The CD8+ natural killer (NK) subpopulation has recently been identified as a fast and reliable biodosimetric indicator within human peripheral blood mononuclear cells (PBMC) in vitro. In irradiated and subsequently cultivated PBMC, a decrease of the relative number of intact CD3-CD8+ lymphocytes 16 and 48 h after treatment has allowed for estimating the received dose in the range of 0 - 10 Gy and lethal/sublethal dose discrimination, respectively. Here we show that suitable biodosimeters can also be found in the peripheral blood B-cell compartment. Multiparameter flow cytometric analysis of irradiated and subsequently cultivated human PBMC revealed that both the CD27+ and CD21- B-cell subpopulations can be used as biodosimeters and the CD19+CD27+ lymphocytes have proved useful for retrospective determination of the received dose in the range of 0 - 6 Gy. In addition, several CD19+ lymphocyte subsets characterized by co-expression of CD21, CD27 and CD38 have been shown to bear biodosimetric potential, too. However, when important parameters like the original size within the CD19+ compartment, its radiation-induced changes and data variation had been taken into account, the CD27+ subpopulation proved superior to the other B-cell subpopulations and subsets. It appears that, in the dose range of 0 - 6 Gy, the relative decrease of CD27+ B lymphocytes provides more sensitive and reliable data than that of CD8+ NK-cells due mainly to lower data variation. In contrast to CD27+ B-cells, the proportions of CD27+ subpopulations of T-cells were not affected by irradiation. We have also proposed a simple experimental protocol based on full blood cultivation and three-color CD27/CD3/CD19 immunophenotyping as a time-saving and inexpensive approach for practical biodosimetric evaluations on simple, three-to-four color flow cytometers.

• Klíčová slova
• B-cells, CD27, Biodosimetric marker, ?-irradiation,
• MeSH
• annexin A5 metabolismus   MeSH
• antigeny CD27 fyziologie   MeSH
• antigeny CD38 fyziologie   MeSH
• B-lymfocyty fyziologie   účinky záření   MeSH
• biologické markery MeSH
• fenotyp MeSH
• financování organizované MeSH
• fosfatidylseriny metabolismus   MeSH
• lidé MeSH
• monocyty fyziologie   MeSH
• podskupiny lymfocytů fyziologie   MeSH
• průtoková cytometrie MeSH
• receptory buněčného povrchu metabolismus   MeSH
• receptory komplementu 3d metabolismus   MeSH
• separace buněk MeSH
• vztah dávky záření a odpovědi MeSH
• záření gama MeSH
• Check Tag
• lidé MeSH

The expression of CD27 and CD44 correlate with the genotype of B-precursor acute lymphoblastic leukemia (ALL). Based on the expression of these antigens, we identified counterparts of TEL/AML1(pos) and TEL/AML1(neg) leukemic cells in nonmalignant bone marrow. Although CD27 is known as a marker of mature memory B cells, we recently showed that CD27 is also expressed by malignant and nonmalignant B precursors. Here, we show that CD27 and CD44 delineate stages of B-precursor development. Well-established differentiation markers showed that the developmental sequence starts from undetermined progenitors, expressing CD44. Upon B-lineage commitment, cells gain CD27 and lose CD44. The CD27(pos)CD44(neg) (CD27 single positive, 27SP) cells are the earliest stage within CD10(pos)CD19(pos) B precursors and express RAG-1 and TDT. These cells correspond to TEL/AML1(pos) ALL (1/4 pediatric B-precursor ALL). The development follows to CD27/CD44 double-positive (27/44DP) stage, 44SP stage and CD27/CD44 double-negative (27/44DN) stage. Before exit to periphery, CD44 is reexpressed. The 27/44DP cells are mostly large and profoundly suppress RAG-1. Despite their presumably high proliferation potential, 27/44DP cells rarely dominate in leukemia. At 44SP stage, which corresponds to TEL/AML1(neg) leukemias, RAG-1 is reexpressed and Ig light chain gene starts to be rearranged.

• MeSH
• antigeny CD27 biosyntéza   fyziologie   genetika   MeSH
• antigeny CD44 biosyntéza   fyziologie   genetika   MeSH
• dítě MeSH
• financování organizované MeSH
• genová přestavba B-lymfocytů imunologie   MeSH
• imunofenotypizace MeSH
• leukemie B-buněčná diagnóza   genetika   imunologie   MeSH
• lidé MeSH
• lymfopoéza genetika   imunologie   MeSH
• prekurzorové B-lymfoidní buňky cytologie   imunologie   patologie   MeSH
• vývojová regulace genové exprese MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• srovnávací studie MeSH

Advances in single-cell level analytical techniques, especially cytometric approaches, have led to profound innovation in biomedical research, particularly in the field of clinical immunology. This has resulted in an expansion of high-dimensional data, posing great challenges for comprehensive and unbiased analysis. Conventional manual analysis is thus becoming untenable to handle these challenges. Furthermore, most newly developed computational methods lack flexibility and interoperability, hampering their accessibility and usability. Here, we adapted Seurat, an R package originally developed for single-cell RNA sequencing (scRNA-seq) analysis, for high-dimensional flow cytometric data analysis. Based on a 20-marker antibody panel and analyses of T-cell profiles in both adult blood and cord blood (CB), we showcased the robust capacity of Seurat in flow cytometric data analysis, which was further validated by Spectre, another high-dimensional cytometric data analysis package, and conventional manual analysis. Importantly, we identified a unique CD8+ T-cell population defined as CD8+CD45RA+CD27+CD161+ T cell that was predominantly present in CB. We characterised its IFN-γ-producing and potential cytotoxic properties using flow cytometry experiments and scRNA-seq analysis from a published dataset. Collectively, we identified a unique human CB CD8+CD45RA+CD27+CD161+ T-cell subset and demonstrated that Seurat, a widely used package for scRNA-seq analysis, possesses great potential to be repurposed for cytometric data analysis. This facilitates an unbiased and thorough interpretation of complicated high-dimensional data using a single analytical pipeline and opens a novel avenue for data-driven investigation in clinical immunology.

• MeSH
• analýza jednotlivých buněk metody   MeSH
• antigeny CD27 metabolismus   imunologie   MeSH
• antigeny CD45 * metabolismus   imunologie   MeSH
• CD8-pozitivní T-lymfocyty * imunologie   MeSH
• dospělí MeSH
• fetální krev * imunologie   cytologie   MeSH
• imunofenotypizace metody   MeSH
• lektinové receptory NK-buněk - podrodina B imunologie   metabolismus   MeSH
• lidé MeSH
• průtoková cytometrie * metody   MeSH
• software MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• akutní lymfatická leukemie diagnóza   genetika   imunologie   MeSH
• antigeny CD27 analýza   biosyntéza   genetika   MeSH
• antigeny CD44 analýza   biosyntéza   genetika   MeSH
• genotyp MeSH
• imunofenotypizace MeSH
• kohortové studie MeSH
• lidé MeSH
• prognóza MeSH
• průtoková cytometrie * metody   MeSH
• stanovení celkové genové exprese * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

We have shown previously that ETV6/RUNX1-positive acute lymphoblastic leukemia (ALL) is distinguishable from other ALL subtypes by CD27pos /CD44low-neg immunophenotype. During diagnostic immunophenotyping of 573 childhood B-cell precursor ALL (BCP-ALL), we identified eight cases with this immunophenotype among "B-other ALL" (BCP-ALL cases negative for routinely tested chromosomal/genetic aberrations). We aimed to elucidate whether these cases belong to the recently described ETV6/RUNX1-like ALL defined by the ETV6/RUNX1-specific gene expression profile (GEP), harboring concurrent ETV6 and IKZF1 lesions. We performed comprehensive genomic analysis using single nucleotide polymorphism arrays, whole exome and transcriptome sequencing and GEP on microarrays. In unsupervised hierarchical clustering based on GEP, five out of seven analyzed CD27pos /CD44low-neg B-other cases clustered with ETV6/RUNX1-positive ALL and were thus classified as ETV6/RUNX1-like ALL. The two cases clustering outside ETV6/RUNX1-positive ALL harbored a P2RY8/CRLF2 fusion with activating JAK2 mutations and a TCF3/ZNF384 fusion, respectively, assigning them to other ALL subtypes. All five ETV6/RUNX1-like cases harbored ETV6 deletions; uniform intragenic ARPP21 deletions and various IKZF1 lesions were each found in three ETV6/RUNX1-like cases. The frequency of ETV6 and ARPP21 deletions was significantly higher in ETV6/RUNX1-like ALL compared with a reference cohort of 42 B-other ALL. In conclusion, we show that ETV6/RUNX1-like ALL is associated with CD27pos /CD44low-neg immunophenotype and identify ARPP21 deletions to contribute to its specific genomic profile enriched for ETV6 and IKZF1 lesions. In conjunction with previously published data, our study identifies the ETV6 lesion as the only common genetic aberration and thus the most likely key driver of ETV6/RUNX1-like ALL.

