The fucosylated histo-blood group antigens H type 2 (blood group O, CD173) and Lewis Y (CD174) are expressed on CD34+ hematopoietic progenitors but absent on mature lymphocytes

Glycobiology, Aug 2001

The expression of LeY, H2, H3, and H4 on a broad variety of human leukemia cell lines and native lymphocytes as well as on CD34+ hematopoietic progenitor cells was examined by flow cytometry and immunocytochemistry. CD34+ leukemia cell lines (KG1, KG1a, and TF1) and native CD34+ hematopoietic progenitor cells expressed H2 (CD173) and LeY (CD174). In contrast, CD34– cell lines (HL-60, U937, JOK-1, Raji, Molt-3, Jurkat, and CEM-C7) and mature lymphocytes from peripheral blood and tonsils lacked CD173 and CD174. All cell lines and native lymphocytes as well as CD34+ precursor cells were negative for H3 and H4. Immunoprecipitation and consecutive Western blotting revealed a 170-kDa glycoprotein as the carrier molecule for the CD173 and CD174 oligosaccharide sequences on CD34+ hematopoietic precursors. The key enzyme for generating CD173 is the β-D-galactoside 2-α-L-fucosyltransferase (FUT1). As shown by RT-PCR, FUT1 was expressed in immature hematopoietic cells but absent in mature lymphocytes, which indicates that expression of CD173 within the hematopoietic system is regulated at the transcriptional level by FUT1. Due to their exclusive presence on CD34+ hematopoietic progenitor cells, CD173 and CD174 represent novel markers of early hematopoiesis. The expression of the fucosylated histo-blood group antigens CD173 and CD174 in CD34+ hematopoietic progenitor cells and down-regulation of FUT1 in mature lymphocytes may be important factors influencing the homing process of hematopoietic stem cells to the bone marrow.

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The fucosylated histo-blood group antigens H type 2 (blood group O, CD173) and Lewis Y (CD174) are expressed on CD34+ hematopoietic progenitors but absent on mature lymphocytes

Yi Cao 1 Anette Merling 1 Uwe Karsten 0 Reinhard Schwartz-Albiez 1 0 Max Delbruck Centre for Molecular Medicine , 13125 Berlin-Buch , Germany 1 Division of Cellular Immunology, German Cancer Research Centre , Im Neuenheimer Feld 280, 69120 Heidelberg , Germany - The expression of LeY, H2, H3, and H4 on a broad variety of human leukemia cell lines and native lymphocytes as well as on CD34+ hematopoietic progenitor cells was examined by flow cytometry and immunocytochemistry. CD34+ leukemia cell lines (KG1, KG1a, and TF1) and native CD34+ hematopoietic progenitor cells expressed H2 (CD173) and LeY (CD174). In contrast, CD34 cell lines (HL-60, U937, JOK-1, Raji, Molt-3, Jurkat, and CEM-C7) and mature lymphocytes from peripheral blood and tonsils lacked CD173 and CD174. All cell lines and native lymphocytes as well as CD34+ precursor cells were negative for H3 and H4. Immunoprecipitation and consecutive Western blotting revealed a 170-kDa glycoprotein as the carrier molecule for the CD173 and CD174 oligosaccharide sequences on CD34+ hematopoietic precursors. The key enzyme for generating CD173 is the -D-galactoside 2--L-fucosyltransferase (FUT1). As shown by RT-PCR, FUT1 was expressed in immature hematopoietic cells but absent in mature lymphocytes, which indicates that expression of CD173 within the hematopoietic system is regulated at the transcriptional level by FUT1. Due to their exclusive presence on CD34+ hematopoietic progenitor cells, CD173 and CD174 represent novel markers of early hematopoiesis. The expression of the fucosylated histo-blood group antigens CD173 and CD174 in CD34+ hematopoietic progenitor cells and down-regulation of FUT1 in mature lymphocytes may be important factors influencing the homing process of hematopoietic stem cells to the bone marrow. Fucose is an important terminal residue occurring on oligosaccharide chains of cell surfaceexpressed glycoconjugates. 1To whom correspondence should be addressed Fucosylated glycans are involved in cellcell and cellmatrix interactions, differentiation, proliferation, and apoptosis (Varki, 1993). In hematopoietic cells, these glycans play an important role in the recirculation of lymphocytes from the bloodstream into lymphatic organs (Stoolman and Rosen, 1983; Gallatin et al., 1983) and in permitting adhesive contacts between hematopoietic progenitor cells and bone marrow stromal cells (Schmitz et al., 1996; Le Marer and Skacel, 1999). The expression of some fucosylated antigens such as LeX (CD15, Gal14[Fuc-1-3]GlcNAc-) and sialosyl-LeX (CD15s, NeuAc23Gal-1-4[Fuc1-3]GlcNAc-) on hematopoietic cells has been studied (Terstappen et al., 1990; Kannagi, 1997). However, a systematic and comprehensive analysis of the fucosylated histo-blood group antigens LewisY (LeY, CD174 [Histo-blood group antigens H2 and LeY were assigned as CD173 and CD174, respectively, during the Seventh Workshop and Conference on Human Leucocyte Differentiation Antigens in Harrogate in June 2000; Schwartz-Albiez, 2001. In this study we use the new designations for these oligosaccharide structures. For new CD designations please refer to http:// gryphon.jr2.ox.ac.uk/.], Fuc1-2Gal1-4(Fuc1-3)GlcNAc1-), H2 (CD173, Fuc1-2Gal1-4GlcNAc-), H3 (Fuc1-2Gal1-3GalNAc-), and H4 (Fuc1-2Gal1-3GlcNAc1-3Gal1-4Gal1-4Glc-) with regard to their expression on human hematopoietic cells has not yet been done. We have now examined the expression of LeY (CD174), H2 (CD173), H3, and H4 on a broad variety of human leukemia cell lines, isolated lymphocytes from tonsils and peripheral blood, and CD34+ hematopoietic precursors derived from peripheral blood. CD34+ leukemia cell lines such as KG1, KG1a (immature myeloid cell lines), TF1 (immature erythroblastoid cell line), and native CD34+ hematopoietic precursor cells were found to express the CD173 and CD174 antigens. Furthermore, possible carrier molecules of CD173 and CD174 in CD34+ hematopoietic progenitor cells were analyzed. Because the expression of the CD173 antigen is regulated by the -D-galactoside 2--Lfucosyltransferase gene (FUT1), we also tested the presence of FUT1 mRNA by reverse transcription polymerase chain reaction (RT-PCR) in lymphocytes of early and late stages of hematopoietic development. We observed that CD34+ hematopoietic cell lines such as KG1, KG1a, and TF1 expressed the CD173 and CD174 antigens as demonstrated by flow cytometric analysis and immunohistological staining (Figures 1 and 2, Table I). Consequently, native CD34+ hematopoietic precursor cells enriched from peripheral Fig. 1. (A, B) Multicolor flow cytometric analysis of H2 and LeY on CD34+ hematopoietic precursor cells from human peripheral blood. Cells were stained with mAbs specific for H2 (A46-B/B10, IgM) and LeY (A70-C/C8, IgM) followed by incubation with anti-mouse IgM ( -chain) conjugated to FITC (fluorescence 1), anti-CD34 (IgG) conjugated to PE (fluorescence 2) or Cy5 (fluorescence 3), anti-CD33 (IgG), or anti-CD38 (IgG) conjugated to PE (fluorescence 2). (...truncated)


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Yi Cao, Anette Merling, Uwe Karsten, Reinhard Schwartz-Albiez. The fucosylated histo-blood group antigens H type 2 (blood group O, CD173) and Lewis Y (CD174) are expressed on CD34+ hematopoietic progenitors but absent on mature lymphocytes, Glycobiology, 2001, pp. 677-683, 11/8, DOI: 10.1093/glycob/11.8.677