Bacterial lipopolysaccharide contamination of commercial collagen preparations may mediate dendritic cell maturation in culture.
Dendritic cells (DC) are potent antigen presenting cells, which are responsible for the initiation of naive T and T-dependent immune responses. The present studies were based upon recent reports that commercial collagen I preparations induce the maturation of human DC in vitro. We show that human blood monocyte-derived (GM-CSF and IL-4 cultured) DC pulsed on collagen I-coated plates undergo a dose-dependent increase in stimulatory capacity in oxidative mitogenesis assays. This is accompanied by the upregulation of costimulatory molecules (CD40, CD80, CD86), CD25, ICAM-1 and the DC-specific marker CD83. The maturation effect is more potent than TNF-alpha, which is a known mediator of DC function. However, bacterial lipopolysaccharide (LPS), a powerful inducer of DC maturation, was found to be present at very high levels in one commercial collagen solution that was tested. The effect of LPS upon DC maturation was similar to culture with collagen. Furthermore, a different collagen I preparation with low levels of LPS contamination was less effective at inducing DC maturation, while spiking the collagen solution with LPS prior to plastic coating equalised these effects. Finally, human monocyte-derived DC were found not to express typical collagen receptors VLA-1, 2 and 3. We therefore propose that LPS contamination may at least partially explain reported collagen I induced DC maturation.
Nuffield Department of Surgery, University of Oxford John Radcliffe Hospital, UK.
R M
RM
Suri
J M
JM
Austyn
D002478:Cells, Cultured; D003094:Collagen / Q000302:isolation & purification / Q000494:pharmacology*; D003470:Culture Media; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D019012:Integrin beta1 / Q000096:biosynthesis; D016023:Integrins / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology*; D010641:Phenotype; D038981:Receptors, Collagen
D003470:Culture Media; D019012:Integrin beta1; D016023:Integrins; D008070:Lipopolysaccharides; D038981:Receptors, Collagen; D003094:Collagen