The accumulation of protein aggregates is toxic and linked to different diseases such as neurodegenerative disorders, but the role of the immune system to target and destroy aggregate-carrying cells is still relatively unknown. Here we show a substrate-specific presentation of antigenic peptides to the direct MHC class I pathway via autophagy. We observed no difference in presentation of peptides derived from the viral EBNA1 protein following suppression of autophagy by knocking down Atg5 and Atg12. However, the same knock down treatment suppressed the presentation from ovalbumin. Fusing the aggregate-prone poly-glutamine (PolyQ) to the ovalbumin had no effect on antigen presentation via autophagy. Interestingly, fusing the EBNA1-derived gly-ala repeat (GAr) sequence to ovalbumin rendered the presentation Atg5/12 independent. We also demonstrate that the relative levels of protein expression did not affect autophagy-mediated antigen presentation. These data suggest a substrate-dependent presentation of antigenic peptides for the MHC class I pathway via autophagy and indicate that the GAr of the EBNA1 illustrates a novel virus-mediated mechanism for immune evasion of autophagy-dependent antigen presentation.

• Klíčová slova
• Autophagy, EBV-encoded EBNA1, MHC class I restricted antigen presentation, Protein aggregates,
• MeSH
• antigeny MeSH
• autofagie MeSH
• imunitní únik MeSH
• MHC antigeny I. třídy * MeSH
• MHC antigeny II. třídy metabolismus   MeSH
• ovalbumin MeSH
• prezentace antigenu * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny MeSH
• MHC antigeny I. třídy * MeSH
• MHC antigeny II. třídy MeSH
• ovalbumin MeSH

The knowledge of mechanisms of regulation of IL-10 production by B cells remains still very limited. We show here that highly purified mouse B cells stimulated with LPS produce significant levels of IL-10, but Bregs in our model do not express detectable level of either Foxp3 or GATA-3. Nevertheless, IL-10 production by B cells is regulated by cytokines. In activated B cells, IL-10 production was significantly enhanced by IFN-γ and decreased in the presence of IL-4 or TGF-β. These findings are in sharp contrast with the observations in T cells, where IL-10 production correlates with GATA-3 or FoxP3 expression, and the cytokines regulate IL-10 production in a reverse manner than in activated B cells. These results thus show that the production of IL-10 by Bregs is regulated by cytokines independently of the expression of GATA-3 and FoxP3, which is clearly different from GATA-3-dependent IL-10 production by activated Th2 cells and FoxP3 expression in IL-10-producing Tregs.

• Klíčová slova
• B cell, Cytokine, FoxP3, GATA-3, IL-10 production, T cell,
• MeSH
• aktivace lymfocytů imunologie   MeSH
• faktor 1 indukovatelný hypoxií - podjednotka alfa imunologie   MeSH
• forkhead transkripční faktory metabolismus   MeSH
• interferon gama imunologie   MeSH
• interleukin-10 biosyntéza   MeSH
• interleukin-4 imunologie   MeSH
• kultivované buňky MeSH
• lipopolysacharidy imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• regulační B-lymfocyty imunologie   MeSH
• regulační T-lymfocyty imunologie   MeSH
• Th2 buňky imunologie   MeSH
• transformující růstový faktor beta imunologie   MeSH
• transkripční faktor GATA3 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• faktor 1 indukovatelný hypoxií - podjednotka alfa MeSH
• forkhead transkripční faktory MeSH
• Foxp3 protein, mouse MeSH Prohlížeč
• Gata3 protein, mouse MeSH Prohlížeč
• Hif1a protein, mouse MeSH Prohlížeč
• IFNG protein, mouse MeSH Prohlížeč
• IL10 protein, mouse MeSH Prohlížeč
• Il4 protein, mouse MeSH Prohlížeč
• interferon gama MeSH
• interleukin-10 MeSH
• interleukin-4 MeSH
• lipopolysacharidy MeSH
• transformující růstový faktor beta MeSH
• transkripční faktor GATA3 MeSH

While autoimmune T cells are present in most individuals, only a minority of the population suffers from an autoimmune disease. To better appreciate the limits of T cell tolerance, we carried out experiments to determine how many autoimmune T cells are required to initiate an experimental autoimmune disease. Variable numbers of autoimmune OT-I T cells were transferred into RIP-OVA mice, which were injected with antigen-loaded DCs in a single footpad; this restricted T cell priming to a few OT-I T cells that are present in the draining popliteal lymph node. Using selective plane illumination microscopy (SPIM) we counted the number of OT-I T cells present in the popliteal lymph node at the time of priming. Analysis of our data suggests that a single autoimmune T cell cannot induce an experimental autoimmune disease, but a "quorum" of 2-5 autoimmune T cells clearly has this capacity.

• Klíčová slova
• Autoimmunity, T cell, Tolerance,
• MeSH
• autoimunita * MeSH
• autoimunitní nemoci imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• dendritické buňky imunologie   MeSH
• experimentální diabetes mellitus imunologie   MeSH
• imunologická tolerance MeSH
• lymfatické uzliny cytologie   imunologie   MeSH
• modely nemocí na zvířatech MeSH
• myši transgenní MeSH
• myši MeSH
• ovalbumin imunologie   MeSH
• převzatá imunita MeSH
• prezentace antigenu MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ovalbumin MeSH

Invariant natural killer T (iNKT) cells, CD1d restricted T cells, are involved in the immune responses against various infection agents. Here we describe their behavior during reactivation of human herpes simplex virus (HSV). iNKT cells exhibit only discrete changes, which however, reached statistically significant level due to the relatively large patient group. Higher percentage of iNKT cells express NKG2D. iNKT cells down-regulate NKG2A in a subset of patients. Finally, iNKT cells enhance their capacity to produce TNF-α. Our data suggests that iNKT cells are involved in the immune response against HSV and contribute mainly to its early, innate phase.

• MeSH
• aktivace lymfocytů MeSH
• aktivace viru MeSH
• antigeny CD1d genetika   imunologie   MeSH
• dospělí MeSH
• exprese genu MeSH
• herpes simplex imunologie   patologie   virologie   MeSH
• lektinové receptory NK-buněk - podrodina K genetika   imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• NKT buňky imunologie   patologie   MeSH
• přirozená imunita * MeSH
• senioři MeSH
• Simplexvirus fyziologie   MeSH
• TNF-alfa genetika   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD1d MeSH
• KLRK1 protein, human MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• TNF-alfa MeSH

Type 1 diabetes (T1D) is an autoimmune disease caused by T-cell mediated destruction of pancreatic beta cells. Recently, small cationic α-defensin molecules have been implicated in the pathogenesis of certain inflammatory and autoimmune diseases. The purpose of this study was to assess the α-defensin expression in patients with T1D and elucidate the cellular source of their production. Our results show that 30% of patients exhibit increased levels of α-defensin mRNAs in their capillary blood. Quantitative RT-PCR performed on FACS-sorted granulocytes identified CD15(dull)/CD14(weak) population as the cellular source of α-defensins. Surprisingly, this granulocyte subpopulation displayed augmentation of α-defensin expression in all T1D patients tested. The determination of cell surface markers, expression of cell-specific genes and confocal microscopy identified CD15(dull)/CD14(weak) cells as eosinophils. The presence of transcriptionally active eosinophils in diabetic patients suggests that eosinophils could be a part of an intricate innate immune cellular network involved in the development of diabetes.

• MeSH
• alfa-defensiny krev   genetika   imunologie   MeSH
• antigen Lewis X imunologie   MeSH
• autoimunita MeSH
• beta-buňky imunologie   metabolismus   patologie   MeSH
• diabetes mellitus 1. typu krev   genetika   imunologie   MeSH
• dítě MeSH
• dospělí MeSH
• eozinofily imunologie   metabolismus   patologie   MeSH
• exprese genu MeSH
• imunologická tolerance MeSH
• lidé MeSH
• lipopolysacharidové receptory imunologie   MeSH
• messenger RNA biosyntéza   MeSH
• mladiství MeSH
• peroxidasa krev   genetika   imunologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• předškolní dítě MeSH
• přirozená imunita MeSH
• průtoková cytometrie MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alfa-defensiny MeSH
• antigen Lewis X MeSH
• lipopolysacharidové receptory MeSH
• messenger RNA MeSH
• peroxidasa MeSH

Dendritic cells (DCs) play the key role in T-lymphocyte proliferation and induction of antitumour response. The mixed leukocyte reaction (MLR) of T-lymphocytes and DCs is essential instrument for immunological mechanisms studies. Conventionally used method for determination of T-lymphocytes proliferation, (3)H-thymidine incorporation, provides only general information. The method of flow cytometry and differential gating seems to be more suitable for quantitative and qualitative analysis of T-lymphocyte proliferation. It is based on time limited acquisition of events and on its distribution according to forward and side scatter values. We decided to compare these two methods and determine mutual correlation and compatibility. Eleven patients were studied and in all cases DCs promoted the survival and proliferation of both CD4 and CD8 lymphocytes. Both methods retained consistency with regard to survival and proliferation of CD4/CD8 lymphocytes. However, the correlation of these methods was not convincing. Therefore, both these methods might be used for evaluation of MLR, but each of them gives specific and complementary information.

• MeSH
• CD antigeny imunologie   metabolismus   MeSH
• CD4-pozitivní T-lymfocyty imunologie   metabolismus   patologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   metabolismus   patologie   MeSH
• chronická lymfatická leukemie imunologie   patologie   MeSH
• dendritické buňky imunologie   patologie   MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipopolysacharidové receptory imunologie   metabolismus   MeSH
• lymfocyty imunologie   metabolismus   patologie   MeSH
• monocyty imunologie   metabolismus   patologie   MeSH
• proliferace buněk * MeSH
• průtoková cytometrie metody   MeSH
• receptory transferinu imunologie   metabolismus   MeSH
• reprodukovatelnost výsledků MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• test smíšené lymfocytární kultury metody   MeSH
• thymidin metabolismus   MeSH
• tritium MeSH
• viabilita buněk imunologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CD antigeny MeSH
• CD71 antigen MeSH Prohlížeč
• lipopolysacharidové receptory MeSH
• receptory transferinu MeSH
• thymidin MeSH
• tritium MeSH

Invariant natural killer T (iNKT) cells play an important role in the immune response against various infectious agents. In this study we investigated their role in human defense against the varicella zoster virus. We observed decreased numbers of iNKT cells in patients who failed to control latent varicella zoster virus infection, e.g. underwent several reactivations of the virus. The residual population of iNKT cells expressed significantly higher levels of inhibitory receptor CD158a that was further up-regulated in the course of acute viral infection. Both of these abnormalities might contribute to impaired control of varicella zoster virus in human.

• MeSH
• aktivace lymfocytů imunologie   MeSH
• buňky NK imunologie   metabolismus   patologie   MeSH
• CD antigeny metabolismus   MeSH
• cytokiny metabolismus   MeSH
• herpes zoster imunologie   patologie   MeSH
• lektinové receptory NK-buněk - podrodina K metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• NKT buňky imunologie   metabolismus   patologie   MeSH
• počet lymfocytů MeSH
• podskupiny lymfocytů imunologie   metabolismus   patologie   MeSH
• receptory KIR2DL1 metabolismus   MeSH
• T-lymfocyty imunologie   metabolismus   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CD antigeny MeSH
• cytokiny MeSH
• KIR2DL1 protein, human MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• receptory KIR2DL1 MeSH

Regulatory T cells (Tregs) are critical regulators of autoimmune diseases, including type 1 diabetes mellitus. It is hypothesised that Tregs' function can be influenced by changes in the expression of specific microRNAs (miRNAs). Thus, we performed miRNAs profiling in a population of Tregs separated from peripheral blood of five type 1 diabetic patients and six healthy donors. For more detailed molecular characterisation of Tregs, we additionally compared miRNAs expression profiles of Tregs and conventional T cells. Tregs were isolated according to CD3+, CD4+, CD25(hi)+ and CD127- by flow cytometry, and miRNA expression profiling was performed using TaqMan Array Human MicroRNA Panel-1 (384-well low density array). In Tregs of diabetic patients we found significantly increased expression of miRNA-510 (p=0.05) and decreased expression of both miRNA-342 (p<0.0001) and miRNA-191 (p=0.0079). When comparing Tregs and T cells, we revealed that Tregs had significant higher expression of miRNA-146a and lower expression of eight specific miRNAs (20b, 31, 99a, 100, 125b, 151, 335, and 365). To our knowledge, this is the first study demonstrating changes in miRNA expression profiles occurring in Tregs of T1D patients and a miRNAs signature of adult Tregs.

• MeSH
• diabetes mellitus 1. typu krev   genetika   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikro RNA genetika   MeSH
• mladý dospělý MeSH
• regulační T-lymfocyty metabolismus   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody   MeSH
• shluková analýza MeSH
• stanovení celkové genové exprese * MeSH
• T-lymfocyty metabolismus   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
• práce podpořená grantem MeSH
• Názvy látek
• mikro RNA MeSH
• MIRN191 microRNA, human MeSH Prohlížeč
• MIRN342 microRNA, human MeSH Prohlížeč
• MIRN510 microRNA, human MeSH Prohlížeč

We analyzed the frequency and absolute numbers of circulating myeloid and plasmacytoid DCs in peripheral blood and evaluated their maturation status to test the hypothesis that significant physical stress to the body might induce measurable changes in DCs subsets, phenotype and function, which would complete existing knowledge about the response of the cellular immune system to an acute exercise in top sportsmen. We evaluated the heart rate and draw blood samples before and after the physical load in 18 profesional ice-hockey players. We observed an increase in leukocytes numbers with a predominant increase in the population of DCs and lymphocytes after exercise. Both myeloid and plasmacytoid DCs increased significantly. We found a correlation between the increase of peripheral blood DCs and serum epinephrine and norepinephrine levels. Increase in peripheral blood DCs also correlates with the extent of heart rate elevation during exercise.

• MeSH
• adrenalin krev   MeSH
• CD antigeny metabolismus   MeSH
• cvičení fyziologie   MeSH
• dendritické buňky cytologie   metabolismus   MeSH
• dospělí MeSH
• hokej fyziologie   MeSH
• lidé MeSH
• noradrenalin krev   MeSH
• počet leukocytů MeSH
• počet lymfocytů MeSH
• srdeční frekvence fyziologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adrenalin MeSH
• CD antigeny MeSH
• noradrenalin MeSH

Type 1 diabetes mellitus (T1D) and multiple sclerosis (MS) are organ-specific autoimmune diseases leading to an attack of auto-aggressive lymphocytes against the pancreatic beta-cells and central nervous system, respectively. Using four-colour flow cytometry, T-lymphocyte populations having an important function in autoimmune processes were analyzed. T-regulatory cells (Treg) CD4(+)CD25(+)CD127(low), T-suppressor cells (Ts) CD8(+)CD28(-), activated helper CD4(+)CD25(+)CD127(+) and cytotoxic CD8(+)CD25(+) T-cells and also naive CD4(+)CD45RA(+) and memory T-cells CD4(+)CD45RO(+) were compared in the group of patients with T1D (n=30), MS (n=31) and in the group of healthy controls (n=29). Significant differences in Ts cells, activated helper and cytotoxic cells and also memory T-cells were recognized in the group of T1D patients compared to healthy controls. Ts population was significantly lowered in MS patients as well. However, no significant differences were noticed in Treg population. The observed data demonstrate significant differences among patients with T1D and MS in comparison to healthy individuals.

• MeSH
• antigeny CD28 imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• diabetes mellitus 1. typu imunologie   MeSH
• dospělí MeSH
• imunologická paměť imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• počet lymfocytů * MeSH
• průtoková cytometrie metody   MeSH
• relabující-remitující roztroušená skleróza imunologie   MeSH
• T-lymfocyty - podskupiny cytologie   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD28 MeSH