Processing and presentation of intact hen egg-white lysozyme by dendritic cells.
Dendritic cells in lymphoid tissues are of key importance as highly specialized antigen-presenting cells for the induction of T lymphocyte responses. Conflicting results have been published regarding antigen processing of intact proteins by dendritic cells. We now report that highly purified dendritic cells isolated from H-2k mouse spleens very efficiently generated immunogenic fragments of intact hen egg-white lysozyme (HEL) protein to present to an I-Ak-restricted T hybridoma cell line, specific for HEL peptide 46-61. Dendritic cells required 100 times less HEL protein than lipopolysaccharide-induced B cell blasts for effective presentation. Uptake of 125I-labeled HEL protein by dendritic cells and inhibition of presentation of HEL protein by chloroquine treatment was observed. This indicates an endocytotic process and the involvement of acidified compartments. Since the supernatant of dendritic cells, that were incubated with intact HEL protein, contained immunogenic fragments, further evidence for processing of HEL protein by dendritic cells was obtained. When HEL protein was covalently coupled to beads, dendritic cells were not able to ingest these beads, but could still process HEL protein for presentation. This suggests cell surface processing of HEL protein, although internalization of HEL protein released from the beads cannot be excluded. Taken together, these data show that H-2k dendritic cells are capable of processing and presenting intact HEL protein.
De Bruijn
M L
ML
Division of Immunology, The Netherlands Cancer Institute, Amsterdam.
Nieland
J D
JD
Harding
C V
CV
Melief
C J
CJ
D000818:Animals; D000938:Antigen-Presenting Cells / Q000502:physiology*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000502:physiology*; D004527:Egg Proteins / Q000276:immunology* / Q000378:metabolism; D008070:Lipopolysaccharides; D051379:Mice; D008806:Mice, Inbred AKR; D008810:Mice, Inbred C57BL; D009113:Muramidase / Q000276:immunology* / Q000378:metabolism; D010529:Peritoneal Cavity / Q000166:cytology
D004527:Egg Proteins; D008070:Lipopolysaccharides; D009113:Muramidase