The EuroFlow PID consortium developed a set of flow cytometry tests for evaluation of patients with suspicion of primary immunodeficiency (PID). In this technical report we evaluate the performance of the SCID-RTE tube that explores the presence of recent thymic emigrants (RTE) together with T-cell activation status and maturation stages and discuss its applicability in the context of the broader EuroFlow PID flow cytometry testing algorithm for diagnostic orientation of PID of the lymphoid system. We have analyzed peripheral blood cells of 26 patients diagnosed between birth and 2 years of age with a genetically defined primary immunodeficiency disorder: 15 severe combined immunodeficiency (SCID) patients had disease-causing mutations in RAG1 or RAG2 (n = 4, two of them presented with Omenn syndrome), IL2RG (n = 4, one of them with confirmed maternal engraftment), NHEJ1 (n = 1), CD3E (n = 1), ADA (n = 1), JAK3 (n = 3, two of them with maternal engraftment) and DCLRE1C (n = 1) and 11 other PID patients had diverse molecular defects [ZAP70 (n = 1), WAS (n = 2), PNP (n = 1), FOXP3 (n = 1), del22q11.2 (DiGeorge n = 4), CDC42 (n = 1) and FAS (n = 1)]. In addition, 44 healthy controls in the same age group were analyzed using the SCID-RTE tube in four EuroFlow laboratories using a standardized 8-color approach. RTE were defined as CD62L+CD45RO-HLA-DR-CD31+ and the activation status was assessed by the expression of HLA-DR+. Naïve CD8+ T-lymphocytes and naïve CD4+ T-lymphocytes were defined as CD62L+CD45RO-HLA-DR-. With the SCID-RTE tube, we identified patients with PID by low levels or absence of RTE in comparison to controls as well as low levels of naïve CD4+ and naïve CD8+ lymphocytes. These parameters yielded 100% sensitivity for SCID. All SCID patients had absence of RTE, including the patients with confirmed maternal engraftment or oligoclonally expanded T-cells characteristic for Omenn syndrome. Another dominant finding was the increased numbers of activated CD4+HLA-DR+ and CD8+HLA-DR+ lymphocytes. Therefore, the EuroFlow SCID-RTE tube together with the previously published PIDOT tube form a sensitive and complete cytometric diagnostic test suitable for patients suspected of severe PID (SCID or CID) as well as for children identified via newborn screening programs for SCID with low or absent T-cell receptor excision circles (TRECs).

• Klíčová slova
• EuroFlow, diagnosis, flow cytometric immunophenotyping, primary immunodeficiencies (PID), severe combined immune deficiency (SCID), standardization,
• MeSH
• HLA-DR antigeny analýza   MeSH
• imunofenotypizace metody   MeSH
• kojenec MeSH
• lidé MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• primární imunodeficience diagnóza   imunologie   MeSH
• průtoková cytometrie metody   MeSH
• T-lymfocyty imunologie   MeSH
• těžká kombinovaná imunodeficience imunologie   MeSH
• thymus imunologie   MeSH
• Check Tag
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• HLA-DR antigeny MeSH

The aim was to analyze T-regulatory cells (Tregs), activated CD8(+) T cells, and transforming growth factor-beta (TGF)-β in hepatitis C patients. We enrolled 31 patients with chronic genotype 1 hepatitis C virus (HCV) infection, 30 seropositive persons with spontaneous HCV elimination, and 23 healthy volunteers. The patients were examined at the beginning of the interferon-alpha (IFN-α)-based therapy (baseline) and at weeks 4 (W4) and 12 (W12) of the therapy. The percentage of Tregs and the expression of activation markers CD38 and HLA-DR on CD8(+) T cells were analyzed in the peripheral blood by flow cytometry. Serum levels of TGF-β were measured in a multiplex assay using flow cytometry. The percentage of Tregs in patients was higher than in controls and seropositive persons. Similarly, the percentage of CD8(+) T cells expressing CD38 and HLA-DR was higher in patients compared with controls and seropositive persons. Chronic HCV infection is associated with elevated circulating Tregs and activated CD8(+) T cells. During IFN-α-based therapy these cells gradually increase, whereas TGF-β serum levels decrease.

• Klíčová slova
• CD38, Hepatitis C virus, interferon, regulatory T cells, transforming growth factor-beta,
• MeSH
• aktivace lymfocytů * MeSH
• antigeny CD38 analýza   MeSH
• antivirové látky terapeutické užití   MeSH
• CD8-pozitivní T-lymfocyty chemie   imunologie   MeSH
• chronická hepatitida C farmakoterapie   patologie   virologie   MeSH
• dospělí MeSH
• genotyp MeSH
• Hepacivirus klasifikace   genetika   MeSH
• HLA-DR antigeny analýza   MeSH
• imunofenotypizace MeSH
• inhibitory proteas terapeutické užití   MeSH
• interferon alfa terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny analýza   MeSH
• mladý dospělý MeSH
• průtoková cytometrie MeSH
• regulační T-lymfocyty imunologie   MeSH
• ribavirin terapeutické užití   MeSH
• senioři MeSH
• sérum chemie   MeSH
• transformující růstový faktor beta krev   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny CD38 MeSH
• antivirové látky MeSH
• CD38 protein, human MeSH Prohlížeč
• HLA-DR antigeny MeSH
• inhibitory proteas MeSH
• interferon alfa MeSH
• membránové glykoproteiny MeSH
• ribavirin MeSH
• transformující růstový faktor beta MeSH

OBJECTIVE: We measured and compared changes in the percentage of cells expressing CD80, CD86, CD40, HLA-DR and the expression of these molecules on B cells and monocytes of patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery. METHODS: Blood samples from patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery were collected before surgery, instantly after surgery and on the 1(st), 3(rd) and 7(th) days after surgery. Surface expression of CD80, CD86, CD40 and HLA-DR molecules was determined by flow cytometry. RESULTS: Our results show that all three surgical techniques altered the expression of these molecules, as well as the percentage relative number of specific cell populations. We identified statistically significant differences when comparing different surgical techniques. On-pump surgery revealed a more pronounced impact on the phenotype of immune system cells than the other techniques. Therefore, it is likely that the function of immune cells is changed the most by on-pump surgery. We found a lower decrease in the number of CD80(+) monocytes and a lower drop in the CD40 expression on monocytes in off-pump patients in comparison with on-pump patients. CONCLUSION: All the types of cardiac surgical techniques, off-pump, on-pump and modified mini-invasive on-pump, are associated with changes in CD80, CD86, CD40 and HLA-DR expression. We found several significant differences in the expression of the selected molecules when we compared all three groups of patients.

• Klíčová slova
• CD40, CD80, CD86, HLA-DR, cardiac surgery, mini-invasive, off-pump, on-pump,
• MeSH
• antigeny CD40 analýza   MeSH
• antigeny CD80 analýza   MeSH
• antigeny CD86 analýza   MeSH
• B-lymfocyty imunologie   MeSH
• HLA-DR antigeny analýza   MeSH
• kardiochirurgické výkony * MeSH
• lidé středního věku MeSH
• lidé MeSH
• monocyty imunologie   MeSH
• prospektivní studie MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny CD40 MeSH
• antigeny CD80 MeSH
• antigeny CD86 MeSH
• HLA-DR antigeny MeSH

BACKGROUND: HLA-A,B,C and HLA-D molecules present antigenic peptides to the antigen-specific receptor of autologous T lymphocytes. T-cell-mediated host-versus-tumor response might therefore depend on the presence of these molecules on tumor cells, although the absence of HLA-A,B,C determinants on a cell has been shown to increase its susceptibility to lysis by natural killer cells. The prognostic role of tumor stage and grade is well- established in colorectal cancer. In this study we used immunohistochemistry to analyse the expression of HLA-DR on epithelial cells of normal colonic mucosa, tubulovillous adenoma, and invasive carcinoma, as well as the magnitude of the stromal T lymphocytes at the relevant sites. HLA-DR expression was correlated to histological grade and Dukes stage in the cases of invasive cancer. Yet, we investigated the association of HLA-DR plus DQ genes and adenoma or carcinoma by PCR. MATERIALS AND METHODS: 31 cases of normal colonic mucosa, 12 cases of tubulovillous adenoma, and 39 cases of invasive carcinoma were surveyed for the detection of HLA-DR monoclonal antigen, and the T helper (TH) marker (CD4) in the stroma (lamina propria) of the relevant cases. RESULTS: HLA-DR was expressed in 20 of 31 normal colonic mucosas (64.5%), 4 of 12 adenomas (33.3%), and in 10 of 39 invasive carcinomas (25.6%). A strong relation of HLA-DR expression and histological grade was found (p < 0.001), but no association with Dukes stage (p = 0.141). No significant correlation between HLA-DR plus DQ genes and adenoma or cancer of the colon was found. CD4 positive cells were found in 9 of 31 normal colonic mucosas (29%), 5 of 12 adenomas (42%), and in 26 of 39 invasive carcinomas (67%). CONCLUSIONS: The results showed an inverse correlation between the expression of HLA-DR and the number of CD4 positive cells as the lesion progressed to malignancy. HLA-DR was significantly associated with tumor grade but not with Dukes stage in colonic cancer hosts. HLA-DR and DQ genes do not contribute to a susceptibility to adenoma or carcinoma.

