Characterization of canine monocyte-derived dendritic cells with phenotypic and functional differentiation.
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.
Wang
Yu-Shan
YS
Department of Veterinary Medicine, National Taiwan University.
Chi
Kwan-Hwa
KH
Liao
Kuang-Wen
KW
Liu
Cheng-Chi
CC
Cheng
Chiao-Lei
CL
Lin
Yi-Chun
YC
Cheng
Chiung-Hsiang
CH
Chu
Rea-Min
RM
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
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
Proc Natl Acad Sci U S A. 1990 Oct;87(19):7698-702
2145580
J Vet Med Sci. 2003 Jun;65(6):663-9
12867724
Annu Rev Immunol. 1991;9:271-96
1910679
Nature. 1992 Nov 19;360(6401):258-61
1279441
J Exp Med. 1992 Dec 1;176(6):1693-702
1460426
J Exp Med. 1994 Apr 1;179(4):1109-18
8145033
J Exp Med. 1994 Jul 1;180(1):83-93
8006603
Tissue Antigens. 1994 Mar;43(3):137-54
8091414
Tissue Antigens. 1994 Mar;43(3):179-83
7916497
Exp Hematol. 1995 Dec;23(14):1463-71
8542932
Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2588-92
8637918
J Immunol Methods. 1996 Sep 27;196(2):121-35
8841451
J Immunol Methods. 1996 Sep 27;196(2):137-51
8841452
Vet Immunol Immunopathol. 1996 Oct;53(3-4):329-34
8969052
Blood. 1997 Nov 1;90(9):3245-87
9345009
Nature. 1998 Mar 5;392(6671):86-9
9510252
J Clin Invest. 1999 Jul;104(2):173-80
10411546
J Immunol Methods. 2005 Mar;298(1-2):175-82
15847807
Vet Immunol Immunopathol. 2006 Jan 15;109(1-2):167-76
16202456
J Exp Med. 1999 Dec 6;190(11):1669-78
10587357
Cancer Res. 1999 Dec 15;59(24):6028-32
10626784
J Immunol. 2000 Feb 1;164(3):1269-76
10640740
J Clin Invest. 2000 Mar;105(6):R9-R14
10727452
Microbes Infect. 2000 Mar;2(3):257-72
10758402
Annu Rev Immunol. 2000;18:767-811
10837075
Transplantation. 2000 Nov 27;70(10):1437-42
11118086
Nat Immunol. 2000 Aug;1(2):156-62
11248809
Science. 1990 Sep 21;249(4975):1431-3
1698311
Methods. 2001 Dec;25(4):402-8
11846609
J Comp Pathol. 2002 Feb-Apr;126(2-3):238-41
11945016
J Immunol. 2002 May 1;168(9):4333-43
11970975
ANZ J Surg. 2002 Jul;72(7):501-6
12123513
Mol Ther. 2002 Sep;6(3):407-14
12231178
Cell Immunol. 2002 Dec;220(2):125-33
12657247
Anticancer Res. 2003 Jan-Feb;23(1A):315-21
12680230
Vet Immunol Immunopathol. 2003 May 12;92(3-4):149-62
12730015
Vet Immunol Immunopathol. 2003 May 30;93(1-2):9-20
12753771
J Immunol. 2003 Jun 15;170(12):5861-8
12794111
Curr Opin Hematol. 2003 Jul;10(4):272-8
12799532
Eur J Immunol. 1990 Nov;20(11):2375-81
1701391