Rapid assay of intrinsic radiosensitivity based on apoptosis in human CD4 and CD8 T-lymphocytes

Ozşahin, Mahmut; Özşahin, Hülya; Shi, Yu; Larsson, Birgitta; Würgler, Friedrich E.; Crompton, N. E. A.

doi:10.1016/s0360-3016(97)00038-2

Cited by 107 publications

(66 citation statements)

References 32 publications

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“…It is hoped that an ongoing prospective study will help to define the magnitude of this problem. The reduced apoptosis observed in T-lymphocytes from the hypersensitive individuals supports our hypothesis that such individuals are unable to mount a normal, physiological response to ionizing radiation, and this results in a more severe, pathological reaction (5).…”

Section: Discussionsupporting

confidence: 77%

“…The assay also efficiently demonstrates sensitizing effects of concomitant chemotherapy on the background apoptosis levels in T-lymphocytes. Finally, it is noted that with the increasing number of "dose-escalation" clinical trials, using either conformal radiation therapy or intensity modulated radiation therapy at various sites, the radiation oncologist will welcome a simple and quick normal-tissue radiosensitivity test for use in determining eligibility or stratification criteria of patients to be included in such studies (5). As part of an ongoing prospective clinical trial, these various issues are currently being addressed.…”

Section: Discussionmentioning

confidence: 99%

“…We have previously demonstrated that the population of dead cells evaluated was apoptotic (4,5). The cells display a small size with a reduced propidium iodide signal.…”

Section: Introductionmentioning

confidence: 98%

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Altered apoptotic profiles in irradiated patients with increased toxicity

Ozşahin¹,

Miralbell²,

Emery³

et al. 1998

International Journal of Radiation Oncology*Biology*Physics

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Purpose: A retrospective study of radiation-induced apoptosis in CD4 and CD8 T-lymphocytes, from 12 cancer patients who displayed enhanced toxicity to radiation therapy and 9 ataxia telangiectasia patients, was performed to test for altered response compared to healthy blood-donors and normal cancer patients. Methods and Materials: Three milliliters of heparinized blood from each donor was sent via express post to the Paul Scherrer Institute (PSI) for subsequent examination. The blood was diluted 1:10 in RPMI medium, irradiated with 0-, 2-, or 9-Gy X-rays, and incubated for 48 h. CD4 and CD8 T-lymphocytes were then labeled using FITC-conjugated antibodies, erythrocytes were lysed, and the DNA stained with propidium iodide. Subsequently, cells were analyzed using a Becton Dickinson FACScan flow cytometer. Radiation-induced apoptosis was recognized in leukocytes as reduced DNA content attributed to apoptosis-associated changes in chromatin structure. Apoptosis was confirmed by light microscopy, electron microscopy, and by the use of commercially available apoptosis detection kits (in situ nick translation and Annexin V). Data from hypersensitive individuals were compared to a standard database of 105 healthy blood-donors, and a database of 48 cancer patient blood donors who displayed normal toxicity to radiation therapy. To integrate radiosensitivity results from CD4 and CD8 T-lymphocytes after 2 and 9 Gy, z-score analyses were performed. Results: A cohort of 12 hypersensitive patients was evaluated; 8 showed enhanced early toxicity, 3 showed enhanced late toxicity, and 1 showed both. The cohort displayed less radiation-induced apoptosis (؊1.8 ) than average age-matched donors. A cohort of 9 ataxia telangiectasia homozygotes displayed even less apoptosis (؊3.6 ). Conclusion: The leukocyte apoptosis assay appears to be a useful predictor of individuals likely to display increased toxicity to radiation therapy; however, validation of this requires a prospective study.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

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Altered apoptotic profiles in irradiated patients with increased toxicity

