Targeted delivery of peptide epitopes to class I major histocompatibility molecules by a modified Pseudomonas exotoxin.
Cytotoxic T lymphocytes (CTLs) expressing the CD8 surface marker recognize peptides in association with major histocompatibility complex (MHC) class I molecules. Although most peptides expressed on MHC class I molecules are derived from self- or virally encoded proteins, delivery of exogenous proteins to the cytosol can result in their being processed for presentation to CTLs on MHC class I molecules. We describe two fusion proteins (PEMa and PENP), consisting of the binding and translocating domains of Pseudomonas exotoxin A (PE), fused to peptide epitopes from influenza A matrix protein and nucleoprotein, respectively. These fusion proteins were internalized and processed by MHC class I-positive target cells, resulting in sensitization of target cells for lysis by peptide-specific CTLs. A point mutation known to interfere with intoxication by wild-type PE also reduced the ability of PEMa to sensitize target cells. Fusion of peptide or polypeptide epitopes with PE provides a potential means of eliciting CTLs without the use of self-replicating agents, as well as a useful probe for studying MHC class I-restricted antigen processing.
Donnelly
J J
JJ
Department of Cancer Research, Merck Research Laboratories, West Point, PA 19486.
Ulmer
J B
JB
Hawe
L A
LA
Friedman
A
A
Shi
X P
XP
Leander
K R
KR
Shiver
J W
JW
Oliff
A I
AI
Martinez
D
D
Montgomery
D
D
D036002:ADP Ribose Transferases; D000595:Amino Acid Sequence; D000818:Animals; D001427:Bacterial Toxins; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D003062:Codon / Q000235:genetics; D003602:Cytotoxicity, Immunologic; D004279:DNA, Viral / Q000235:genetics; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005098:Exotoxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D009980:Influenza A virus / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D012516:Osteosarcoma; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D013154:Spleen / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D003062:Codon; D004279:DNA, Viral; D000939:Epitopes; D005098:Exotoxins; D005982:Glutathione Transferase; D015395:Histocompatibility Antigens Class I; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A