Interactions of dendritic cells with fibronectin and endothelial cells.
We studied the phenotypic characteristics of spontaneously migrated skin dendritic cells (sDC) and monocyte-derived dendritic cells (moDC), generated under different culture conditions, and their interactions with fibronectin (FN) and endothelial cells. Monocyte-derived dendritic cells were obtained after culturing monocytes with granulocyte-macrophage colony-stimulating factor (GM-CSF) (800 U/ml) and interleukin-4 (IL-4) (500 U/ml) with either 10% fetal bovine serum (FBS) or 10% allogeneic human serum (HS). Regardless of the type of serum used, the majority of moDC expressed human leucocyte antigen-DR (HLA-DR) and CD86. On day 5 of incubation, 20-67% of moDC cultured in the presence of HS (HS-moDC) expressed CD1a, b and c versus 94-97% when cultured in the presence of FBS (FBS-moDC). DC showed a differential gradient of adhesion to FN: FBS-moDC>HS-moDC>sDC approximately monocytes. Both FBS-moDC and HS-moDC were strongly positive for CD49e (alpha5-integrin) and CD29 (beta1-integrin) but negative for CD49d (alpha4-integrin). A monoclonal antibody (mAb) against CD49e blocked the adhesion of both types of moDC to FN. Although both FBS-moDC and HS-moDC attached to endothelium (a 76% and 63% increase, respectively), only HS-moDC were able to migrate through non-activated endothelium. Overall, these results suggest that spontaneously migrated sDC are less adherent to FN than moDC, that HS and FBS induce differences in CD1 expression, that HS-moDC are less adhesive to FN and endothelial cells but more motile than FBS-moDC, and that alpha5beta1-integrin is the molecule involved in moDC adhesion to FN.
Jancic
C
C
Laboratory of Immunogenetics, Hospital de Clínicas, School of Medicine, University of Buenos Aires, Argentina.
Chuluyan
H E
HE
Morelli
A
A
Larregina
A
A
Kolkowski
E
E
Saracco
M
M
Barboza
M
M
Leiva
W S
WS
Fainboim
L
L
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media; D003713:Dendritic Cells / Q000378:metabolism / Q000502:physiology*; D004727:Endothelium / Q000502:physiology; D005353:Fibronectins / Q000502:physiology*; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016023:Integrins / Q000378:metabolism; D015847:Interleukin-4 / Q000494:pharmacology; D009000:Monocytes / Q000276:immunology*; D012867:Skin / Q000276:immunology*
D003470:Culture Media; D005353:Fibronectins; D016023:Integrins; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor