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Erythropoietin receptor expression on human bone marrow erythroid precursor cells by a newly‐devised quantitative flow‐cytometric assay
Author(s) -
Shinjo Kaori,
Takeshita Akihiro,
Higuchi Masato,
Ohnishi Kazunori,
Ohno Ryuzo
Publication year - 1997
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1997.d01-2071.x
Subject(s) - erythropoietin receptor , cd38 , bone marrow , cd34 , erythropoietin , biology , flow cytometry , microbiology and biotechnology , chemistry , immunology , stem cell , endocrinology
In order to develop a non‐isotopic quantitative assay of erythropoietin (Epo) receptor (EpoR) on human cells, we devised a flow‐cytometric assay using cells stained with biotin‐labelled and a streptavidine–RED670 conjugate. For quantification, we applied the Kolmogorov‐Smirnov test and calculated the D value. The D value was evaluated from the degree of shift in two profiles according to the increase of fluorescence intensity due to the specific binding of biotin‐labelled Epo to EpoR. A good correlation was observed between the number of EpoR calculated by 125 I‐Epo binding assay and the D value. Then, EpoR expression on bone marrow cells from normal individuals was studied by three‐colour flow cytometry. In normal bone marrow, the number of EpoR on cells was highest in CD34 + CD38 − cells (approximately 1600 sites/cell), and decreased in the following order: CD34 + CD38 − cells > CD34 + CD38 + cells > CD34 − CD38 + cells. Glycophorin A (GpA) positive erythroid cells also expressed EpoR, and their CD34 + fraction expressed more EpoR than their CD34 − fraction. However, the expression levels of EpoR of these fractions were lower than CD34 + CD38 − cells. These results indicated that EpoR was highly expressed on CD34 + haemopoietic progenitors from very early stages of differentiation without expression of CD38 antigen, and that the level of expression decreased with erythroid differentiation as well as with various lineage commitment in human bone marrow cells.