnormalities observed in CD8+ cells (increase in CD27 and CD28 and decrease in HLA-DR, CD57 and CD38) did not depend upon grouping patients together according to B lymphocyte developmental subpopulations. We observed correlations between immature B cells (IgM+ CD21-) and expression of CD27, CD62L, CD45RA, CD45RO and HLA-DR on CD4+ T cells in CVID patients but not in the control group. The expression of CD27 and CD45RA on CD4+ T lymphocytes, such as the percentage of IgD+ CD27- and IgD+ CD27+ cells in B lymphocytes, showed age dependency to be more significant than in the control group. Our study demonstrates that T and B lymphocyte abnormalities in CVID are partially related to each other. Some of those abnormalities are not definite, but may evolve with age of the patient.

• MeSH
• aktivace lymfocytů účinky léků   MeSH
• antigeny diferenciační B-lymfocytární imunologie   MeSH
• B-lymfocyty imunologie   MeSH
• běžná variabilní imunodeficience imunologie   klasifikace   MeSH
• biologické markery analýza   MeSH
• CD antigeny imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• diferenciační antigeny T-lymfocytů imunologie   MeSH
• dospělí MeSH
• financování organizované MeSH
• imunoglobulin D imunologie   MeSH
• imunoglobulin M imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• podskupiny B-lymfocytů imunologie   MeSH
• průtoková cytometrie metody   MeSH
• stárnutí imunologie   MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

• MeSH
• běžná variabilní imunodeficience klasifikace   diagnóza   terapie   MeSH
• genetické nemoci vrozené imunologie   MeSH
• nemoci imunitního systému genetika   MeSH
• syndromy imunologické nedostatečnosti klasifikace   diagnóza   terapie   MeSH
• Publikační typ
• populární práce MeSH
• příručky MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• alergologie a imunologie
• genetika, lékařská genetika

• MeSH
• B-lymfocyty imunologie   MeSH
• běžná variabilní imunodeficience imunologie   klasifikace   MeSH
• CD antigeny analýza   MeSH
• finanční podpora výzkumu jako téma MeSH
• lidé MeSH
• průtoková cytometrie metody   MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• lidé MeSH