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
