Human megakaryocytes. V. Changes in the phenotypic profile of differentiating megakaryocytes

R. B. Levene, J. M D Lamaziere, Hal Broxmeyer, L. Lu, E. M. Rabellino

Research output: Contribution to journalArticle

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Abstract

Human megakaryocytes were studied for phenotypic changes occurring throughout differentiation using a panel of monoclonal antibodies raised against marrow megakaryocytes and blood platelets. 11 monoclonal antibody preparations were selected for restricted specificity against megakaryocytes and/or platelets after screening by immunofluorescence, complement-mediated cytolysis, and solid phase enzyme-linked immunosorbent assay. The expression of the cellular epitopes recognized by these reagents enabled the identification of three levels of megakaryocyte maturation characterized by distinct immunologic phenotypes. Based upon their reactivities against megakarytocyte cells at different ontogenetic levels, monoclonal antibodies were operationally categorized into three groups. Group A consisted of six different monoclonal antibodies that recognized antigens on the colony-forming unit-megakaryocyte (CFU-Mk), in vitro grown colony megakaryocytes, and early immature marrow megakaryocytes, only, and did not detect their respective epitopes on either mature megakaryocytes or platelets. A monoclonal antibody categorized in group B detected a cell antigen expressed by megakaryocytic cells at all maturational levels, but which is lost or suppressed during terminal differentiation and is not expressed on blood platelets. Group C included four different monoclonal antibodies raised against platelets that recognized antigenic determinants expressed on the CFU-Mk, colony megakaryocytes, early and mature megakaryocytes, and platelets. Three group C monoclonal antibodies (PC-1, PC-3, and PC-4) were specific for platelet glycoprotein IIb/IIIa. Additionally, group C monoclonal antibody PC-2 was unique in that it showed partial reactivity against the clonable progenitor for the erythroid series (BFU-E). Recognition of discrete phenotypic changes in differentiating megakaryocytes will enable multiparameter analyses of these cells as well as the study of factors regulating the dynamics of megakaryocytopoiesis in health and disease.

Original languageEnglish (US)
Pages (from-to)457-474
Number of pages18
JournalJournal of Experimental Medicine
Volume161
Issue number3
DOIs
StatePublished - 1985
Externally publishedYes

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Megakaryocytes
Monoclonal Antibodies
Blood Platelets
Epitopes
Stem Cells
Bone Marrow
Thrombopoiesis
Integrin beta3
Antigens
Erythroid Precursor Cells
Platelet Glycoprotein GPIIb-IIIa Complex
Fluorescent Antibody Technique
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Immunology

Cite this

Human megakaryocytes. V. Changes in the phenotypic profile of differentiating megakaryocytes. / Levene, R. B.; Lamaziere, J. M D; Broxmeyer, Hal; Lu, L.; Rabellino, E. M.

In: Journal of Experimental Medicine, Vol. 161, No. 3, 1985, p. 457-474.

Research output: Contribution to journalArticle

Levene, R. B. ; Lamaziere, J. M D ; Broxmeyer, Hal ; Lu, L. ; Rabellino, E. M. / Human megakaryocytes. V. Changes in the phenotypic profile of differentiating megakaryocytes. In: Journal of Experimental Medicine. 1985 ; Vol. 161, No. 3. pp. 457-474.
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