Ozşahin¹,

Miralbell²,

Emery³

et al. 1998

International Journal of Radiation Oncology*Biology*Physics

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“…The data published so far on the cellular and molecular factors underlying acute or late tissue reactions appeared to be contradictory and suggest that there is no clear-cut relationship between cellular radiosensitivity and the risk of acute or late reactions; consequently, no test has been recommended up to now for predicting the risk or the severity of late reactions in breast cancer (Burnet et al, 1992;Brock et al, 1995;Jones et al, 1995;Johansen et al, 1996;Russell et al, 1998;Kiltie et al, 1999;Barber et al, 2000;Peacock et al, 2000;Oppitz et al, 2001). To confirm our first preclinical and retrospective studies on the correlation of radiation-induced CD4 and CD8 T-lymphocyte apoptosis (RTLA) and late side effects after RT (Ozsahin et al, 1997), we assessed prospectively RTLA by the prediction of individual intrinsic radiosensitivity of 399 consenting patients treated with curative RT for miscellaneous cancers (Ozsahin et al, 2003). RTLA significantly predicted grades 2 and 3 late effects (Po0.0001).…”

Section: Discussionmentioning

confidence: 93%

“…Apoptotic lymphocytes were defined as those cells staining positively for their cell-type-specific antibodies, and displaying reduced DNA content and cell size. These lymphocytes were previously examined for apoptotic cells by the TUNEL assay (Ozsahin et al, 1997). Data for at least 10 000 cells sample À1 were acquired.…”

Section: Radiation-induced Apoptosismentioning

confidence: 99%

Concomitant use of tamoxifen with radiotherapy enhances subcutaneous breast fibrosis in hypersensitive patients

et al. 2004

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Concomitant use of adjuvant tamoxifen (TAM) and radiation therapy (RT) is not widely accepted. We aim to assess whether this treatment is associated with an increased risk of developing subcutaneous fibrosis after conservative or radical surgery in breast cancer patients. We analysed 147 women with breast cancer treated with adjuvant RT, and who were included in the KFS 00539-9-1997/SKL 00778-2-1999 prospective study aimed at evaluating the predictive value of CD4 and CD8 T-lymphocyte apoptosis for the development of radiation-induced late effects. TAM (20 mg day À1 ) with concomitant RT was prescribed in 90 hormone receptorpositive patients. There was a statistically significant difference in terms of complication-relapse-free survival (CRFS) rates at 3 years, 48% (95% CI 37.2 -57.6%) vs 66% (95% CI 49.9 -78.6%) and complication-free survival (CFS) rates at 2 years, 51% (95% CI 40 -61%) vs 80% (95% CI 67 -89%) in the TAM and no-TAM groups, respectively. In each of these groups, the CRFS rates were significantly lower for patients with low levels of CD8 radiation-induced apoptosis, 20% (95% CI 10 -31.9%), 66% (95% CI 51.1 -77.6%), and 79% (95% CI 55 -90.9%) for CD8 p16, 16 -24, and 424%, respectively. Similar results were observed for the CFS rates. The concomitant use of TAM with RT is significantly associated with an increased incidence of grade 2 or greater subcutaneous fibrosis; therefore, caution is needed for radiosensitive patients.

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Separation of viable from radiation‐induced apoptotic lymphocytes by free‐flow electrophoresis

et al. 1998

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A human lymphocyte population undergoing apoptosis in vitro due to gamma-irradiation was fractionated by free-flow electrophoresis in triethanolamine--Na-acetate buffers, containing up to 50 mM NaCl, with pH 6.0, 7.2 and 8.5, made isotonic by addition of sucrose. As shown by a flow cytometric analysis of the eluate, the distribution of apoptotic lymphocytes is shifted to the range of higher electrophoretic mobilities relative to that of viable ones at pH 8.5, yielding cell fractions enriched in apoptotic cells by a factor of 3 to 5. The difference in rates of electrophoretic migration observed at a mildly alkaline pH but not at a neutral or mildly acidic one suggests that the surface of apoptotic lymphocytes is more acidic than that of viable ones.

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Rapid assay of intrinsic radiosensitivity based on apoptosis in human CD4 and CD8 T-lymphocytes

Cited by 107 publications

References 32 publications

Altered apoptotic profiles in irradiated patients with increased toxicity

Altered apoptotic profiles in irradiated patients with increased toxicity

Concomitant use of tamoxifen with radiotherapy enhances subcutaneous breast fibrosis in hypersensitive patients

Separation of viable from radiation‐induced apoptotic lymphocytes by free‐flow electrophoresis

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