Sepsis is associated with a dysregulated inflammatory response to infection. Despite the activation of inflammation, an immune suppression is often observed, predisposing patients to secondary infections. Therapies directed at restoration of immunity may be considered but should be guided by the immune status of the patients. In this paper, we described the use of a high-dimensional flow cytometry (HDCyto) panel to assess the immunophenotype of patients with sepsis. We then isolated peripheral blood mononuclear cells (PBMCs) from patients with septic shock and mimicked a secondary infection by stimulating PBMCs for 4 h in vitro with lipopolysaccharide (LPS) with or without prior exposure to either IFN-γ, or LAG-3Ig. We evaluated the response by means of flow cytometry and high-resolution clustering cum differential analysis and compared the results to PBMCs from healthy donors. We observed a heterogeneous immune response in septic patients and identified two major subgroups: one characterized by hypo-responsiveness (Hypo) and another one by hyper-responsiveness (Hyper). Hypo and Hyper groups showed significant differences in the production of cytokines/chemokine and surface human leukocyte antigen-DR (HLA-DR) expression in response to LPS stimulation, which were observed across all cell types. When pre-treated with either interferon gamma (IFN-γ) or lymphocyte-activation gene 3 (LAG)-3 recombinant fusion protein (LAG-3Ig) prior to LPS stimulation, cells from the Hypo group were shown to be more responsive to both immunostimulants than cells from the Hyper group. Our results demonstrate the importance of patient stratification based on their immune status prior to any immune therapies. Once sufficiently scaled, this approach may be useful for prescribing the right immune therapy for the right patient at the right time, the key to the success of any therapy.

• MeSH
• biologické markery krev   MeSH
• CD antigeny farmakologie   MeSH
• cytokiny krev   MeSH
• fenotyp MeSH
• HLA-DR antigeny krev   MeSH
• imunofenotypizace * MeSH
• interferon gama farmakologie   MeSH
• kultivované buňky MeSH
• leukocyty mononukleární účinky léků   imunologie   metabolismus   MeSH
• lidé MeSH
• lipopolysacharidy farmakologie   MeSH
• monitorování imunologické * MeSH
• prediktivní hodnota testů MeSH
• průběh práce MeSH
• průtoková cytometrie * MeSH
• septický šok krev   diagnóza   imunologie   MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: RF and ACPA are used as diagnostic tools and their presence has been associated with clinical response to some biologic DMARDs (bDMARDs) in RA. This study compared the impact of seropositivity on drug discontinuation and effectiveness of bDMARDs in patients with RA, using head-to-head comparisons in a real-world setting. METHODS: We conducted a pooled analysis of 16 observational RA registries. Inclusion criteria were a diagnosis of RA, initiation of treatment with rituximab (RTX), abatacept (ABA), tocilizumab (TCZ) or TNF inhibitors (TNFis) and available information on RF and/or ACPA status. Drug discontinuation was analysed using Cox regression, including drug, seropositivity, their interaction, adjusting for concomitant and past treatments and patient and disease characteristics and accounting for country and calendar year of bDMARD initiation. Effectiveness was analysed using the Clinical Disease Activity Index evolution over time. RESULTS: Among the 27 583 eligible patients, the association of seropositivity with drug discontinuation differed across bDMARDs (P for interaction <0.001). The adjusted hazard ratios for seropositive compared with seronegative patients were 1.01 (95% CI 0.95, 1.07) for TNFis, 0.89 (0.78, 1.02)] for TCZ, 0.80 (0.72, 0.88) for ABA and 0.70 (0.59, 0.84) for RTX. Adjusted differences in remission and low disease activity rates between seropositive and seronegative patients followed the same pattern, with no difference in TNFis, a small difference in TCZ, a larger difference in ABA and the largest difference in RTX (Lundex remission difference +5.9%, low disease activity difference +11.6%). CONCLUSION: Seropositivity was associated with increased effectiveness of non-TNFi bDMARDs, especially RTX and ABA, but not TNFis.

• MeSH
• antirevmatika * klasifikace   imunologie   terapeutické užití   MeSH
• biologické přípravky * klasifikace   imunologie   terapeutické užití   MeSH
• lékové interakce imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezinárodní spolupráce MeSH
• monitorování imunologické * metody   statistika a číselné údaje   MeSH
• nenasazení léčby statistika a číselné údaje   MeSH
• posouzení stavu pacienta MeSH
• registrace statistika a číselné údaje   MeSH
• revmatoidní artritida * diagnóza   farmakoterapie   epidemiologie   imunologie   MeSH
• revmatoidní faktor krev   MeSH
• trvání terapie MeSH
• výběr pacientů MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

