γδ T lymfocyty představují menšinovou populaci T lymfocytů, která se v základu liší konstrukcí TCR receptoru. Unikátní vlastnosti γδ TCR dávají pak těmto buňkám jedinečné efektorové funkce a specifickou roli (nejen) v protinádorové imunitní odpovědi. V tomto článku popisujeme základní charakteristiku těchto buněk ve vztahu k onkologickým onemocněním. V experimentální části je pak provedena exploratorní analýza zastoupení γδ T lymfocytů v běžné populaci a srovnání těchto hodnot s hodnotami pacientů s melanomem a karcinomem prsu. Medián procentuálního zastoupení γδ ze všech lymfocytů byl 2,9 % (interkvartilové rozpětí – IQR 1,7– 4 %). Medián absolutních počtů γδ buněk v litru krve byl 5,05 × 107 (IQR 2,9– 7,84 × 107). Medián procentuálního zastoupení γδ buněk mezi T lymfocyty byl 3,9 % (IQR 2,3– 5,6 %). V referenční populaci nebyla prokázána závislost kvantitativních parametrů γδ buněk na pohlaví či věku. Dále proběhla detailní imunofenotypizace popisující zastoupení paměťových subpopulací (pomocí značení CD45RO a CD27) a výskyt povrchových markerů HLA‑Dr, CD69, CD25, CD28, CCR7, CTLA‑ 4, ICOS, PD‑ 1L a PD‑ 1 mezi γδ T lymfocyty u kontrol a pacientek s karcinomem prsu. Z této analýzy je patrné, že γδ buňky netvoří uniformní populaci, ale mohou se ve svých povrchových markerech lišit, stejně jako se pak liší v efektorových funkcích.

γδ T cells present a minor population of the T cell family which basically differs in construction of their T cell receptor (TCR). Thanks to the features of γδ TCR, these cells can acquire unique effector functions and play a specific role (not only) in anti‑tumor immune response. In this article, we describe the basic characteristics of this cell population and their connection to cancer. In the experimental part we performed exploratory analysis of circulating γδ T cells in reference population and comparison with melanoma and breast carcinoma patients. The median percentage of γδ T cells from all lymphocytes was 2.9% (interquartile range – IQR 1.7– 4%). The median absolute numbers of γδ cells per liter of blood was 5.05 × 107 (IQR 2.9– 7.84 × 107). The median percentage of γδ cells between all CD3 T cells was 3.9% (IQR 2.3– 5.6%). No correlation between γδ T cells levels and gender or age was observed in reference population. Detailed immunophenotyping was also conducted describing representation of memory subsets (using CD45RO and CD27 markers) and presence of surface markers HLA‑Dr, CD69, CD25, CD28, CCR7, CTLA‑ 4, ICOS, PD‑ 1L and PD‑ 1 between γδ T cells of the controls and breast carcinoma patients. From this analysis, it is evident that γδ T cells do not represent a uniform population but they differ in surface markers as well as in their effector functions.

• Klíčová slova
• γδ T-lymfocyty,
• MeSH
• dospělí MeSH
• imunitní systém MeSH
• imunofenotypizace MeSH
• imunoterapie adoptivní metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• melanom krev   metabolismus   MeSH
• nádory kůže krev   metabolismus   MeSH
• nádory prsu krev   metabolismus   MeSH
• nádory * farmakoterapie   imunologie   MeSH
• průtoková cytometrie statistika a číselné údaje   MeSH
• receptory antigenů T-buněk gama-delta * fyziologie   imunologie   terapeutické užití   MeSH
• rozložení podle pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• statistika jako téma MeSH
• studie případů a kontrol MeSH
• T-lymfocyty * 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
• kazuistiky MeSH
• práce podpořená grantem MeSH

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

Psoriasis vulgaris (PV) is a chronic, recurrent inflammatory dermatosis mediated by aberrantly activated immune cells. The role of the innate-like T cells, particularly gammadelta T (γδT) cells and MR1-restricted T lymphocytes, is incompletely explored, mainly through animal models, or by use of surrogate lineage markers, respectively. Here, we used case-control settings, multiparameter flow cytometry, 5-OP-RU-loaded MR1-tetramers, Luminex technology and targeted qRT-PCR to dissect the cellular and transcriptional landscape of γδ and MR1-restricted blood T cells in untreated PV cases (n=21, 22 matched controls). High interpersonal differences in cell composition were observed, fueling transcriptional variability at healthy baseline. A minor subset of canonical CD4+CD8+MR1-tet+TCRVα7.2+ and CD4+CD8-MR1-tet+TCRVα7.2+ T cells was the most significantly underrepresented community in male PV individuals, whereas Vδ2+ γδ T cells expressing high levels of TCR and Vδ1-δ2- γδ T cells expressing intermediate levels of TCR were selectively enriched in affected males, partly reflecting disease severity. Our findings highlight a formerly unappreciated skewing of human circulating MAIT and γδ cytomes during PV, and reveal their compositional changes in relation to sex, CMV exposure, serum cytokine content, BMI, and inflammatory burden. Complementing numerical alterations, we finally show that flow-sorted, MAIT and γδ populations exhibit divergent transcriptional changes in mild type I psoriasis, consisting of differential bulk expression for signatures of cytotoxicity/type-1 immunity (EOMES, RUNX3, IL18R), type-3 immunity (RORC, CCR6), and T cell innateness (ZBTB16).

• MeSH
• buněčná diferenciace MeSH
• cytotoxicita imunologická MeSH
• dospělí MeSH
• histokompatibilita - antigeny třídy I metabolismus   MeSH
• krevní oběh MeSH
• lidé středního věku MeSH
• lidé MeSH
• MAIT buňky imunologie   MeSH
• mladý dospělý MeSH
• přirozená imunita MeSH
• protein promyelocytické leukemie s motivem zinkového prstu genetika   metabolismus   MeSH
• psoriáza imunologie   MeSH
• receptory antigenů T-buněk gama-delta metabolismus   MeSH
• T-lymfocyty imunologie   MeSH
• Th1 buňky imunologie   MeSH
• vedlejší histokompatibilní antigeny 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

The mammalian body possesses remarkable adaptability to cold exposure, involving intricate adjustments in cellular metabolism, ultimately leading to thermogenesis. However, cold-induced stress can impact immune response, primarily through noradrenaline-mediated pathways. In our study, we utilized a rat model subjected to short-term or long-term mild cold exposure to investigate systemic immune response during the cold acclimation. To provide human relevance, we included a group of regular cold swimmers in our study. Our research revealed complex relationship between cold exposure, neural signaling, immune response, and thermogenic regulation. One-day cold exposure triggered stress response, including cytokine production in white adipose tissue, subsequently activating brown adipose tissue, and inducing thermogenesis. We further studied systemic immune response, including the proportion of leukocytes and cytokines production. Interestingly, γδ T cells emerged as possible regulators in the broader systemic response, suggesting their possible contribution in the dynamic process of cold adaptation. We employed RNA-seq to gain further insights into the mechanisms by which γδ T cells participate in the response to cold. Additionally, we challenged rats exposed to cold with the Toll-like receptor 2 agonist, showing significant modulation of immune response. These findings significantly contribute to understanding of the physiological acclimation that occur in response to cold exposure.

• MeSH
• aklimatizace imunologie   MeSH
• cytokiny metabolismus   MeSH
• hnědá tuková tkáň imunologie   metabolismus   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• nízká teplota * MeSH
• receptory antigenů T-buněk gama-delta imunologie   metabolismus   MeSH
• T-lymfocyty imunologie   MeSH
• termogeneze imunologie   MeSH
• toll-like receptor 2 * metabolismus   genetika   imunologie   MeSH
• zánět * imunologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Severe combined immunodeficiencies (SCID) comprise a group of genetic diseases characterized by abrogated development of T lymphocytes. In some case reports of atypical SCID patients elevated proportions of γδ T lymphocytes have been reported. However, it is unknown whether these γδ T cells modulate or reflect the patient's clinical phenotype. We investigated the frequency of elevated γδ T cell proportions and associations with clinical disease manifestations in a cohort of 76 atypical SCID patients. Increased proportions of γδ T lymphocytes were present in approximately 60% of these patients. Furthermore, we identified positive correlations between elevated proportions of γδ T cells and the occurrence of CMV infections and autoimmune cytopenias. We discuss that CMV infections might trigger an expansion of γδ T lymphocytes, which could drive the development of autoimmune cytopenias. We advocate that atypical SCID patients should be screened for elevated proportions of γδ T lymphocytes, CMV infection and autoimmune cytopenias.

• MeSH
• cytomegalovirové infekce imunologie   MeSH
• intraepiteliální lymfocyty imunologie   MeSH
• krevní nemoci imunologie   MeSH
• lidé MeSH
• počet lymfocytů MeSH
• těžká kombinovaná imunodeficience imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH

Lymphocytes represent the key antigen-specific leukocyte subpopulation. Despite their importance in mounting an immune response, an unbiased description of proteins expressed by chicken lymphocytes has not been presented. In this study, we therefore intravenously infected chickens with Salmonella Enteritidis, sorted CD4, CD8 and γδ T-lymphocytes from the spleen by flow cytometry and determined the proteome of each population by LC-MS/MS. CD4 T-lymphocyte characteristic proteins included ubiquitin SUMO-like domain and BAR domain containing proteins. CD8 T-lymphocyte specific proteins were characterized by purine ribonucleoside triphosphate binding and were involved in cell differentiation, cell activation and regulation of programmed cell death. γδ T-lymphocyte specific proteins exhibited enrichment of small GTPase of Rab type and GTP binding. Following infection, inducible proteins in CD4 lymphocytes included ribosomal proteins and downregulated proteins localized to the lysosome. CD8 T-lymphocytes induced MCM complex proteins, proteins required for DNA replication and machinery for protein processing in the endoplasmic reticulum. Proteins inducible in γδ T-lymphocytes belonged to immune system response, oxidative phosphorylation and the spliceosome. In this study, we predicted the likely events in lymphocyte response to systemic bacterial infection and identified proteins which can be used as markers specific for each lymphocyte subpopulation.

• MeSH
• CD4-pozitivní T-lymfocyty imunologie   metabolismus   mikrobiologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   metabolismus   mikrobiologie   MeSH
• intraepiteliální lymfocyty imunologie   metabolismus   mikrobiologie   MeSH
• kur domácí imunologie   metabolismus   MeSH
• nemoci drůbeže imunologie   metabolismus   mikrobiologie   prevence a kontrola   MeSH
• Salmonella enteritidis imunologie   metabolismus   MeSH
• salmonelové vakcíny imunologie   MeSH
• salmonelóza imunologie   metabolismus   mikrobiologie   prevence a kontrola   MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

γδ T cells are intensively studied because their function in infection, allergy, autoimmune disease, cancer and post-transplant period is not yet fully understood. PCR-based techniques were established to study the γ variable (Vγ) and δ variable (Vδ) gene families. PCR product evaluation is routinely carried out by Southern blot analysis or the third complementarity-determining region spectratyping, but a fast and simple assessment of Vγ and Vδ gene family expression is missing. The aim of our study was to test capillary electrophoresis as a potential method for evaluating the composition of the γδ T-cell population. This report provides optimized PCR conditions for γδ T-cell receptor amplification. Further, it describes the utilization of capillary electrophoresis in the Agilent 2100 Bioanalyzer to evaluate the relative expression of Vγ and Vδ gene families after their amplification. An application of the methodology to peripheral blood mononuclear cell samples from patients during haemato-oncological treatment is shown. The described methodology is fast and simple to operate and is therefore suitable as a first screening of the γδ T-cell population composition in tissues of interest.

• MeSH
• časové faktory MeSH
• dospělí MeSH
• elektroforéza kapilární metody   MeSH
• hematologické nádory genetika   patologie   MeSH
• leukocyty mononukleární cytologie   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• multigenová rodina * MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• receptory antigenů T-buněk gama-delta genetika   metabolismus   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody   MeSH
• studie případů a kontrol MeSH
• T-lymfocyty cytologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Despite increasing interest in γδ T cells and their non-classical behaviour, most studies focus on animals with low numbers of circulating γδ T cells, such as mice and humans. Arguably, γδ T cell functions might be more prominent in chickens where these cells form a higher proportion of the circulatory T cell compartment. The TCR repertoire defines different subsets of γδ T cells, and such analysis is facilitated by well-annotated TCR loci. γδ T cells are considered at the cusp of innate and adaptive immunity but most functions have been identified in γδ low species. A deeper understanding of TCR repertoire biology in γδ high and γδ low animals is critical for defining the evolution of the function of γδ T cells. Repertoire dynamics will reveal populations that can be classified as innate-like or adaptive-like as well as those that straddle this definition. RESULTS: Here, a recent discrepancy in the structure of the chicken TCR gamma locus is resolved, demonstrating that tandem duplication events have shaped the evolution of this locus. Importantly, repertoire sequencing revealed large differences in the usage of individual TRGV genes, a pattern conserved across multiple tissues, including thymus, spleen and the gut. A single TRGV gene, TRGV3.3, with a highly diverse private CDR3 repertoire dominated every tissue in all birds. TRGV usage patterns were partly explained by the TRGV-associated recombination signal sequences. Public CDR3 clonotypes represented varying proportions of the repertoire of TCRs utilising different TRGVs, with one TRGV dominated by super-public clones present in all birds. CONCLUSIONS: The application of repertoire analysis enabled functional annotation of the TCRG locus in a species with a high circulating γδ phenotype. This revealed variable usage of TCRGV genes across multiple tissues, a pattern quite different to that found in γδ low species (human and mouse). Defining the repertoire biology of avian γδ T cells will be key to understanding the evolution and functional diversity of these enigmatic lymphocytes in an animal that is numerically more reliant on them. Practically, this will reveal novel ways in which these cells can be exploited to improve health in medical and veterinary contexts.

• MeSH
• genom * MeSH
• genomika MeSH
• kur domácí * genetika   MeSH
• receptory antigenů T-buněk gama-delta * genetika   MeSH
• T-lymfocyty MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články 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

• MeSH
• aktivace lymfocytů MeSH
• buňky kostní dřeně imunologie   patologie   MeSH
• CD antigeny analýza   krev   MeSH
• imunofenotypizace MeSH
• jehlová biopsie MeSH
• lidé MeSH
• lymfoproliferativní nemoci imunologie   krev   patologie   MeSH
• průtoková cytometrie MeSH
• receptory antigenů T-buněk gama-delta * analýza   krev   MeSH
• senioři MeSH
• T-lymfocyty - podskupiny imunologie   patologie   MeSH
• T-lymfocyty * imunologie   patologie   MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• dopisy MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH