Constitutive expression of p53 protein in enriched normal human marrow blast cell populations.
Previous studies by others using metabolic labeling, cell lysis, and immunoprecipitation have reported elevated levels of p53 protein in blast cells derived from patients with acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML), whereas p53 protein was not detected in normal light-density bone marrow cells. In this report, using the same detection methods, we confirm the negligible expression of p53 protein in normal light density marrow cells. However, we find clearly significant levels of p53 protein expression in enriched normal human marrow blast populations. Furthermore, using a panel of p53 specific monoclonal antibodies, we find the p53 protein constitutively synthesized by normal marrow blasts has the immunologic phenotype identified by PAb240 that reportedly recognizes a common conformational-dependent epitope on mutant p53. We have also found that the p53 immunologic subclass identified by PAb240 exists in normal human circulating lymphocytes either resting, serum starved, or PHA activated. In summary, it is clear that (1) normal marrow blast populations provide the appropriate control for assessing the levels of p53 protein expression in leukemic blast cells; and (2) PAb240 cannot be used to distinguish p53 mutated at the DNA level from normal p53 in fresh human hematopoietic cells.
Rivas
C I
CI
Memorial Sloan-Kettering Cancer Center, Laboratory of Hematopoietic Cell Kinetics, New York, NY 10021.
Wisniewski
D
D
Strife
A
A
Perez
A
A
Lambek
C
C
Bruno
S
S
Darzynkiewicz
Z
Z
Clarkson
B
B
D000911:Antibodies, Monoclonal; D001345:Autoradiography; D001853:Bone Marrow / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D006801:Humans; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000502:physiology*; D008715:Methionine / Q000378:metabolism; D012016:Reference Values; D013462:Sulfur Radioisotopes; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D013462:Sulfur Radioisotopes; D016159:Tumor Suppressor Protein p53; D008715:Methionine