• MeSH
• alergeny aplikace a dávkování   imunologie   MeSH
• biologické markery analýza   MeSH
• bronchiální astma * diagnóza   imunologie   terapie   MeSH
• desenzibilizace imunologická * metody   normy   MeSH
• dítě MeSH
• klinické zkoušky jako téma MeSH
• lidé MeSH
• monitorování imunologické metody   normy   MeSH
• prognóza MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• přehledy MeSH

Living donor kidney transplantation represents the best therapy of end stage renal disease. Recently, the acute rejection incidence has been observed to be higher than in deceased donor transplantation, mainly because of increasing number of genetically unrelated procedures. Prediction of acute rejection with recently available techniques often fails. The aim of this project is to analyze recipient alloreactivty using multilevel laboratory platforms to better predict risks of acute rejection, both T cell- and antibody mediated. In this project pre and posttransplant analyzes will be performed to detect anti HLA antibodies, T and B cell donor specific alloreactivity, specific populations of T and B cells along with their peripheral transcripts. Data will be analyzed in relation to postransplant outcome and distinct rejection phenotype.

Transplantace ledviny od žijících dárců představují nejlepší metodu léčby nezvratného selhání ledvin. Výskyt akutních rejekcí je v tomto programu nyní vyšší než u kadaverosních transplantací, především v důsledku rozšíření geneticky nepříbuzných transplantací. Riziko vzniku rejekce se pomocí běžně používaných metod (stanovení panel reaktivních protilátek a crossmatch) dá odhadnout jen velmi těžko. Cílem této studie je posoudit aloreaktivitu příjemce pomocí panelu laboratorních testů a odhadnout riziko vzniku T buňkami a protilátkami zprostředkované rejekce. Předmětem výzkumu je detekce předtransplantačních a potransplantačních anti HLA protilátek spolu s určením dárcovsky specifické T a B lymfocytární aloreaktivity (ELISPOT), vyšetření periferních populací T a B lymfocytů a jejich specifických periferních transkriptů. Získaná data budou analyzována ve vztahu k potransplantačnímu průběhu a fenotypu rejekce.

• MeSH
• B-lymfocyty MeSH
• ELISPOT MeSH
• monitorování imunologické MeSH
• protilátky škodlivé účinky   MeSH
• rejekce štěpu diagnóza   MeSH
• T-lymfocyty MeSH
• transplantace ledvin škodlivé účinky   MeSH
• žijící dárci MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• transplantologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

BACKGROUND & AIMS: γδ T cells comprise a substantial proportion of tissue-associated lymphocytes. However, our current understanding of human γδ T cells is primarily based on peripheral blood subsets, while the immunobiology of tissue-associated subsets remains largely unclear. Therefore, we aimed to elucidate the T cell receptor (TCR) diversity, immunophenotype and function of γδ T cells in the human liver. METHODS: We characterised the TCR repertoire, immunophenotype and function of human liver infiltrating γδ T cells, by TCR sequencing analysis, flow cytometry, in situ hybridisation and immunohistochemistry. We focussed on the predominant tissue-associated Vδ2- γδ subset, which is implicated in liver immunopathology. RESULTS: Intrahepatic Vδ2- γδ T cells were highly clonally focussed, with single expanded clonotypes featuring complex, private TCR rearrangements frequently dominating the compartment. Such T cells were predominantly CD27lo/- effector lymphocytes, whereas naïve CD27hi, TCR-diverse populations present in matched blood were generally absent in the liver. Furthermore, while a CD45RAhi Vδ2- γδ effector subset present in both liver and peripheral blood contained overlapping TCR clonotypes, the liver Vδ2- γδ T cell pool also included a phenotypically distinct CD45RAlo effector compartment that was enriched for expression of the tissue tropism marker CD69, the hepatic homing chemokine receptors CXCR3 and CXCR6, and liver-restricted TCR clonotypes, suggestive of intrahepatic tissue residency. Liver infiltrating Vδ2- γδ cells were capable of polyfunctional cytokine secretion, and unlike peripheral blood subsets, were responsive to both TCR and innate stimuli. CONCLUSION: These findings suggest that the ability of Vδ2- γδ T cells to undergo clonotypic expansion and differentiation is crucial in permitting access to solid tissues, such as the liver, which results in functionally distinct peripheral and liver-resident memory γδ T cell subsets. They also highlight the inherent functional plasticity within the Vδ2- γδ T cell compartment and provide information that could be used for the design of cellular therapies that suppress liver inflammation or combat liver cancer. LAY SUMMARY: γδ T cells are frequently enriched in many solid tissues, however the immunobiology of such tissue-associated subsets in humans has remained unclear. We show that intrahepatic γδ T cells are enriched for clonally expanded effector T cells, whereas naïve γδ T cells are largely excluded. Moreover, whereas a distinct proportion of circulating T cell clonotypes was present in both the liver tissue and peripheral blood, a functionally and clonotypically distinct population of liver-resident γδ T cells was also evident. Our findings suggest that factors triggering γδ T cell clonal selection and differentiation, such as infection, can drive enrichment of γδ T cells into liver tissue, allowing the development of functionally distinct tissue-restricted memory populations specialised in local hepatic immunosurveillance.

