CD27(+) peripheral blood B-cells are a useful biodosimetric marker in vitro
Z Řeháková1, J Sinkora, M Vlková
1Department of Radiobiology, Faculty of Military Health Sciences, Hradec Králové, Czech Republic.
Insights
New research identifies CD27(+) B lymphocytes as a superior biodosimeter for radiation dose estimation. This method offers a more sensitive and reliable alternative to CD8(+) NK-cells for assessing radiation exposure in peripheral blood.
Area of Science:
- Immunology and Radiation Biology
- Biodosimetry and Radiation Effects
Background:
- CD8(+) natural killer (NK) cells are established biodosimetric indicators in peripheral blood mononuclear cells (PBMC) for estimating radiation dose.
- Previous studies utilized the decrease in CD8(+)CD3(-) lymphocytes post-irradiation for dose assessment (0-10 Gy).
Purpose of the Study:
- To investigate B-cell subpopulations in peripheral blood as potential biodosimeters for radiation exposure.
- To compare the efficacy of B-cell subpopulations against CD8(+) NK-cells for biodosimetry.
Main Methods:
- Multiparameter flow cytometry was used to analyze irradiated human PBMC.
- Evaluated CD27(+) and CD21(-) B-cell subpopulations, along with CD19(+) lymphocyte subsets expressing CD21, CD27, and CD38.
- Developed a simplified protocol using whole blood culture and three-color immunophenotyping (CD27/CD3/CD19).
Main Results:
- CD27(+) and CD21(-) B-cell subpopulations demonstrated biodosimetric potential.
- CD19(+)CD27(+) lymphocytes accurately determined doses between 0-6 Gy.
- The CD27(+) B-cell subpopulation showed superior sensitivity and reliability compared to CD8(+) NK-cells, with lower data variation in the 0-6 Gy range.
Conclusions:
- CD27(+) B lymphocytes are effective and reliable biodosimeters for retrospective radiation dose assessment.
- The proposed whole blood culture and immunophenotyping protocol offers a time-saving and cost-effective approach for practical biodosimetry.
- B-cell subpopulations, particularly CD27(+) B cells, present a promising alternative to NK-cells for radiation biodosimetry.
Abstract:
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 immuno-phenotyping as a time-saving and inexpensive approach for practical biodosimetric evaluations on simple, three-to-four color flow cytometers.