• MeSH
• akutní lymfatická leukemie krev   genetika   imunologie   MeSH
• antigeny CD27 genetika   metabolismus   MeSH
• antigeny CD44 genetika   metabolismus   MeSH
• B-lymfocyty imunologie   MeSH
• dítě MeSH
• fenotyp * MeSH
• fúzní onkogenní proteiny genetika   MeSH
• imunofenotypizace MeSH
• jednonukleotidový polymorfismus MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• protein PEBP2A2 genetika   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Publikační typ
• abstrakty MeSH

• MeSH
• analýza přežití MeSH
• antigeny CD27 účinky léků   MeSH
• lidé MeSH
• melanom * farmakoterapie   imunologie   MeSH
• metastázy nádorů * farmakoterapie   imunologie   MeSH
• monoklonální protilátky farmakologie   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• klinické zkoušky, fáze III MeSH
• novinové články MeSH

B cells play an important role in the immune responses which affect the outcomes of kidney allografts. Dynamic changes of B-cell compartments in clinical kidney transplantation are still poorly understood. B-cell subsets were prospectively monitored using flow cytometry for 1 year in 98 kidney transplant recipients. Data were correlated with immunosuppression and clinical outcomes. An increase in the total population of B lymphocytes was observed during the first week after transplantation. The level of IgM(high) CD38(high) CD24(high) transitional B cells reduced significantly up until the third month, with partial repopulation in the first year. Lower numbers of transitional B cells in the third month were associated with higher risk of graft rejection. IgM(+) IgD(+) CD27(-) naive B cells did not change within follow-up. IgM(+) CD27(+) nonswitched memory B cells and IgM(-) CD27(+) switched memory B cells increased on post-operative day 7. IgM(-) CD38(high) CD27(high) plasmablasts showed similar kinetics during the first post-transplant year, similar to transitional B cells. In conclusion, sensitized kidney transplant recipients as well as those with either acute or chronic rejection within the first post-transplant year exhibited lower levels of transitional B cells. Therefore, these data further support the hypothesis that transitional B cells have a protective role in kidney transplantation.

• MeSH
• alografty MeSH
• antigen CD24 metabolismus   MeSH
• antigeny CD27 metabolismus   MeSH
• antigeny CD38 metabolismus   MeSH
• časové faktory MeSH
• dítě MeSH
• dospělí MeSH
• imunologická paměť imunologie   MeSH
• imunosupresivní léčba MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• plazmatické buňky imunologie   MeSH
• podskupiny B-lymfocytů imunologie   MeSH
• předškolní dítě MeSH
• prekurzorové B-lymfoidní buňky imunologie   MeSH
• příjemce transplantátu MeSH
• prospektivní studie MeSH
• rejekce štěpu imunologie   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• transplantace ledvin * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• antigeny CD27 * krev   MeSH
• antigeny sacharidové asociované s nádorem * krev   MeSH
• erytrocyty * imunologie   MeSH
• isoantigeny * krev   MeSH
• koně imunologie   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• monoklonální protilátky MeSH
• reprodukovatelnost výsledků MeSH
• sacharidové sekvence MeSH
• zkřížené reakce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• komentáře MeSH

Chronic kidney disease stage 5 (CKD5) dialysis patients who stay long term in uremic environment often exhibit several, poorly defined, immune impairments. In this study, we assessed peripheral virus-specific effector/memory cells and subpopulations of T, B and DC cells using ELISPOT and FACS methods in 74 low-risk kidney transplant candidates without anti-HLA antibodies, prior to transplantation in pre-emptive (never experienced dialysis) and dialysis cohorts. There was difference in circulating marginal zone B cells (MZB) (IgDhighCD27high) between dialysis patients and those receiving kidney grafts pre-emptively (P = .002). Patients treated on dialysis >12 months had also 4.2-fold greater risk of increased absolute numbers of MZB (95%CI:1.6-11.2; P = .004). There were no other differences in B-, T- and DC-cell subsets. Numbers of effector/memory T cells reactive to major opportunistic virus-specific antigens (CMV, BKV and EBV) were not affected by dialysis. Non-sensitised dialysis-treated patients displayed significantly more circulating MZB compared to those CKD5 patients that had never undergone dialysis therapy.

• MeSH
• antigeny CD27 metabolismus   MeSH
• B-lymfocyty imunologie   MeSH
• chronická renální insuficience imunologie   terapie   MeSH
• dendritické buňky imunologie   MeSH
• dialýza MeSH
• dospělí MeSH
• ELISPOT MeSH
• imunologická paměť MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• senioři MeSH
• slezina patologie   MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH