CD4 and CD8 mark helper and cytotoxic T cell lineages, respectively, and serve as coreceptors for MHC-restricted TCR recognition. How coreceptor expression is matched with TCR specificity is central to understanding CD4/CD8 lineage choice, but visualising coreceptor gene activity in individual selection intermediates has been technically challenging. It therefore remains unclear whether the sequence of coreceptor gene expression in selection intermediates follows a stereotypic pattern, or is responsive to signaling. Here we use single cell RNA sequencing (scRNA-seq) to classify mouse thymocyte selection intermediates by coreceptor gene expression. In the unperturbed thymus, Cd4+Cd8a- selection intermediates appear before Cd4-Cd8a+ selection intermediates, but the timing of these subsets is flexible according to the strength of TCR signals. Our data show that selection intermediates discriminate MHC class prior to the loss of coreceptor expression and suggest a model where signal strength informs the timing of coreceptor gene activity and ultimately CD4/CD8 lineage choice.

• MeSH
• aktivace lymfocytů genetika   MeSH
• analýza hlavních komponent MeSH
• buněčná diferenciace imunologie   MeSH
• buněčný rodokmen imunologie   MeSH
• CD4-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• cytokiny metabolismus   MeSH
• DNA vazebné proteiny metabolismus   MeSH
• histokompatibilní antigeny metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• protein PEBP2A3 metabolismus   MeSH
• receptory antigenů T-buněk metabolismus   MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• thymus cytologie   imunologie   MeSH
• transkripční faktory metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• cytokiny MeSH
• DNA vazebné proteiny MeSH
• histokompatibilní antigeny MeSH
• messenger RNA MeSH
• protein PEBP2A3 MeSH
• receptory antigenů T-buněk MeSH
• Runx3 protein, mouse MeSH Prohlížeč
• transkripční faktory MeSH
• Zbtb7b protein, mouse MeSH Prohlížeč

The organizational integrity of the adaptive immune system is determined by functionally discrete subsets of CD4+ T cells, but it has remained unclear to what extent lineage choice is influenced by clonotypically expressed T-cell receptors (TCRs). To address this issue, we used a high-throughput approach to profile the αβ TCR repertoires of human naive and effector/memory CD4+ T-cell subsets, irrespective of antigen specificity. Highly conserved physicochemical and recombinatorial features were encoded on a subset-specific basis in the effector/memory compartment. Clonal tracking further identified forbidden and permitted transition pathways, mapping effector/memory subsets related by interconversion or ontogeny. Public sequences were largely confined to particular effector/memory subsets, including regulatory T cells (Tregs), which also displayed hardwired repertoire features in the naive compartment. Accordingly, these cumulative repertoire portraits establish a link between clonotype fate decisions in the complex world of CD4+ T cells and the intrinsic properties of somatically rearranged TCRs.

• Klíčová slova
• CDR3 properties, TCR repertoire, helper CD4+ subsets, human, immunology, inflammation, plasticity of CD4+ subsets,
• MeSH
• buněčný rodokmen imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• lidé MeSH
• receptory antigenů T-buněk alfa-beta imunologie   MeSH
• T-lymfocyty - podskupiny imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• receptory antigenů T-buněk alfa-beta MeSH

Medullary thymic epithelial cell (mTEC)-restricted expression of autoimmune regulator (Aire) is essential for establishment of immune tolerance. Recently, Aire was also shown to be expressed in cells of hematopietic and reproductive lineages. Thus, the generation of Airefl/fl mouse strain enables the investigation of the cell-specific function of Aire.

• Klíčová slova
• Aire, Conditional knockout, Immune tolerance, Medullary thymic epithelial cells,
• MeSH
• antigen prezentující buňky patologie   MeSH
• autoimunitní polyglandulární syndromy genetika   imunologie   patologie   MeSH
• buněčný rodokmen genetika   imunologie   MeSH
• imunologická tolerance genetika   MeSH
• lidé MeSH
• myši knockoutované MeSH
• myši transgenní MeSH
• myši MeSH
• protein AIRE MeSH
• rozmnožování genetika   imunologie   MeSH
• transkripční faktory genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH
• Názvy látek
• transkripční faktory MeSH

• MeSH
• buněčná diferenciace genetika   imunologie   MeSH
• buněčný rodokmen genetika   imunologie   MeSH
• fenotyp MeSH
• hematopoéza genetika   MeSH
• imunofenotypizace metody   MeSH
• leukemie genetika   imunologie   patologie   MeSH
• lidé MeSH
• myeloidní buňky fyziologie   MeSH
• prekurzory T-buněk imunologie   patologie   MeSH
• sekvenování exomu MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

Mucosal-associated invariant T (MAIT) cells contain two main subpopulations, CD8(+) and double-negative (DN) cells. The first reports suggested that subpopulations of MAIT cells have similar phenotype and function. Recent works, however, demonstrate that the subpopulations have different ontogenesis and are differentially affected by xenobiotic treatment. In this work, we re-examined the possible differences between subpopulations of MAIT cells. We demonstrate that the main subpopulations of MAIT cells (CD8 and DN) are relatively uniform in terms of both phenotype and function. Both populations are memory/activated, tissue-homing and pro-inflammatory. CD8(+) MAIT cells are better equipped for pro-inflammatory functions as they express higher levels of CD16 and NKG2D, produce more pro-inflammatory cytokines (TNF-α and IFN-γ) and have higher cytotoxic potential (contain more granzyme B and express higher levels of CD107A upon stimulation). Our study contributes to the understanding of the heterogeneity of MAIT cell population.

• MeSH
• biologické markery metabolismus   MeSH
• buněčný rodokmen imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• dospělí MeSH
• exprese genu MeSH
• GPI-vázané proteiny genetika   imunologie   MeSH
• granzymy genetika   imunologie   MeSH
• imunofenotypizace MeSH
• imunologická paměť * MeSH
• interferon gama genetika   imunologie   MeSH
• lektinové receptory NK-buněk - podrodina K genetika   imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• MAIT buňky cytologie   imunologie   MeSH
• membránový protein 1 asociovaný s lyzozomy genetika   imunologie   MeSH
• receptory IgG genetika   imunologie   MeSH
• TNF-alfa genetika   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• FCGR3B protein, human MeSH Prohlížeč
• GPI-vázané proteiny MeSH
• granzymy MeSH
• GZMB protein, human MeSH Prohlížeč
• IFNG protein, human MeSH Prohlížeč
• interferon gama MeSH
• KLRK1 protein, human MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• membránový protein 1 asociovaný s lyzozomy MeSH
• receptory IgG MeSH
• TNF-alfa MeSH

Tick-borne encephalitis virus (TBEV) is a human-pathogenic flavivirus that is endemic in large parts of Europe and Asia and causes severe neuroinvasive illness. A formalin-inactivated vaccine induces strong neutralizing antibody responses and confers protection from TBE disease. CD4+ T cell responses are essential for neutralizing antibody production, but data on the functionalities of TBEV-specific CD4+ T cells in response to vaccination or infection are lacking. This study provides a comprehensive analysis of the cytokine patterns of CD4+ T cell responses in 20 humans after TBE vaccination in comparison to those in 18 patients with TBEV infection. Specifically, Th1-specific cytokines (IFN-γ, IL-2, TNF-α), CD40 ligand and the Th1 lineage-specifying transcription factor Tbet were determined upon stimulation with peptides covering the TBEV structural proteins contained in the vaccine (C-capsid, prM/M-membrane and E-envelope). We show that TBEV-specific CD4+ T cell responses are polyfunctional, but the cytokine patterns after vaccination differed from those after infection. TBE vaccine responses were characterized by lower IFN-γ responses and high proportions of TNF-α+IL-2+ cells. In vaccine-induced responses-consistent with the reduced IFN-γ expression patterns-less than 50% of TBEV peptides were detected by IFN-γ+ cells as compared to 96% detected by IL-2+ cells, indicating that the single use of IFN-γ as a read-out strongly underestimates the magnitude and breadth of such responses. The results provide important insights into the functionalities of CD4+ T cells that coordinate vaccine responses and have direct implications for future studies that address epitope specificity and breadth of these responses.

• MeSH
• buněčný rodokmen imunologie   MeSH
• CD4-pozitivní T-lymfocyty účinky léků   imunologie   MeSH
• cytokiny biosyntéza   imunologie   MeSH
• dospělí MeSH
• epitopy imunologie   MeSH
• klíšťová encefalitida imunologie   prevence a kontrola   virologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• protilátky virové krev   imunologie   MeSH
• senioři MeSH
• T-lymfocyty pomocné-indukující účinky léků   imunologie   MeSH
• vakcinace * MeSH
• virové vakcíny aplikace a dávkování   imunologie   MeSH
• viry klíšťové encefalitidy imunologie   patogenita   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny MeSH
• epitopy MeSH
• protilátky virové MeSH
• virové vakcíny MeSH

Porcine γδ T cells have two levels of TCRγδ expression. Whereas TCRγδ(med) cells are mostly CD2(+)CD8(-) and CD2(+)CD8(+), TCRγδ(hi) cells are highly enriched for CD2(-)CD8(-). This distribution is independent of bacterial colonization and it is already established in the thymus prior to export of γδ cells to the periphery. Sorting and cultivation experiments revealed that CD2(-)CD8(-) γδ cells are unable to acquire CD2 and CD8, whereas CD2(+) subsets can gain or loose CD8. There is also differential susceptibility for proliferation between CD2(+) and CD2(-) γδ cells. Although CD2(-)CD8(-) almost do not proliferate, proliferation of CD2(+)CD8(-) and CD2(+)CD8(+) is substantial. Population of CD2(-) γδ cells is also absent in CD1(+) immature thymocytes. Additionally, subpopulations of CD2(+) and CD2(-) γδ cells in the thymus differ in expression of auxiliary surface molecules such as CD25, CD45RA/RC, and MHC class II. Moreover, TCRγδ(hi) cells can generate TCRγδ(med) cells but never the opposite. The only exception is the thymus, where a few TCRγδ(med) cells can be induced to TCRγδ(hi) but only under IL-2 influence. The repertoire of TCRδ is polyclonal in all subsets, indicating that there is the same extent of diversification and equal capability of immune responses. Results collectively indicate that CD2 expression determines two lineages of γδ cells that differ in many aspects. Because CD2(-) γδ cells are missing in the blood of humans and mice but are obvious in other members of γδ-high species such as ruminants and birds, our findings support the idea that circulating CD2(-) γδ T cells are a specific lineage.

• MeSH
• antigeny CD1 genetika   imunologie   MeSH
• antigeny CD2 genetika   imunologie   MeSH
• antigeny CD8 genetika   imunologie   MeSH
• buněčná diferenciace imunologie   MeSH
• buněčný rodokmen imunologie   MeSH
• exprese genu MeSH
• interleukin-2 imunologie   MeSH
• lidé MeSH
• myši MeSH
• prasata MeSH
• proliferace buněk MeSH
• receptory antigenů T-buněk gama-delta genetika   imunologie   MeSH
• T-lymfocyty - podskupiny cytologie   imunologie   MeSH
• T-lymfocyty cytologie   imunologie   MeSH
• thymocyty cytologie   imunologie   MeSH
• thymus cytologie   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD1 MeSH
• antigeny CD2 MeSH
• antigeny CD8 MeSH
• interleukin-2 MeSH
• receptory antigenů T-buněk gama-delta MeSH

Based on studies of sheep, ileal Peyer's patches (IPP) have been regarded as a type of primary lymphoid tissue similar to the bursa of Fabricius in chicken. Because bursectomy results in B cell deficiency, we wondered whether resection of the IPP of piglets would have a similar effect. Comparison of IPP-resected, surgical shams and untreated germ-free piglets, all of which were later colonized with a defined commensal flora, demonstrated that resection of the IPP did not alter the level and phenotype of B and T cells in lymphoid tissues and the blood 10 wk after surgery. Additionally, colonization of IPP caused a shift from the fetal type of lymphocyte distribution to the adult type that is characterized by prevalence of B cells, with many of them representing IgA(+) switched B cells or displaying a more mature CD2(-)CD21(+) and CD2(-)CD21(-) phenotype. Moreover, colonization leads to appearance of effector CD4(+)CD8(+) αβ T helper and CD2(+)CD8(-) γδ T cells. Comparison of germ-free with colonized pigs and experiments utilizing surgical transposition of jejunal Peyer's patch into terminal ileum or construction of isolated ileal loops indicated that lymphocyte development in IPP is dependent on colonization. Although our studies confirmed higher mitotic and apoptotic rates in IPP, they failed to identify any cell populations that resemble developing B lineage cells in the bone marrow. These results indicate that porcine IPP are not required for systemic B cell generation or maintenance, but they are secondary lymphoid tissue that appears important in immune responses to colonizing bacteria.

• MeSH
• buněčná diferenciace imunologie   MeSH
• buněčný rodokmen imunologie   MeSH
• gnotobiologické modely MeSH
• ileum cytologie   imunologie   chirurgie   MeSH
• lymfopoéza imunologie   MeSH
• novorozená zvířata MeSH
• Peyerovy pláty cytologie   imunologie   chirurgie   MeSH
• počet lymfocytů MeSH
• podskupiny B-lymfocytů cytologie   imunologie   metabolismus   MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• srovnávací studie MeSH

Cancer microenvironment is increasingly recognized as an important factor affecting cancer onset and progression. Since Wirchow reported in 1863 that tumors contain inflammatory cells, the field shifted significantly forward, and immune cells residing in tumors appear to be attractive targets of cancer therapies. For some methods, such as stem/progenitor cell isolation from both cancer and healthy tissues, removal of contaminating immune cells is crucial to achieve consistent, reproducible and accurate results. Despite current methods of lineage negative selection accounts for removal of over 99 % of immune cells from stem/progenitor cell isolates, the vast majority of lineage antibody cocktails retain basophils, dendritic cells, and mast cells. Here we discuss the ability of the most commonly used lineage markers to bind to the plasma membrane of mast cells and/or basophils, and suggest alternatives, which may be used for negative selection of these cellular populations. Both, mast cells and basophils, were shown to participate actively in cancer-associated angiogenesis, tissue remodeling and recruitment of other immune cell types, including eosinophils, B cells, memory T cells and Treg cells. In turn, tumor-derived peptides and chemotactic factors are known to recruit and activate mast cells in neoplasias, resulting in altered tumor progression. Repeated findings of CD34+ populations of mast cells and basophils further highlight necessity of their separation from stem/progenitor cell isolates in both, preclinical experiments and clinical praxis.

• MeSH
• antigeny povrchové biosyntéza   imunologie   MeSH
• bazofily účinky léků   imunologie   metabolismus   patologie   MeSH
• bibliometrie MeSH
• buněčný rodokmen imunologie   MeSH
• databáze faktografické statistika a číselné údaje   MeSH
• kmenové buňky imunologie   patologie   MeSH
• lidé MeSH
• mastocyty účinky léků   imunologie   metabolismus   patologie   MeSH
• molekulární struktura MeSH
• nádory * farmakoterapie   imunologie   patologie   MeSH
• protinádorové látky chemie   farmakologie   terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• antigeny povrchové MeSH
• protinádorové látky MeSH

Developmental pathways of gammadelta T cells are still unknown, largely because of the absence of recognized lineage-specific surface markers other than the TCR. We have shown that porcine gammadelta thymocytes can be divided into 12 subsets of the following two major groups: 1) CD4(-) gammadelta thymocytes that can be further subdivided according to their CD2/CD8alphaalpha phenotype, and 2) CD4(+) gammadelta thymocytes that are always CD1(+)CD2(+)CD8alphabeta(+) and have no counterpart in the periphery. In this study, we have analyzed gammadelta thymocyte subsets with respect to behavior during cultivation, cell cycle status, and lymphocyte-specific transcripts. The group of CD4(-) gammadelta thymocytes gives rise to all gammadelta T cells found in the periphery. Proliferating CD2(+)CD8(-)CD1(+)CD45RC(-) gammadelta thymocytes are a common precursor of this group. These precursors differentiate into CD2(+)CD8alphaalpha(+), CD2(+)CD8(-), and CD2(-)CD8(-) gammadelta T cell subsets, which subsequently mature by loss of CD1 and by eventual gain of CD45RC expression. In contrast, the group of CD4(+) gammadelta thymocytes represents transient and independent subsets that are never exported from thymus as TCRgammadelta(+) T cells. In accordance with the following findings, we propose that CD4(+)CD8alphabeta(+) gammadelta thymocytes extinguish their TCRgammadelta expression and differentiate along the alphabeta T cell lineage program: 1) CD4(+) gammadelta thymocytes are actively dividing; 2) CD4(+) gammadelta thymocytes do not die, although their numbers decreased with prolonged cultivation; 3) CD4(+) gammadelta thymocytes express transcripts for RAG-1, TdT, and TCRbeta; and 4) CD4(+) gammadelta thymocytes are able to alter their phenotype to TCRalphabeta(+) thymocytes under appropriate culture conditions.

• MeSH
• antigeny CD1 metabolismus   MeSH
• antigeny CD2 metabolismus   MeSH
• antigeny CD45 imunologie   metabolismus   MeSH
• apoptóza MeSH
• buněčná diferenciace imunologie   MeSH
• buněčný rodokmen imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   metabolismus   MeSH
• CD8-pozitivní T-lymfocyty imunologie   metabolismus   MeSH
• fenotyp MeSH
• genetická transkripce genetika   MeSH
• kultivované buňky MeSH
• pohyb buněk MeSH
• prasata imunologie   MeSH
• receptory antigenů T-buněk gama-delta genetika   imunologie   MeSH
• thymus cytologie   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD1 MeSH
• antigeny CD2 MeSH
• antigeny CD45 MeSH
• receptory antigenů T-buněk gama-delta MeSH