title: Characterization of canine monocyte-derived dendritic cells with phenotypic and functional differentiation.
abstract: For therapeutic purposes, large numbers of dendritic cells (DCs) are essential. In this study, we used 2% autologous canine plasma, granulocyte/macrophage colony-stimulating factor (GM-CSF), fms-like tyrosine kinase 3 ligand (Flt3L), and interleukin 4 (IL-4) in generating monocyte-derived DCs from peripheral blood mononuclear cells of dogs. The plasma enriched the population of CD14-positive monocytes by greatly enhancing the efficiency of monocyte adherence, the proportion of adherent cells increasing from 6.6% with 10% fetal bovine serum to 15.3% with 2% autologous canine plasma. Culturing the adherent monocytes for 6 d with human GM-CSF, canine IL-4, and human Flt3L significantly increased the yield of DCs, more than 90% of which were CD14-negative. Because, in the presence of lipopolysaccharide (LPS), monocytes that were CD14-positive expressed tumor necrosis factor ac much more than DCs with low levels of CD14, it is important to decrease the numbers of CD14-positive cells in generating monocyte-derived DCs. With flow cytometry and real-time reverse-transcriptase-mediated polymerase chain reaction assays, we found that in canine immature DCs (iDCs) the expression of DLA class II molecules, CD1a, CD11c, CD40, and CD86 was high and the expression of CD80, CD83, and CD14 either low or negative. During maturation (stimulated by LPS), the expression of CDla, CD40, CD83, and CD80 was upregulated. However, the expression of DLA class II molecules, CD11c, and CD86 was not increased in mature DCs. Incubating the iDCs with LPS decreased antigen uptake and increased the cells' immunostimulatory capacity (assessed by the allogeneic mixed-lymphocyte reaction), indicating that LPS accelerates the functional maturation of DCs. This protocol may facilitate the use of DCs in cellular immunotherapy.
authors: Department of Veterinary Medicine, National Taiwan University.
authors: Yu-Shan
authors: YS
authors: Wang
authors: Kwan-Hwa
authors: KH
authors: Chi
authors: Kuang-Wen
authors: KW
authors: Liao
authors: Cheng-Chi
authors: CC
authors: Liu
authors: Chiao-Lei
authors: CL
authors: Cheng
authors: Yi-Chun
authors: YC
authors: Lin
authors: Chiung-Hsiang
authors: CH
authors: Cheng
authors: Rea-Min
authors: RM
authors: Chu
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018949:Antigens, CD1 / Q000032:analysis; D018122:B7-1 Antigen / Q000032:analysis; D051940:B7-2 Antigen / Q000032:analysis; D019013:CD40 Antigens / Q000032:analysis; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000502:physiology*; D004285:Dogs; D005434:Flow Cytometry / Q000662:veterinary; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006684:HLA-DR Antigens / Q000032:analysis*; D007136:Immunoglobulins / Q000032:analysis; D015847:Interleukin-4 / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D007959:Lymphocyte Culture Test, Mixed; D008562:Membrane Glycoproteins / Q000032:analysis; D009000:Monocytes / Q000166:cytology* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D051941:fms-Like Tyrosine Kinase 3 / Q000494:pharmacology; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D018949:Antigens, CD1; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D019013:CD40 Antigens; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006684:HLA-DR Antigens; D007136:Immunoglobulins; D015847:Interleukin-4; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D051941:fms-Like Tyrosine Kinase 3; D000097802:CD83 Antigen; C419393:CD1a antigen
references: Proc Natl Acad Sci U S A. 1990 Oct;87(19):7698-702
references: 2145580
references: J Vet Med Sci. 2003 Jun;65(6):663-9
references: 12867724
references: Annu Rev Immunol. 1991;9:271-96
references: 1910679
references: Nature. 1992 Nov 19;360(6401):258-61
references: 1279441
references: J Exp Med. 1992 Dec 1;176(6):1693-702
references: 1460426
references: J Exp Med. 1994 Apr 1;179(4):1109-18
references: 8145033
references: J Exp Med. 1994 Jul 1;180(1):83-93
references: 8006603
references: Tissue Antigens. 1994 Mar;43(3):137-54
references: 8091414
references: Tissue Antigens. 1994 Mar;43(3):179-83
references: 7916497
references: Exp Hematol. 1995 Dec;23(14):1463-71
references: 8542932
references: Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2588-92
references: 8637918
references: J Immunol Methods. 1996 Sep 27;196(2):121-35
references: 8841451
references: J Immunol Methods. 1996 Sep 27;196(2):137-51
references: 8841452
references: Vet Immunol Immunopathol. 1996 Oct;53(3-4):329-34
references: 8969052
references: Blood. 1997 Nov 1;90(9):3245-87
references: 9345009
references: Nature. 1998 Mar 5;392(6671):86-9
references: 9510252
references: J Clin Invest. 1999 Jul;104(2):173-80
references: 10411546
references: J Immunol Methods. 2005 Mar;298(1-2):175-82
references: 15847807
references: Vet Immunol Immunopathol. 2006 Jan 15;109(1-2):167-76
references: 16202456
references: J Exp Med. 1999 Dec 6;190(11):1669-78
references: 10587357
references: Cancer Res. 1999 Dec 15;59(24):6028-32
references: 10626784
references: J Immunol. 2000 Feb 1;164(3):1269-76
references: 10640740
references: J Clin Invest. 2000 Mar;105(6):R9-R14
references: 10727452
references: Microbes Infect. 2000 Mar;2(3):257-72
references: 10758402
references: Annu Rev Immunol. 2000;18:767-811
references: 10837075
references: Transplantation. 2000 Nov 27;70(10):1437-42
references: 11118086
references: Nat Immunol. 2000 Aug;1(2):156-62
references: 11248809
references: Science. 1990 Sep 21;249(4975):1431-3
references: 1698311
references: Methods. 2001 Dec;25(4):402-8
references: 11846609
references: J Comp Pathol. 2002 Feb-Apr;126(2-3):238-41
references: 11945016
references: J Immunol. 2002 May 1;168(9):4333-43
references: 11970975
references: ANZ J Surg. 2002 Jul;72(7):501-6
references: 12123513
references: Mol Ther. 2002 Sep;6(3):407-14
references: 12231178
references: Cell Immunol. 2002 Dec;220(2):125-33
references: 12657247
references: Anticancer Res. 2003 Jan-Feb;23(1A):315-21
references: 12680230
references: Vet Immunol Immunopathol. 2003 May 12;92(3-4):149-62
references: 12730015
references: Vet Immunol Immunopathol. 2003 May 30;93(1-2):9-20
references: 12753771
references: J Immunol. 2003 Jun 15;170(12):5861-8
references: 12794111
references: Curr Opin Hematol. 2003 Jul;10(4):272-8
references: 12799532
references: Eur J Immunol. 1990 Nov;20(11):2375-81
references: 1701391