• MeSH
• CD4-pozitivní T-lymfocyty patologie   MeSH
• HLA-DR antigeny analýza   MeSH
• imunohistochemie MeSH
• karcinom imunologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory tračníku imunologie   patologie   MeSH
• senioři MeSH
• střevní sliznice imunologie   patologie   MeSH
• vilózní adenom imunologie   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• duplikátní publikace MeSH
• Názvy látek
• HLA-DR antigeny MeSH

BACKGROUND: Sepsis is a serious disease with a high case fatality rate. A variety of changes in the host immune responsiveness are observed in the pathogenesis of sepsis, ranging from detrimental hyperinflammation to profound immunoparalysis, i.e. acquired immunodeficiency. The level of monocyte HLA-DR expression reflects the functional status of monocytes as antigen-presenting cells and granulocyte CD64 expression is also indicative of infectious inflammation. MATERIAL AND METHODS: Monocyte HLA-DR expression and granulocyte CD64 expression were measured in 49 septic patients and 30 healthy controls using flow cytometry focused on three parameters: positive cell percentage, mean fluorescence intensity and quantitation of antibodies bound per cell (QuantiBRITE). RESULTS: The significance of both monocyte HLA-DR expression and granulocyte CD64 expression in septic patients was confirmed. Monocyte HLA-DR dramatically decreases in septic patients compared to controls, is one of the prognostic factors and correlates with C-reactive protein. In contrast, granulocyte CD64 sharply rises in patients with sepsis and correlates with mediators of systemic inflammation (procalcitonin - PCT), proinflammatory mediators (interleukin-6 - IL-6, lipopolysaccharide binding protein - LBP) and anti-inflammatory cytokines (interleukin-10 - IL10). CONCLUSION: Quantitative monocyte HLA-DR expression and granulocyte CD64 expression are useful indicators in septic patients when considered along with the panel of other markers, monitored over a period of time and in the context of the clinical course of sepsis.

• MeSH
• dospělí MeSH
• granulocyty imunologie   MeSH
• HLA-DR antigeny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• monocyty imunologie   MeSH
• průtoková cytometrie * MeSH
• receptory IgG analýza   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• sepse imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• HLA-DR antigeny MeSH
• receptory IgG MeSH

AIM: Breast cancer is a frequent cause of death among women with gynaecologic malignancies despite the introduction of combination chemotherapy. There is therefore a need for new therapeutic strategies for patients with breast cancer, such as cellular immunotherapy. In this immunohistochemical study we analyzed the epithelial expression of major histocompatibility complex (MHC) class II (HLA-DR) on atypical and malignant primary mammary epithelial cells, as well as the magnitude of the stromal T lymphocytes (T4 subset) at the tumor site. EXPERIMENTAL DESIGN: The study was carried out retrospectively in tumor tissue from 82 patients with mammary lesions (31 cases of atypical ductal hyperplasia -ADH-, 12 cases of ductal carcinoma in situ -DCIS- and 39 cases of infiltrating ductal carcinoma not otherwise specified -IDC-NOS). Medullary carcinomas were not included in our investigation. Material used had been formalin fixed and paraffin embedded. RESULTS: HLA class II (DR) was expressed in 20 of 31 ADHs (64.5%), in 4 of 12 DCISs (33.3%), and in 10 of 39 IDC-NOSs (25.6%). CD4 was expressed in 9 of 31 ADHs (29%), in 5 of 12 DCISs (42%), and in 26 of 39 IDC-NOSs (67%). CONCLUSIONS: The results showed decreased epithelial expression of HLA class II (DR) and increased stromal expression of CD4, as the lesion progressed to malignancy. Gradual loss of epithelial HLA class II expression might be a manifestation of cellular differentiation from the atypical form versus the malignant one, signaling simultaneously a selective effect on the response capacity of the immune system.