• MeSH
• buněčná diferenciace imunologie   MeSH
• imunologická paměť fyziologie   MeSH
• intraepiteliální lymfocyty * imunologie   patologie   MeSH
• játra * imunologie   patologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• monitorování imunologické metody   MeSH
• receptory antigenů T-buněk gama-delta imunologie   MeSH
• T-lymfocyty - podskupiny imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cytomegalovirus (CMV) infection influences both short and long term outcomes in immunosuppressed organ transplant recipients. The aim of this study was to evaluate the effect of different induction immunosuppression regimens on CMV specific T cell response in patients with already established CMV immunity. In 24 seropositive living donor kidney recipients, the frequency of CMV specific T cells was determined by ELISPOT (Enzyme-Linked ImmunoSpot) assay prior and 6 months after transplantation. Recipients' peripheral blood mononuclear cells were stimulated with immediate-early (IE1) and phosphoprotein 65 (pp65) CMV-derived peptide pools and the number of cells producing interferon gamma (IFN-gamma) was assessed. Patients received quadruple immunosuppression based either on depletive rabbit antithymocyte globulin (rATG) or non-depletive basiliximab induction and tacrolimus/mycophenolate mofetil/steroids. Patients with rATG induction received valgancyclovir prophylaxis. No effects of different induction agents on CMV specific T cell immunity were found at sixth month after kidney transplantation. There were no associations among dialysis vintage, pretransplant CMV specific T cell immunity, and later CMV DNAemia. Similarly, no effect of CMV prophylaxis on CMV specific T cell immunity was revealed. This study shows no effect of posttransplant immunosuppression on CMV specific T cell immunity in living donor kidney transplant recipients with CMV immunity already established, regardless of lymphocyte depletion and CMV prophylaxis.

• MeSH
• buněčná imunita MeSH
• cytomegalovirové infekce prevence a kontrola   MeSH
• Cytomegalovirus imunologie   MeSH
• dospělí MeSH
• fosfoproteiny imunologie   MeSH
• imunosupresiva terapeutické užití   MeSH
• imunosupresivní léčba metody   MeSH
• indukční chemoterapie MeSH
• lidé středního věku MeSH
• lidé MeSH
• monitorování imunologické MeSH
• proteiny virové matrix imunologie   MeSH
• T-lymfocyty imunologie   MeSH
• thymocyty imunologie   MeSH
• transplantace ledvin metody   MeSH
• žijící dárci * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• alergie * imunologie   terapie   MeSH
• hodnocení rizik MeSH
• imunitní systém MeSH
• imunoterapie * metody   MeSH
• lidé MeSH
• monitorování imunologické MeSH
• monoklonální protilátky * škodlivé účinky   terapeutické užití   MeSH
• nádory * imunologie   terapie   MeSH
• nemoci imunitního systému * imunologie   terapie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: There is high medical need for safe long-term immunosuppression monotherapy in kidney transplantation. Selective targeting of post-transplant alloantigen-(re)activated effector-T cells by anti-TNF antibodies after global T cell depletion may allow safe drug minimization, however, it is unsolved what might be the best maintenance monotherapy. METHODS: In this open, prospective observational single-centre trial, 20 primary deceased donor kidney transplant recipients received 2x20 mg Alemtuzumab (d0/d1) followed by 5 mg/kg Infliximab (d2). For 14 days all patients received only tacrolimus, then they were allocated to either receive tacrolimus (TAC, n = 13) or sirolimus (SIR, n = 7) monotherapy, respectively. Protocol biopsies and extensive immune monitoring were performed and patients were followed-up for 60 months. RESULTS: TAC-monotherapy resulted in excellent graft survival (5yr 92%, 95%CI: 56.6-98.9) and function, normal histology, and no proteinuria. Immune monitoring revealed low intragraft inflammation (urinary IP-10) and hints for the development of operational tolerance signature in the TAC- but not SIR-group. Remarkably, the TAC-monotherapy was successful in all five presensitized (ELISPOT+) patients. However, recruitment into SIR-arm was stopped (after n = 7) because of high incidence of proteinuria and acute/chronic rejection in biopsies. No opportunistic infections occurred during follow-up. CONCLUSIONS: In conclusion, our novel fast-track TAC-monotherapy protocol is likely to be safe and preliminary results indicated an excellent 5-year outcome, however, a full-scale study will be needed to confirm our findings. TRIAL REGISTRATION: EudraCT Number: 2006-003110-18.

