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Published on: March 17, 2014
CDCP1 is a novel marker for hematopoietic stem cells
Tim Conze1, Reiner Lammers, Selim Kuci
1Department of Internal Medicine II, Division of Hematology, Immunology, and Oncology, University of Tübingen, Tübingen, Germany.
Insights
Researchers identified CDCP1 as a novel marker for hematopoietic stem cells. This transmembrane protein is highly expressed on a subset of CD34(+) stem/progenitor cells in bone marrow.
Area of Science:
- Cell Biology
- Hematology
- Immunology
Background:
- CDCP1 (CUB-domain containing protein 1) is a transmembrane protein.
- CDCP1 mRNA shows high expression in lung and colon tumors and K562 cells.
- The extracellular domain features three CUB domains, and the cytoplasmic region has a hexalysine stretch.
Purpose of the Study:
- To develop monoclonal antibodies for analyzing CDCP1 protein expression.
- To identify the cell types expressing CDCP1 in peripheral blood and bone marrow.
- To investigate the potential of CDCP1 as a marker for hematopoietic stem cells.
Main Methods:
- Overexpression of CDCP1 in NIH-3T3 cells for antibody generation.
- Immunization of Balb/c mice and generation of hybridoma clones.
- Multi-color flow cytometry analysis of peripheral blood and bone marrow cells.
- Transplantation of purified CDCP1(+) cells into NOD/SCID mice.
Main Results:
- Four monoclonal antibodies (CUB1-CUB4) were successfully generated against CDCP1.
- CDCP1 protein was not detected on mature blood cells (lymphocytes, monocytes, granulocytes, erythrocytes, platelets).
- CDCP1 is predominantly expressed on a subset of CD34(+) stem/progenitor cells in bone marrow.
- Transplanted CDCP1(+) cells engrafted and showed multi-lineage differentiation potential.
Conclusions:
- CDCP1 is a novel marker for identifying hematopoietic stem cells.
- CDCP1 expression is restricted to a specific population of stem/progenitor cells.
- These findings have implications for stem cell research and potential therapeutic applications.
Abstract:
CDCP1 is a transmembrane protein that contains three CUB domains within the extracellular region and a hexalysine stretch within the cytoplasmic region. CDCP1 mRNA is highly expressed in lung and colon tumors and in the erythroleukemic cell line K562. To analyze CDCP1 protein expression, monoclonal antibodies against the extracellular domain of CDCP1 were raised. For this purpose, CDCP1 was overexpressed in NIH-3T3 cells. Balb/c mice were then immunized with the resultant cell line NIH-3T3/huCDCP1. After fusion of SP2/0 cells with immune spleen cells, hybridoma clones were selected that secreted antibodies reacting with NIH-3T3/huCDCP1 cells but not with parental cells. Four antibodies (CUB1-CUB4) were obtained that fulfilled these criteria. Screening of peripheral blood cells revealed that the antibodies did not recognize mature lymphocytes, monocytes, granulocytes, erythrocytes, or platelets. In contrast, multi-color analyses revealed that CDCP1 protein is almost exclusively expressed on a subset of CD34(+) stem/progenitor cells in bone marrow. Transplantation of purified CDCP1(+) cells into NOD/SCID mice resulted in engraftment of human cells with multi-lineage differentiation potential, suggesting that CDCP1 is a novel marker for hematopoietic stem cells.
