Granzyme B and perforin lytic proteins are expressed in CD34+ peripheral blood progenitor cells mobilized by chemotherapy and granulocyte colony-stimulating factor.
Granzyme B and perforin are cytoplasmic granule-associated proteins used by cytotoxic T lymphocytes and natural killer (NK) cells to kill their targets. However, granzyme B gene expression has also been detected in a non-cytotoxic hematopoietic murine multipotent stem cell line, FDCP-Mix. The objective of the present study was to investigate whether granzyme B and perforin could be expressed in human hematopoietic CD34+ cells and if present, discover what their physiologic relevance could be. The primitive CD34+ human cell line KG1a was investigated first and was found to express granzyme B and perforin. Highly purified hematopoietic stem/progenitor cells were then selected using the CD34 surface antigen as marker. Steady-state bone marrow (BM) CD34+ cells did not contain these proteins. Peripheral blood (PB) CD34+ cells, which had been induced to circulate, were also analyzed. After chemotherapy (CT) and granulocyte colony-stimulating factor (G-CSF) treatment, CD34+ cells strongly expressed mRNAs and proteins of granzyme B and perforin. In contrast, CD34+ cells mobilized by G-CSF alone were negative. Western blot analysis further showed that granzyme B and perforin proteins were identical in CD34+ cells and activated PBLs. Such proteins might be implicated in the highly efficient migration of CD34+ stem/progenitor cells from BM to PB after CT and G-CSF treatment. The cellular adhesion mechanisms involved in the BM homing of CD34+ cells are disrupted at least temporarily after CT. The Asp-ase proteolytic activity of granzyme B on extracellular matrix proteins could be used by progenitor cells for their rapid detachment from BM stromal cells and perforin might facilitate their migration across the endothelial cell barrier.
Berthou
C
C
INSERM U 93, Centre de Transfusion sanguine, St Louis Hospital, Paris, France.
Marolleau
J P
JP
Lafaurie
C
C
Soulié
A
A
Dal Cortivo
L
L
Bourge
J F
JF
Benbunan
M
M
Sasportes
M
M
D018952:Antigens, CD34; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D003520:Cyclophosphamide / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D053804:Granzymes; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D017403:In Situ Hybridization; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000473:pathology; D016403:Lymphoma, Large B-Cell, Diffuse; D008228:Lymphoma, Non-Hodgkin / Q000097:blood / Q000188:drug therapy; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D009101:Multiple Myeloma / Q000097:blood / Q000188:drug therapy; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured
D018952:Antigens, CD34; D000970:Antineoplastic Agents; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases