Specific immunotherapy by genetically engineered APCs: the "guided missile" strategy.
We tested the hypothesis that APCs genetically engineered to present an Ag and to express Fas ligand (FasL) simultaneously can target and eliminate Ag-specific T cells. Transgenic T cells specific for influenza hemagglutinin (HA) were used as targets. We prepared recombinant vaccinia virus vectors (VVV) to transfer the gene constructs individually or simultaneously into APCs. We prevented unwanted viral replication by attenuating the VVVs with psoralen-UV light treatment. For presentation of the HA Ag, APCs were transduced with cDNA for HA flanked by sequences of the lysosome-associated membrane protein that direct efficient processing and presentation of the Ag by APCs. As a "warhead" for the APCs, we transduced them with the gene for FasL, which induces apoptosis of Fas-expressing activated T cells. To protect the transduced APCs from self-destruction by FasL, we transferred cDNA for a truncated form of Fas-associated death domain, which inhibits Fas-mediated cell death. Our results show that the engineered APCs effectively expressed the genes of interest. APCs transduced with VVV carrying all three gene constructs specifically killed HA-transgenic T cells in culture. Coculture with T cells specific for an unrelated Ag (OVA) had no significant effect. Our in vitro findings show that APCs can be genetically engineered to target and kill Ag-specific T cells and represent a promising novel strategy for the specific treatment of autoimmune diseases.
Neuromuscular Research Laboratory, Department of Neurology, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA.
B
B
Wu
J M
JM
Wu
A
A
Miagkov
R N
RN
Adams
H I
HI
Levitsky
D B
DB
Drachman
D048868:Adaptor Proteins, Signal Transducing; D000276:Adjuvants, Immunologic / Q000235:genetics; D019264:Adoptive Transfer / Q000379:methods*; D000818:Animals; D000938:Antigen-Presenting Cells / Q000637:transplantation*; D017209:Apoptosis / Q000235:genetics / Q000276:immunology; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D053222:Fas Ligand Protein; D053200:Fas-Associated Death Domain Protein; D018390:Gene Targeting / Q000379:methods; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000138:chemical synthesis / Q000276:immunology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D006801:Humans; D007399:Interphase / Q000235:genetics / Q000276:immunology; D008024:Ligands; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000633:toxicity; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology; D019014:fas Receptor / Q000235:genetics / Q000276:immunology
D048868:Adaptor Proteins, Signal Transducing; D000276:Adjuvants, Immunologic; D002352:Carrier Proteins; C505533:FADD protein, human; C505661:FASLG protein, human; C505534:Fadd protein, mouse; D053222:Fas Ligand Protein; D053200:Fas-Associated Death Domain Protein; C505658:Fasl protein, mouse; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008024:Ligands; D008562:Membrane Glycoproteins; D019014:fas Receptor