Individual radiosensitivity does not correlate with radiation-induced apoptosis in lymphoblastoid cell lines or CD3+ lymphocytes.
OBJECTIVE
Spontaneous and radiation-induced apoptosis of lymphoblastoid cell lines (LCLs) derived from healthy donors, cancer patients and donors with radiosensitivity syndromes as well as CD(3+) lymphocytes from patients with > or = grade 3 late toxicity were investigated as a possible marker for the detection of individual radiosensitivity. These investigations are based on the hypothesis that hypersensitive patients have reduced levels of apoptosis after in vitro irradiation as a result of a defect in the signaling pathway.


METHODS
Epstein-Barr virus-(EBV-)transformed LCLs derived from five healthy donors, seven patients with heterozygous or homozygous genotype for ataxia-telangiectasia or Nijmegen breakage syndrome and five patients with > or = grade 3 late toxicity (RTOG) were investigated. In addition, CD(3+) lymphocytes from 21 healthy individuals and 18 cancer patients including five patients with a proven cellular hypersensitivity to radiation were analyzed. Cells were irradiated in vitro with a dose of 2 and 5 Gy and were incubated for 48 h. Apoptotic rates were measured by the TUNEL assay followed by customized image analysis.


RESULTS
Four out of seven radiosensitivity syndrome patients were identified to have an increased cellular radiosensitivity as determined by reduced apoptotic rates after irradiation of their respective LCLs. Comparatively, only two of the five hypersensitive cancer patients were clearly identified by reduced apoptotic rates. Spontaneous apoptotic rates were very homogeneous among all 39 samples from controls and patients, while lymphocytes of all cancer patients showed significantly lower radiation-induced rates.


CONCLUSIONS
Only a subgroup of hypersensitive patients may be identified by reduction of radiation-induced apoptotic rate. It is concluded that the hypothesis according to which hypersensitive cells have reduced levels of apoptosis is only conditionally true. The authors suggest that this assay can be used in combination with additional tests evaluating DNA double-strand break repair, cell-cycle control and chromosomal aberrations for the evaluation for individual hypersensitivity.
Wistop
Anja
A
Department of Radiation Oncology, Friedrich Alexander University Erlangen-Nuremberg, Erlangen, Germany.
Keller
Ulrike
U
Sprung
Carl N
CN
Grabenbauer
Gerhard G
GG
Sauer
Rolf
R
Distel
Luitpold V R
LV
D017209:Apoptosis / Q000528:radiation effects*; D001260:Ataxia Telangiectasia / Q000473:pathology; D001782:Blood Donors; D017252:CD3 Complex / Q000032:analysis; D002460:Cell Line; D002461:Cell Line, Transformed; D004854:Herpesvirus 4, Human; D006801:Humans; D020287:In Situ Nick-End Labeling; D011836:Radiation Tolerance; D012016:Reference Values; D013601:T-Lymphocytes / Q000276:immunology / Q000528:radiation effects*; D014407:Tumor Cells, Cultured / Q000528:radiation effects*
D017252:CD3 Complex