Cross-presentation of a CMV pp65 epitope by human dendritic cells using bee venom PLA2 as a membrane-binding vector.
We have used bee venom phospholipase A2 as a vector to load human dendritic cells ex vivo with a major histocompatibility complex (MHC) class I-restricted epitope fused to its C-terminus. The fusion protein bound to human monocyte-derived dendritic cells and was internalized into early endosomes. In vitro immunization experiments showed that these dendritic cells were able to generate specific CD8 T cell lines against the epitope carried by the fusion protein. Cross-presentation did not require proteasome, transporter associated with antigen processing, or endosome proteases, but required newly synthesized MHC molecules. Comparison of the antigen presentation pathway observed in this study to that followed by other toxins used as vectors is discussed.
Babon
Aurélie
A
Protein Engineering and Research Department (DIEP), bat 152, CEA-Saclay, 91191 Gif sur Yvette cedex, France.
Almunia
Christine
C
Boccaccio
Claire
C
Beaumelle
Bruno
B
Gelb
Michael H
MH
Ménez
André
A
Maillère
Bernard
B
Abastado
Jean-Pierre
JP
Salcedo
Margarita
M
Gillet
Daniel
D
D017951:Antigen Presentation / Q000187:drug effects / Q000502:physiology; D000956:Antigens, Viral / Q000276:immunology*; D001514:Bee Venoms / Q000201:enzymology; D020126:Brefeldin A / Q000494:pharmacology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002462:Cell Membrane / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D045142:Cross-Priming / Q000187:drug effects; D003713:Dendritic Cells / Q000737:chemistry / Q000276:immunology* / Q000502:physiology; D011992:Endosomes / Q000502:physiology; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D010741:Phospholipases A / Q000032:analysis / Q000378:metabolism*; D054467:Phospholipases A2; D010750:Phosphoproteins / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000502:physiology; D061988:Proteasome Inhibitors; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000032:analysis / Q000276:immunology* / Q000378:metabolism
D000956:Antigens, Viral; D001514:Bee Venoms; D015395:Histocompatibility Antigens Class I; D016056:Immunodominant Epitopes; D010750:Phosphoproteins; D061988:Proteasome Inhibitors; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D020126:Brefeldin A; D002738:Chloroquine; D010741:Phospholipases A; D054467:Phospholipases A2; D046988:Proteasome Endopeptidase Complex