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UBC Theses and Dissertations

Isolation of a cell surface antigen specific for human acute myelogenous leukemia cells Shipman, Robert Charles


Methods are described whereby three protein bands (#1, #2 and #3), apparently unique to cells of patients with acute myelogenous leukemia (AML), were isolated on non-reducing poly-acrylamide gels. Antisera raised against all three proteins demonstrated absolute specificity, in the ELISA, for absorbed cell membrane preparations from cells of patients with AML and did not react with comparable cell membrane preparations from cells of normal individuals or those of patients with lympho-proliferative disorders. Inhibition studies showed that AML band #1 (68,000 dalton molecular weight) material was capable of totally inhibiting the reaction, in the ELISA, of all three antisera (anti-AML #1, #2 and #3) with absorbed AML cell membrane preparations. When AML cell membranes were examined in SDS-PAGE, only material identified as AML band #1 was detectable, indicating that AML bands #2 and #3 represented aggregates of AML band #1 or artifacts of non-reducing PAGE and the cell membrane preparative procedures. AML band #1 material was shown to be homogenous by two-dimensional gel electrophoresis with a molecular weight of 68,000 daltons and a mean pI of 7.16. AML band #1 material was also shown to be capable of inhibiting the reaction of anti-AML #1 antiserum with the promyelocytic cell line HL-60, as demonstrated by FACS IV analysis, whereas material from normal cell membranes could not demonstrate this inhibition.

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