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Carbonic anhydrase I is an early specific marker of normal human erythroid
differentiation
JL Villeval, U Testa, G Vinci, H Tonthat, A Bettaieb, M Titeux, P Cramer, L Edelman, H Rochant and J Breton-Gorius
The expression of carbonic anhydrase (CA) as a marker of erythroid
differentiation was investigated by immunologic and enzymatic procedures. A
polyclonal anti-CA antibody was obtained by immunizing rabbits with
purified CA I isozyme. This antibody is reactive with CA I but not with CA
II. Within blood cells, CA I was only present in erythrocytes, whereas CA
II was also detected in platelet lysates by enzymatic assay. Concerning
marrow cells, identifiable erythroblasts and some blast cells expressed CA
I. Most of the glycophorin A-positive marrow cells were clearly labeled by
the anti-CA I antibody. However, rare CA I-positive cells were not reactive
with anti-glycophorin A antibodies. We therefore investigated whether these
cells were erythroid precursors or progenitors. In cell sorting experiments
of marrow cells with the FA6 152 monoclonal antibody, which among
hematopoietic progenitors is reactive only with CFU-E and a part of BFU- E,
was performed, CA I+ cells were found mainly in the positive fraction. The
percentage of CA I+ cells nonreactive with anti- glycophorin A antibodies
contained in the two fractions was in the same range as the percentage of
erythroid progenitors identified by their capacity to form colonies. In
addition, the anti-CA I antibody labeled blood BFU-E-derived colonies as
early as day 6 of culture, whereas in similar experiments with the
anti-glycophorin A antibodies, they were stained three or four days later.
No labeling was observed in CFU-GM- or CFU-MK-derived colonies. The
phenotype of the day 6 cells expressing CA I was similar to that of
erythroid progenitors (CFU-E or BFU-E): negative for glycophorin A and
hemoglobin, and positive for HLA-DR antigen, the antigen identified by FA6
152, and blood group A antigen. Among the cell lines tested, only HEL cells
expressed CA I, while K562 was unlabeled by the anti-CA I antibody. In
contrast, HEL and K562 cells expressed CA II as detected by a biochemical
technique. Synthesis of CA I, as with other erythroid markers such as
glycophorin A and hemoglobin, was almost abolished after
12-O-tetradecanoyl-phorbol-13 acetate treatment of HEL cells. In
conclusion, CA I appears to be an early specific marker of the erythroid
differentiation, expressed by a cell with a similar phenotype as an
erythroid progenitor.
Volume 66,
Issue 5,
pp. 1162-1170,
11/01/1985
Copyright © 1985 by The American Society of Hematology
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