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Monoclonal antibody YB5.B8 identifies the human c-kit protein product
NB Lerner, KH Nocka, SR Cole, FH Qiu, A Strife, LK Ashman and P Besmer
Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York,
NY.
The c-kit proto-oncogene encodes a 145- to 160-Kd transmembrane tyrosine
kinase, which is a member of the platelet-derived growth factor receptor
family and is allelic with the murine white spotting locus (W). W mutations
affect several aspects of hematopoiesis, most notably erythroid progenitors
and mast cells. A monoclonal antibody, YB5.B8, had been raised against the
leukemic blasts of a patient with M1-type acute myelocytic leukemia (AML)
and it precipitates a 150-Kd cell surface glycoprotein from leukemic cells.
The YB5.B8 epitope is expressed on mast cells, on up to 3% of normal
mononuclear bone marrow cells, and it identifies a sub-group of AML
patients with a poor prognosis. In view of similarities noted between the
cell surface antigen identified by YB5.B8 and the c-kit protein product, we
performed experiments to determine whether they are identical. c-kit RNA
expression in the cell lines HEL (human erythroleukemia) and A172
(glioblastoma) was shown to parallel the expression of the YB5.B8 epitope
in these lines as measured by flow cytometry. Immunoprecipitation analysis
with anti-kit serum and YB5.B8 antibody indicated that the two antibodies
identified proteins of identical size in HEL (155 Kd) and A172 (145 Kd)
cells, and sequential immunoprecipitations with the kit and the YB5.B8
antibodies demonstrated that the two antibodies recognize the same
molecule. The proteins identified by both the anti-kit and YB5.B8
antibodies displayed in vitro autophosphorylation activity in immune
complex kinase assays. In addition, YB5.B8 was able to inhibit the binding
of the kit ligand to HEL cells. These studies provide evidence that the
YB5.B8 antigen and the c-kit protein product are identical and raise
certain hypotheses regarding the role of c-kit in AML.
Volume 77,
Issue 9,
pp. 1876-1883,
05/01/1991
Copyright © 1991 by The American Society of Hematology
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