Molecular pathways of early CD105-positive erythroid cells as compared with CD34-positive common precursor cells by flow cytometric cell-sorting and gene expression profiling

Abstract

Special attention has recently been drawn to the molecular network of different genes that are responsible for the development of erythroid cells. The aim of the present study was to establish in detail the immunophenotype of early erythroid cells and to compare the gene expression profile of freshly isolated early erythroid precursors with that of the CD34-positive (CD34+) compartment. Multiparameter flow cytometric analyses of human bone marrow mononuclear cell fractions (n=20) defined three distinct early erythroid stages. The gene expression profile of sorted early erythroid cells was analyzed by Affymetrix array technology. For 4524 genes, a differential regulation was found in CD105-positive erythroid cells as compared with the CD34+ progenitor compartment (2362 upregulated genes). A highly significant difference was observed in the expression level of genes involved in transcription, heme synthesis, iron and mitochondrial metabolism and transforming growth factor-β signaling. A comparison with recently published data showed over 1000 genes that as yet have not been reported to be upregulated in the early erythroid lineage. The gene expression level within distinct pathways could be illustrated directly by applying the Ingenuity software program. The results of gene expression analyses can be seen at the Gene Expression Omnibus repository.

Immunophenotypic analysis of erythroid dysplasia in myelodysplastic syndromes. A report from the IMDSFlow working group

“…Reticulocytes are not covered in these graphs, but they may appear as CD71 dim-to-negative in non-lysed cell preparations. Myeloid progenitors are CD34 + /CD117 + /HLA-DR + /CD105 -( Figure 1C and D.); these cells have slightly higher CD45 expression than erythroid precursors; moreover, in contrast to myeloid progenitors erythroid cells do not express HLA-DR (adapted from references [14][15][16] ).…”

“…The characteristic morphological stages of normal erythroid differentiation are reflected by their light scatter properties and by their differential expression of CD45, CD117, CD105, CD36, CD71 and/or CD235a ( Figure 1). [14][15][16][17] Some of the FC aberrancies that have been reported to reflect MDSrelated dyserythropoiesis are: a) an increased number of NEC within total nucleated cells; b) an altered proportion of consecutive erythroid differentiation stages, such as an increased number of immature erythroid cells (CD117 + and/or CD105 + ) or, by contrast, a decrease in erythroid progenitors; c) an abnormal pattern of CD71 versus CD235a; d) a reduced expression of CD71 and/or CD36; and e) an overexpression of CD105. Most of these aberrancies are present in 70-80% of MDS cases.…”

Prospective isolation of human erythroid lineage-committed progenitors

“…Erythrocytes, like all blood lineages, develop through a series of differentiation stages that begin with HSCs, which have been prospectively isolated in humans (10). The surface markers of intermediate erythroid progenitors/precursors and the expression pattern of some pivotal transcription factors (TFs) during their differentiation from HSCs have been analyzed (11)(12)(13)(14)(15)(16)(17)(18)(19). As a result, the erythroid-committed progenitor (EP), which exists downstream of the bipotent MEP, has been isolated in mouse but not human (9).…”

“…In humans, CD105 and CD71 have been classified as early erythroid cell markers (14)(15)(16)(17)(18)(19). However, CD105 and CD71 expression in oligopotent myeloid progenitors (8) [i.e., CMPs, granulocyte/macrophage progenitors (GMPs), and MEPs] has not been evaluated.…”

Positive p53 immunostaining and erythroid maturation in two cases of pure erythroid leukemia with extremely complex karyotypes

“…With ongoing differentiation, at the second stage, CD36, CD71, CD105, CD117, CD173, and CD238 can be observed, whereas CD45 doi: 10.7243/2052-434X-4-3 expression is diminished and HLA-DR is no longer detectable. The third stage is defined by the appearance of CD235a and the disappearance of CD117 [5,6]. In Case 1, neoplastic cells were positive for CD36, but negative for CD235a, exhibiting the early stage after commitment to the erythroid lineage.…”