Cellular regulation of i and I antigen expressions in human erythroblasts grown in vitro.

In order to study the cellular regulation of i and I antigen expressions n erythroid cells, burst-forming-unit erythroids were grown in methylcellulose from the blood of normal human adults. Erythroid colonies were harvested on days 9, 11 or 14 of culture. Individual colonies or pooled colonies were subsequently cytocentrifuged and then i or I antigens were studied by immunofluorescence using a monoclonal anti-i or I antibody. Labelled cells were firstly located, their erythroid origin and their stage of maturation were subsequently ascertained after May-Grünwald-Giemsa staining. This approach shows that: (1) a marked heterogeneity occurred from one colony to another, most of the colonies being i-negative while the others contained a mixed population composed by a minority of i-positive erythroid cells; (2) the proportion of both i colonies and i cells decreased with the aging of culture; (3) i antigen was preferentially expressed on immature erythroblasts; however, immaturity per se was not sufficient to determine i expression; (4) I antigen expression was also related to the process of erythroid maturation but in an inverse way; its expression was higher in mature erythroblasts than in immature ones.