ABSTRACT
Erythroid biology research involving rhesus macaques has been applied to several topics including malaria, hemoglobinopathy and gene therapy research. However, analyses of the rhesus red blood cells are limited by the inability to identify and sort those cells in research blood samples using flow cytometry. Here it is reported that the BRIC 6 hybridoma clone raised to the human erythroid surface molecule (referred to as CD233, Band 3, AE1, or SLC4A1) produces cross-reactive and erythroid-specific antibodies for flow cytometric detection and sorting of rhesus macaque erythrocytes.
Subject(s)
Anion Exchange Protein 1, Erythrocyte/immunology , Antibodies, Monoclonal/immunology , Cross Reactions/immunology , Erythrocytes/immunology , Flow Cytometry/methods , Macaca mulatta/immunology , Amino Acid Sequence , Animals , Anion Exchange Protein 1, Erythrocyte/chemistry , Erythrocytes/cytology , Glycophorins/metabolism , Humans , Molecular Sequence DataABSTRACT
Candidate drugs are being sought for the suppression of human erythropoiesis. Cl-IB-MECA [2-chloro-N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide] is a derivative of adenosine that inhibits the growth of leukaemic cell lines. To determine the effects of Cl-IB-MECA upon erythropoiesis, studies were performed by using an ex vivo culture system of primary human CD34+ cells. Cl-IB-MECA suppressed erythroblast growth and maturation at doses >/=50 mumol/l through a mechanism of cell cycle inhibition and accumulation of cells in the G1/G0 phase. These findings demonstrate that Cl-IB-MECA inhibits human erythropoiesis, and suggest that further consideration of this drug is warranted for patients with erythrocytosis or polycythemia syndromes.
Subject(s)
Adenosine/analogs & derivatives , Antineoplastic Agents/pharmacology , Erythropoiesis/drug effects , Adenosine/pharmacology , Antigens, CD34 , Apoptosis/drug effects , Cell Count/methods , Cell Cycle/drug effects , Cells, Cultured , Culture Media , Erythroblasts/drug effects , Erythropoietin , G1 Phase/drug effects , Humans , Polymerase Chain Reaction/methods , Receptor, Adenosine A3/analysis , Resting Phase, Cell Cycle/drug effectsABSTRACT
Here we describe the identification and characterization of an alternate delta-globin mRNA (Alt-d) discovered during high-throughput sequencing of mRNA from adult human erythroid cells. Alt-d mRNA shares the same coding region, splicing pattern, downstream untranslated region, and site of polyadenylation with the previously defined delta-globin (Delta) mRNA. Alt-d mRNA encodes an additional 145 nt in the upstream untranslated region, suggesting an alternative site of transcriptional initiation and transcription through the previously defined promoter, which contains several protein-binding motifs and a TATA box. Northern blot and PCR analyses demonstrated a restricted expression of Alt-d in fetal liver, bone marrow, and adult reticulocytes. Quantitative PCR demonstrated an Alt-d expression pattern similar to that of the Delta transcripts. In addition to intergenic RNA species and the dominant delta-globin transcripts, these data suggest that a third form of RNA is produced from low-level transcription through the delta-globin gene promoter.