• MeSH
• antigeny CD4 analýza   MeSH
• dospělí MeSH
• duktální karcinom prsu imunologie   MeSH
• HLA-DR antigeny analýza   MeSH
• hyperplazie imunologie   MeSH
• intraduktální neinfiltrující karcinom imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu imunologie   MeSH
• počet CD4 lymfocytů * MeSH
• prsy patologie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny CD4 MeSH
• HLA-DR antigeny MeSH

UNLABELLED: Fifty-seven children (f/m=31/26) who survived diarrhea (D) + hemolytic uremic syndrome (HUS) were evaluated. The examinations were performed 1-27 years (median 7 years) from the onset of the acute disease. Patients aged 2.3-27 years (median 10 years) were allocated to three groups: Recovery (R, complete recovery), Residual renal symptoms (RRS, hematuria and/or proteinuria and/or hypertension with glomerular filtration rate (GFR) >80 ml/min/1.73 m(2), or moderate renal insufficiency with slightly decreased GFR to 60-80 ml/min/1.73 m(2) with or without residual renal symptoms), and Chronic renal insufficiency/failure (CRI/F, dialysis, transplantation - GFR <60 ml/min/ 1.73 m(2)). Results from 18 patients who survived more than 10 years after HUS demonstrated a high prevalence of renal damage. Only 6/18 patients were in group R, 7/18 patients were in group RRS and 5/18 patients were in group CRI/F. An early onset of HUS (36 patients between 0 and 2 years) was associated with a better prognosis when compared with late onset (21 patients aged more than 2 years), P=0.009. Serology typing of Human leukocyte antigens (HLA) classes I and II in 64 patients revealed a significantly higher frequency of DR9 antigen ( P=0.0037) and a lower frequency of DQ1 antigen ( P=0.009) in D+HUS patients compared with healthy Czech blood donors. CONCLUSION: Our study demonstrates a high prevalence of late renal damage in Czech patients surviving after D+HUS. The HLA typing in our group revealed a significantly higher rate of HLA DR9 haplotypes in D+HUS patients.

• MeSH
• chronické selhání ledvin epidemiologie   etiologie   MeSH
• dítě MeSH
• dospělí MeSH
• hemolyticko-uremický syndrom komplikace   epidemiologie   MeSH
• HLA-DQ antigeny analýza   MeSH
• HLA-DR antigeny analýza   MeSH
• HLA-DR sérologické podskupiny MeSH
• lidé MeSH
• mladiství MeSH
• následné studie MeSH
• nemoci ledvin epidemiologie   etiologie   MeSH
• předškolní dítě MeSH
• prevalence MeSH
• prognóza MeSH
• průjem etiologie   MeSH
• referenční hodnoty MeSH
• věk při počátku nemoci MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• HLA-DQ antigeny MeSH
• HLA-DQ1 antigen MeSH Prohlížeč
• HLA-DR antigeny MeSH
• HLA-DR sérologické podskupiny MeSH
• HLA-DR9 antigen MeSH Prohlížeč

OBJECTIVES: FNAB is a cytological procedure enabling the monitoring of inflammatory cells in the graft, morphological modification of parenchymal cells, and expression of antigens on aspirated cells. The aim of the study was to evaluate whether Daclizumab influences the composition of inflammatory infiltrate and expression of HLA-DR antigens and intercellular adhesive molecule ICAM-1 on parenchymal cells. METHODS: Two groups of liver allograft recipients were included and they were treated with two different immunosuppressive protocols. The first group with quadruple immunosuppression therapy (Cyclosporine A, Mycophenolate mofetil, steroids, and Daclizumab). The second group with quadruple combination immunosuppression (Cyclosporine A, Azathioprine, steroids and ATG-Fresenius). FNAB and blood samples were collected simultaneously. Corrected increments of inflammatory cells were statistically evaluated as well as expression of HLA-DR antigens and ICAM-1 on parenchymal cells. RESULTS: FNAB specimens from the Daclizumab group demonstrated significantly lower values of the total corrected increment, the corrected increment of monocytes, number of blast cells per slide, and a lower number of ICAM-1 expressing parenchymal cells. CONCLUSION: We summarise that Daclizumab significantly reduces inflammatory cells in liver graft, as well as expression of ICAM-1 on parenchymal cells.

• MeSH
• daklizumab MeSH
• dospělí MeSH
• HLA-DR antigeny analýza   MeSH
• humanizované monoklonální protilátky MeSH
• imunoglobulin G terapeutické užití   MeSH
• imunosupresiva terapeutické užití   MeSH
• jehlová biopsie MeSH
• lidé MeSH
• monoklonální protilátky terapeutické užití   MeSH
• počet leukocytů MeSH
• retrospektivní studie MeSH
• transplantace jater imunologie   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• daklizumab MeSH
• HLA-DR antigeny MeSH
• humanizované monoklonální protilátky MeSH
• imunoglobulin G MeSH
• imunosupresiva MeSH
• monoklonální protilátky MeSH

Interactions of macrophages with epithelium represent one of the pathways involved in regulating local immune mechanisms. We studied the effect of cell-cell contact with an epithelial monolayer on the phenotype of macrophages. Human monocytes and THP-1 macrophages were co-cultured with monolayers of human bronchial epithelial cells (HBECs), the alveolar type II-like cell line A549, renal adenocarcinoma epithelial cells (RA), and the lung fibroblast strain HFL-1. The expression of CD11b, CD14, CD54, and HLA-DR was measured by immunocytochemistry and flow cytometry and showed epithelial cell induction of CD54 and HLA-DR in monocytes and of all antigens in THP-1 cells. Co-culture with fibroblasts did not change the phenotype of macrophages. Separation by a filter insert inhibited most of the effects. Culture supernatants did not induce prominent phenotypic changes. Cell-cell contacts with epithelium appear to be of importance in regulating the phenotype of macrophages.

• MeSH
• antigeny CD14 analýza   MeSH
• epitelové buňky imunologie   MeSH
• HLA-DR antigeny analýza   MeSH
• imunofenotypizace MeSH
• kokultivační techniky MeSH
• ledviny cytologie   MeSH
• lidé MeSH
• makrofágový antigen 1 analýza   MeSH
• makrofágy imunologie   MeSH
• mezibuněčná adhezivní molekula-1 analýza   MeSH
• mezibuněčná komunikace imunologie   MeSH
• monocyty imunologie   MeSH
• nádorové buňky kultivované MeSH
• plíce cytologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Názvy látek
• antigeny CD14 MeSH
• HLA-DR antigeny MeSH
• makrofágový antigen 1 MeSH
• mezibuněčná adhezivní molekula-1 MeSH

BACKGROUND: To assess the prevalence of markers of autoimmune insulitis (AII) in patients classified originally as having Type-2 diabetes mellitus (Type-2 DM). 386 patients subdivided according to the BMI, C-peptide and type of treatment. METHODS AND RESULTS: Age, BMI, C-peptide, Glutamic acid decarboxylase autoantibodies (GADA), HLA-DR/,-DQ alleles. Prevalence of GADA varied from < 5% in obese patients with normal/increased C-peptide to > 30% in non-obese patients with low C-peptide. In majority of GADA positive patients, the Type-1 DM high-risk HLA-DRB1*, HLA-DQB1* alleles have been found. Among them HLA-DRB1*0302 and HLA-DRB1*0201 were more frequent than HLA-DRB1*040x and HL:A-DQB1*0302. CONCLUSIONS: Significant fraction of patients classified initially as Type-2 DM may have in fact Type-1 DM. Such patients can be recognized on the basis of assessment of serological (GADA) and immuno-genetical (HLA-DR/,-DQ alleles) markers. In some patients clinical, metabolic, immune, and immunogenetic markers may disagree. This divergence stresses multifactorial genesis of diabetes. Moreover, it can also suggest that both autoimmune insulitis and insulin resistance may coexist in parallel.

• MeSH
• autoimunitní nemoci diagnóza   MeSH
• autoprotilátky analýza   MeSH
• biologické markery analýza   MeSH
• C-peptid krev   MeSH
• diabetes mellitus 1. typu diagnóza   MeSH
• diabetes mellitus 2. typu diagnóza   MeSH
• dospělí MeSH
• glutamát dekarboxyláza imunologie   MeSH
• HLA-DQ antigeny analýza   MeSH
• HLA-DR antigeny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• autoprotilátky MeSH
• biologické markery MeSH
• C-peptid MeSH
• glutamát dekarboxyláza MeSH
• HLA-DQ antigeny MeSH
• HLA-DR antigeny MeSH