Formation and kinetics of MHC class I-ovalbumin peptide complexes on immature and mature murine dendritic cells.
Dendritic cells (DC) are professional antigen-presenting cells that are able to induce primary T cell responses. Therefore, several strategies employ peptide-pulsed DC in tumor immunotherapy. For efficient antigen presentation and induction of an immune response by DC the number and stability of MHC I-peptide complexes is crucial. We studied this issue by using the antibody 25-D1.16 that specifically detects OVA peptide SIINFEKL in conjunction with H-2 Kb molecules, and determined its kinetics on mature and immature bone marrow-derived murine DC. Optimal peptide loading was reached after 8-16 h at 50 microM peptide pulse, and was comparable in serum-free versus serum-containing medium. Stimulation of DC with LPS or Poly I:C, and to a lesser extent TNF-alpha, upregulated the total number of surface MHC I molecules and thus improved peptide loading. Pulse-chase experiments revealed a constant half-life of peptide/Kb complexes independent of preceding DC stimulation or their maturation stage. The duration of peptide/Kb complex expression on mature DC, however, could be extended from 24 h to 72 h when the cultures were pretreated with LPS or Poly I:C, but not TNF-alpha. These data might have important implications for the clinical application of peptide-pulsed DC in tumor immunotherapy.
Kukutsch
N A
NA
Department of Dermatology, University of Erlangen-Nuremberg, Erlangen, Germany.
Rossner
S
S
Austyn
J M
JM
Schuler
G
G
Lutz
M B
MB
D000375:Aging / Q000502:physiology*; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism / Q000502:physiology; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism*; D004527:Egg Proteins / Q000378:metabolism*; D006207:Half-Life; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000378:metabolism*; D010446:Peptide Fragments; D013997:Time Factors
D004527:Egg Proteins; D015395:Histocompatibility Antigens Class I; C110366:OVA-8; D010446:Peptide Fragments; D010047:Ovalbumin