Use of anti-BDCA-2 antibody for detection of dendritic cells type-2 (DC2) in allogeneic hematopoietic stem cell transplantation.
TH2-inducing dendritic cells (DC2) are commonly identified as negative for lineage markers and positive for HLA-DR and CD123 expression. More recently, normal blood DC2 were shown also to be positive for BDCA-2 and BDCA-4 antigens. The aim of this study was to evaluate whether BDCA-2 expression on DC2 is impaired in patients undergoing an allogeneic hematopoietic stem cell transplantation (HSCT) and in healthy donors treated with G-CSF for HSC mobilization. Flow cytometry assays for DC2 detection using either a triple staining with anti-HLA-DR PerCP, anti-Lin(+) anti-CD34 FITC and anti-CD123 PE monoclonal antibodies (mAbs), or a double staining with anti-HLA-DR PE and anti-BDCA-2 FITC mAbs were compared in blood samples from patients who underwent an allogeneic HSCT (n = 30) or from healthy donors before (n = 11) and after (n = 8) G-CSF mobilization, as well as in healthy donors' leukapheresis products (n = 12) or bone marrow (n = 4). Staining of BDCA-2(+) cells with other markers such as anti-CD38, anti-CD54 and anti-CD58 were also performed. Median values of CD123(+) DC2 and BDCA-2(+) DC2 were not statistically different in the blood of patients previously treated with chemotherapy, nor in the blood or bone marrow of heathy donors. Also, a 5 day G-CSF treatment did not affect BDCA-2 or adhesion molecule expression on healthy donors' blood DC2 significantly. A correlation between all the results (n = 65) obtained with the two assays was demonstrated in a linear regression curve (r = 0.914) (P = 0.00001). BDCA-2 is a marker highly specific for DC2 that is not downregulated by chemotherapy or G-CSF treatment. Therefore, the anti-BDCA-2 mAb can be efficiently combined with other mAbs and used in studies addressing the role of DC2 in the allogeneic HSCT setting.
Arpinati
M
M
Research Center for Transplant Immunology, Institute of Hematology and Medical Oncology Seràgnoli, University of Bologna, Bologna, Italy.
Chirumbolo
G
G
Urbini
B
B
Martelli
V
V
Stanzani
M
M
Falcioni
S
S
Bonifazi
F
F
Bandini
G
G
Tura
S
S
Baccarani
M
M
Rondelli
D
D
D000911:Antibodies, Monoclonal; D015415:Biomarkers / Q000032:analysis; D001773:Blood Cells / Q000166:cytology / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology; D016022:Case-Control Studies; D002452:Cell Count / Q000379:methods; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000473:pathology; D005434:Flow Cytometry / Q000379:methods; D019337:Hematologic Neoplasms / Q000097:blood / Q000628:therapy; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006801:Humans; D037181:Lectins, C-Type / Q000276:immunology / Q000378:metabolism*; D007937:Leukapheresis; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D014184:Transplantation, Homologous / Q000379:methods
D000911:Antibodies, Monoclonal; D015415:Biomarkers; C443299:CLEC4C protein, human; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic