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Expression of sialyl-lewisx on CD10-positive normal bone marrow cells
Leukemia Research Vol. 17, No. 4, pp. 389-390, 1993. Printed in Great Britain,
0145-2126/93 $6.00 + .00 ~) 1993 Pergamon Press Ltd
LETTER TO THE EDITORS EXPRESSION
OF SIALYL-LEWIS
x ON CD10-POSITIVE CELLS
NORMAL
BONE
MARROW
(Received 29 October 1992. Revision accepted 7 November 1992)
THE sialyl-Le x antigen is present in a wide range of myeloid cells, natural killer cells, and CD10-positive leukemic lymphoblasts, although it is not expressed on T-cells, B-cells, and CD34-positive normal bone marrow cells [1,2]. It is not clear whether the antigen is detected on CD10-positive normal bone marrow cells, since carbohydrate profiles of the normal lymphoid precursors may differ from those of the leukemic counterparts. We examined the reactivity of an anti-sialyl-Le x monoclonal antibody with CD 10positive normal bone marrow cells. Bone marrow mononuclear cells were collected from three healthy volunteers. The cells were incubated with anti-sialyl-Le x monoclonal antibody (SNH3) or anti-Le y monoclonal antibody (AH6), subsequently with fluorescein isothiocyanate-conjugated anti-mouse IgM. After blocking the free combining sites with mlg, the samples were stained with phycoerythrin (PE)-conjugated anti-CD10 (J5). In another experiment, the cells stained with PEconjugated J5 were incubated with biotinylated antiCD34 (4A1) [3]. The samples were counterstained with allophycocyanin-labeled avidin. Stained cells were measured on a FACStar p~us flow cytometer as described previously [2]. Results were analyzed after setting a gate around the lymphoid/blast cell population. As shown in Fig. 1, a fraction of CD10positive normal bone marrow cells expressed sialylLe x and CD34 but no Le y. In the regenerating bone marrow after chemotherapy, cells co-expressing CD10 and sialyl-Le x were also observed (data not shown). CD34+CD10+CD19 + cells are thought to represent B-cell precursors in normal bone marrow [4]. Since
the sialyl-Le x antigen is not expressed on CD34positive normal bone marrow cells, T-cells and Bcells, it may transiently appear in CD10-positive normal pre-B-cells and may disappear depending on differentiation of the precursors.
REFERENCES 1. Ohmori K., Yoneda T., Ishihara G., Shigeta K., Hirashima K., Kanai M., Itai S., Sasaoki T., Arii S., Arita H. & Kannagi R. (1989) Sialyl SSEA-1 antigen as a carbohydrate marker of human natural killer cells and immature lymphoid cells. Blood 74, 255-261. 2. Muroi K., Suda T., Nojiri H., Ema H., Amemiya Y., Miura Y., Nakauchi H., Singhal A. & Hakomori S. (1992) Reactivity profiles of leukemic myeloblasts with monoclonal antibodies directed to sialosyl-Lex and other lacto-series type 2 chain antigens: absence of reactivity with normal hematopoietic progenitor cells. Blood 79, 713-719. 3. Gunji Y., Nakamura M., Hagiwara T., Hayakawa K., Matsushita H., Osawa H., Nagayoshi K., Nakauchi H., Yanagisawa M., Miura Y. & Suda T. (1992) Expression and function of adhesion molecules on human hematopoietic stem cells: CD34 ÷ LFA-1- cells are more primitive than CD34 ÷ LFA-1 ÷ cells. Blood 80, 429-436. 4. Terstappen L. W. M. M., Huang S., Safford M., Lansdorp P. M. & Loken M. R. (1991) Sequential generations of hematopoietic colonies derived from single-non-lineage-committed CD34 ÷ CD38- progenitor cells. Blood 77, 1218--1227. KAZUO MUROI* MITSURU NAKAMURA* TOSHIO SUDA* YASUSADA MIURA* and SEN-ITIROHHAKOMORIt * Division of Hematology, Dept of Medicine, Jichi Medical School, Tochigi-Ken 329-04, Japan t The Biomembrane Institute and Dept of Pathobiology, University of Washington, Seattle, Washington, U.S.A.
Correspondence to: Kazuo Muroi, M.D., Division of Hematology, Department of Medicine, Jichi Medical School, Minamikawachi-Machi, Tochigi-Ken 329-04, Japan.
389
390
Letter to the Editors
0
0, p -
SLX
LY FIG. 1. FACS-analysis of normal bone marrow mononuclear cells. A fraction of CD10 ÷ cells contains sialylLe x+ cells (A) and CD34 + cells (C) but no Le y+ cells (B). SLX, sialyl-LeX; LY, LeY.
CD34
0145-2126/93 $6.00 + .00 ~) 1993 Pergamon Press Ltd
LETTER TO THE EDITORS EXPRESSION
OF SIALYL-LEWIS
x ON CD10-POSITIVE CELLS
NORMAL
BONE
MARROW
(Received 29 October 1992. Revision accepted 7 November 1992)
THE sialyl-Le x antigen is present in a wide range of myeloid cells, natural killer cells, and CD10-positive leukemic lymphoblasts, although it is not expressed on T-cells, B-cells, and CD34-positive normal bone marrow cells [1,2]. It is not clear whether the antigen is detected on CD10-positive normal bone marrow cells, since carbohydrate profiles of the normal lymphoid precursors may differ from those of the leukemic counterparts. We examined the reactivity of an anti-sialyl-Le x monoclonal antibody with CD 10positive normal bone marrow cells. Bone marrow mononuclear cells were collected from three healthy volunteers. The cells were incubated with anti-sialyl-Le x monoclonal antibody (SNH3) or anti-Le y monoclonal antibody (AH6), subsequently with fluorescein isothiocyanate-conjugated anti-mouse IgM. After blocking the free combining sites with mlg, the samples were stained with phycoerythrin (PE)-conjugated anti-CD10 (J5). In another experiment, the cells stained with PEconjugated J5 were incubated with biotinylated antiCD34 (4A1) [3]. The samples were counterstained with allophycocyanin-labeled avidin. Stained cells were measured on a FACStar p~us flow cytometer as described previously [2]. Results were analyzed after setting a gate around the lymphoid/blast cell population. As shown in Fig. 1, a fraction of CD10positive normal bone marrow cells expressed sialylLe x and CD34 but no Le y. In the regenerating bone marrow after chemotherapy, cells co-expressing CD10 and sialyl-Le x were also observed (data not shown). CD34+CD10+CD19 + cells are thought to represent B-cell precursors in normal bone marrow [4]. Since
the sialyl-Le x antigen is not expressed on CD34positive normal bone marrow cells, T-cells and Bcells, it may transiently appear in CD10-positive normal pre-B-cells and may disappear depending on differentiation of the precursors.
REFERENCES 1. Ohmori K., Yoneda T., Ishihara G., Shigeta K., Hirashima K., Kanai M., Itai S., Sasaoki T., Arii S., Arita H. & Kannagi R. (1989) Sialyl SSEA-1 antigen as a carbohydrate marker of human natural killer cells and immature lymphoid cells. Blood 74, 255-261. 2. Muroi K., Suda T., Nojiri H., Ema H., Amemiya Y., Miura Y., Nakauchi H., Singhal A. & Hakomori S. (1992) Reactivity profiles of leukemic myeloblasts with monoclonal antibodies directed to sialosyl-Lex and other lacto-series type 2 chain antigens: absence of reactivity with normal hematopoietic progenitor cells. Blood 79, 713-719. 3. Gunji Y., Nakamura M., Hagiwara T., Hayakawa K., Matsushita H., Osawa H., Nagayoshi K., Nakauchi H., Yanagisawa M., Miura Y. & Suda T. (1992) Expression and function of adhesion molecules on human hematopoietic stem cells: CD34 ÷ LFA-1- cells are more primitive than CD34 ÷ LFA-1 ÷ cells. Blood 80, 429-436. 4. Terstappen L. W. M. M., Huang S., Safford M., Lansdorp P. M. & Loken M. R. (1991) Sequential generations of hematopoietic colonies derived from single-non-lineage-committed CD34 ÷ CD38- progenitor cells. Blood 77, 1218--1227. KAZUO MUROI* MITSURU NAKAMURA* TOSHIO SUDA* YASUSADA MIURA* and SEN-ITIROHHAKOMORIt * Division of Hematology, Dept of Medicine, Jichi Medical School, Tochigi-Ken 329-04, Japan t The Biomembrane Institute and Dept of Pathobiology, University of Washington, Seattle, Washington, U.S.A.
Correspondence to: Kazuo Muroi, M.D., Division of Hematology, Department of Medicine, Jichi Medical School, Minamikawachi-Machi, Tochigi-Ken 329-04, Japan.
389
390
Letter to the Editors
0
0, p -
SLX
LY FIG. 1. FACS-analysis of normal bone marrow mononuclear cells. A fraction of CD10 ÷ cells contains sialylLe x+ cells (A) and CD34 + cells (C) but no Le y+ cells (B). SLX, sialyl-LeX; LY, LeY.
CD34