In vitro processing and presentation of a lipidated cytotoxic T-cell epitope derived from measles virus fusion protein.
Lipopeptidic formulations have been described as efficient activators of cytotoxic T lymphocytes (CTL). To better understand the pathway via which lipopeptides reach the MHC class I molecules we studied the intracellular processing and presentation of a measles virus-derived CTL epitope, to which a palmitoyl moiety was added synthetically. The palmitoyl group was conjugated to the N-terminus either directly or via a spacer sequence. The use of single or double fluorescent-labeled lipopeptides allowed the visualization of intracellular processing of these antigens using confocal microscopy. Our data indicate that the spacer composition influences internalization of the conjugate into the cell, proteasomal degradation, translocation into the ER by the transporter associated with antigen processing (TAP), and the intracellular trafficking of lipopeptides.
Stittelaar
K J
KJ
Laboratory for Vaccine Research, National Institute of Public Health and the Environment, 3720 BA, Bilthoven, The Netherlands.
Hoogerhout
P
P
Ovaa
W
W
van Binnendijk
R R
RR
Poelen
M C
MC
Roholl
P
P
van Els
C A
CA
Osterhaus
A D
AD
Wiertz
E J
EJ
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D017951:Antigen Presentation; D020126:Brefeldin A / Q000494:pharmacology; D002461:Cell Line, Transformed / Q000276:immunology; D002478:Cells, Cultured / Q000276:immunology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D006056:Golgi Apparatus / Q000378:metabolism; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D008074:Lipoproteins / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D008459:Measles virus / Q000276:immunology*; D008823:Micelles; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009097:Multienzyme Complexes / Q000378:metabolism; D019308:Palmitic Acid / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D046988:Proteasome Endopeptidase Complex; D021381:Protein Transport / Q000187:drug effects; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D020298:U937 Cells / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000276:immunology*
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters; D000939:Epitopes; D006680:HLA Antigens; D008074:Lipoproteins; D008823:Micelles; D009097:Multienzyme Complexes; D010446:Peptide Fragments; C073119:TAP1 protein, human; D014409:Tumor Necrosis Factor-alpha; D014760:Viral Fusion Proteins; C071692:TAP2 protein, human; D020126:Brefeldin A; D019308:Palmitic Acid; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex