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Rapid Mixed Leukocyte Test with 3H-Acridine Orange1.
CALIFORNIA UNIV SAN FRANCISCO MEDICAL CENTER
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The report concerns search for a tritium labeled compound which would maximally label lymphocytes in mixed culture MLC within 2 - 24 hours. For this purpose tritiated acridine orange 3H-AO was tested. Interest in acridine orange AO stemmed from the report of J. Andersson et al, that by cytophotofluorometric methods, an increase in fluorescence intensity of fixed and AO-stained lymphocytes can be observed 1-3 hours after human allogeneic lymphocytes have been committed to mixed culture. No such increase occurred in lymphocytes from donors compatible at the major histocompatibility locus HL-A. Data confirms that of Andersson et al., but by a method using liquid scintillation spectrophotometric measurement of 3H-acridine orange binding which is more available than estimation of bound acridine orange by spectrofluorometric fluorescence. Also, a method is described for the preparation of a pure tritiated acridine orange of high specific radioactivity.
APPROVED FOR PUBLIC RELEASE