Detection of GD3 ganglioside in childhood acute lymphoblastic leukemia with monoclonal antibody to GD3: Restriction to immunophenotypically defined T-cell disease

Authors

  • William D. Merritt,

    1. Bacterial Toxins Branch, Division of Bacterial Products, Center for Drugs and Biologics, Food and Drug Administration, Bethesda, Maryland 20892
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  • Marcelo B. Sztein,

    1. Department of Medicine, The George Washington University School of Medicine and Health Sciences, Washington, D.C. 20037
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  • Gregory H. Reaman

    1. Department of Child Health and Development, The George Washington University School of Medicine and Health Sciences, Washington, D.C. 20037
    2. The Children's Hospital National Medical Center, Washington, D.C. 20010
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Abstract

We have recently reported that the disialoganglioside GD3 is found in cellular lipid extracts of T-cell acute lymphoblastic malignancies (T-ALL) but is not detectable by resorcinol staining in extracts of non-T acute lymphoblastic leukemia blasts (non-T-ALL). We have now extended this study to assess the detectability of GD3 in T-ALL vs non-T-ALL utilizing an anti-GD3 antibody, R24. Gangliosides isolated from T-ALL and non-T-ALL blasts by two different methods were separated by thin-layer chromatography and stained with anti-GD3 and a control antibody specific for GM3 and sialosylparagloboside (SPG). Anti-GD3 reactivity was observed in extracts from T-ALL cells in all cases, whereas GD3 was not detected in any of the non-T-ALL samples. The anti-GM3/SPG antibody stained GM3 in all of the leukemic samples analyzed as well as SPG in the non-T-ALL samples. Indirect immunofluorescence was used to assess the expression of GD3 at the surface of leukemic blasts. Fluorescence-activated cell sorting analysis with R24 showed that whereas T-ALL blasts were highly reactive with this antibody, non-T-ALL blasts were totally unreactive. In an analysis of a larger number of leukemia patients by fluorescence microscopy, 20 out of 28 samples with the T-ALL phenotype were positive for R24, whereas zero out of 11 non-T-ALL samples were reactive. These results confirm our earlier finding of the specificity of GD3 to the T-ALL subclass of childhood leukemias and furthermore suggest the potential value of anti-GD3 as an immunological tool for the diagnosis and therapy of T-cell ALL.

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