• MeSH
• antigeny nádorové MeSH
• biologické markery metabolismus   MeSH
• CD antigeny MeSH
• dospělí MeSH
• glykoproteiny antagonisté a inhibitory   MeSH
• imunosupresiva terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• monitorování imunologické MeSH
• prospektivní studie MeSH
• rejekce štěpu farmakoterapie   etiologie   MeSH
• sirolimus terapeutické užití   MeSH
• stanovení celkové genové exprese MeSH
• takrolimus terapeutické užití   MeSH
• TNF-alfa antagonisté a inhibitory   MeSH
• transplantace ledvin škodlivé účinky   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• pozorovací studie MeSH

Project is focused on the analysis of the HLA antibody profile in patients following kidney transplantation with the use of more sensitive and modern solid-phase technology on micro-beads (Luminex technique). 1) Groups of recipients with/without antibody-mediated allograft rejection will be compared with the aim to evaluate the clinical relevance of particular HLA antibody types for prediction of the allograft rejection risk. Pre-transplant HLA antibody analysis might be useful for stratification of recipients into the groups with low/high rejection risk and with the use of different strategy of immunosuppressive therapy or post-transplant monitoring. 2) Detailed post-transplant monitoring of HLA antibody production might be useful for earlier detection of the allograft rejection development. Followed by the individualized intensive anti-rejection therapy, this approach might save the transplanted kidney. The project will propose the optimal schedule for allograft rejection post-transplant monitoring.

Projekt je zaměřen na analýzu profilu HLA protilátek u pacientů před a po transplantaci ledviny s použitím senzitivnější techniky jejich stanovení (Luminex). 1) Srovnáním skupin pacientů s/bez protilátkami zprostředkované rejekce štěpu bude hodnocen klinický význam jednotlivých typů HLA protilátek detekovaných nově zavedenou senzitivnější technikou pro předpověď stupně rizika rozvoje rejekce štěpu. Vyšetření HLA protilátek před transplantací poskytne možnost stratifikace pacientů do skupin s nízkým/vysokým rizikem rejekce štěpu a s možností různé strategie imunosupresivní terapie nebo potrasplantačního monitorování. 2) Podrobným monitorováním dynamiky tvorby HLA protilátek citlivějšími technikami v potransplantačním období je snaha dosáhnout včasného záchytu rozvíjející se rejekce štěpu s možností individualizace antirejekční terapie a záchrany transplantované ledviny. Projekt umožní nastavení optimálního časového harmonogramu monitorování rejekce štěpu v transplantačním protokolu.

• MeSH
• cytotoxicita imunologická MeSH
• HLA antigeny analýza   MeSH
• monitorování imunologické MeSH
• průtoková cytometrie MeSH
• rejekce štěpu MeSH
• specificita protilátek MeSH
• transplantační tolerance MeSH

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

The capability of lymphocytes to respond to antigenic or mitogenic stimulation is an important feature in the diagnosis of various immunodeficiencies and immune disorders. We used large cohorts of both immune compromised patients and healthy controls to measure lymphocyte proliferations by means of three methods: CFSE staining, Ki-67 expression and (3)H-thymidine incorporation. The advantages and disadvantages of each method was then evaluated for use in routine clinical diagnostic. The statistical analysis was performed between the outcomes and the correlation between all three methods was computed. CFSE and Ki-67 assay correlated well with the r=0.767, correlation between Ki-67 expression and (3)H-thymidine incorporation was 0.546 and correlation between CFSE staining and (3)H-thymidine incorporation was 0.337. The differences between these three methods concerning complexity, sensitivity and reliability as well as the financial aspects are discussed hereafter. CFSE and its analogues provide the cheapest and reasonable choice for measuring lymphocyte proliferation, while Ki-67 represents a more expensive, but more sensitive and robust method. The original (3)H-thymidine assay does not bring any advantages and cannot compare to the competition presented by modern flow cytometric methods available today.

• MeSH
• aktivace lymfocytů MeSH
• antigen Ki-67 metabolismus   MeSH
• fluoresceiny MeSH
• hostitel s imunodeficiencí * MeSH
• imunoanalýza metody   MeSH
• kohortové studie MeSH
• kultivované buňky MeSH
• lidé MeSH
• lymfocyty imunologie   MeSH
• monitorování imunologické MeSH
• proliferace buněk MeSH
• průtoková cytometrie MeSH
• reprodukovatelnost výsledků MeSH
• senzitivita a specificita MeSH
• sukcinimidy MeSH
• thymidin MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH