It has been estimated that 1 to 3% of the general population may be ataxia-telangiectasia (A-T) heterozygotes and hypersensitive to conventional doses of radiation. We attempted to identify heterozygotes by evaluating the proportion of cells in various phases of the cell cycle in response to irradiation. This was accomplished by using flow cytometry to study lymphoblastoid cell lines (LCLs) from 14 A-T homozygotes, 17 genotypic A-T heterozygotes, and 18 normal individuals, including 10 genotypic normals. The LCLs were exposed to 2-gRay radiation and were analyzed after 24 hr along with nonirradiated controls. The difference between the percentage of cells in G2/M with and without irradiation after 24 hr ranged, respectively, from: 12.0 to 31.5% (mean = 18.7 +/- 5.5) for A-T homozygotes; 6.7 to 19.3% (mean = 12.5 +/- 3.8) for A-T heterozygotes; and -1.5 to 12.4% (mean = 6.4 +/- 3.2) for normals. A cut-off region of 9.6 to 13.2% defined by one standard deviation above the mean for normals and one standard deviation below the mean for A-T homozygotes served as the grey zone between normals and A-T heterozygotes or homozygotes. Two of the 18 normals overlapped with the grey zone. Four of 17 heterozygotes were within the normal range; seven fell within the grey zone. This may reflect nongenetic variables, such as the status of the LCLs at the time of testing. Flow cytometry cell-cycle analysis on irradiated LCLs is a useful adjunctive test for establishing a diagnosis of A-T in questionable cases.(ABSTRACT TRUNCATED AT 250 WORDS)