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DIAGNOSIS OF LESCH-NYHAN HETEROZYGOTES USING PERIPHERAL BLOOD: 97

Although carriers of deficient genes for Lesch-Nyhan syndrome are detectable using cultured fibroblasts or hair roots, peripheral blood has been insufficient specimen for the diagnosis. We have established a method using peripheral T-cells for the diagnosis of the heterozygotes. Mononuclear cells from five mothers of Lesch-Nyhan boys were stimulated with phytohemagglutinin and then maintained in the medium containing T-cell growth factor. The growing viable T-cells were then exposed to 6-thioguanine. Most of the cells were killed by the treatment, but after 2-4 days, rapid growth of 6-thioguanine resistant cells was observed. T-cells from Lesch-Nyhan boys were totally resistant to 6-thioguanine and normal T-cells were sensitive. We estimated the percentages of HGPRT negative cells among T-cell growth factor-responsive T-cells from each heterozygote by comparing the growth curves for these cell populations with those for mixed cell populations containing enzyme-negative (Lesch-Nyhan's cells) cells and positive (normal) cells in various ratios. The Lesch-Nyhan mothers were estimated to contain 5-40% enzyme negative T-cells, indicating that mosaicism is present in T-cell populations in heterozygotes and the carrier state for Lesch-Nyhan syndrome is detectable using peripheral blood.