Engineering mouse T lymphocytes specific to type II collagen by transduction with a chimeric receptor consisting of a single chain Fv and TCR zeta.
The chimeric cell surface receptor scC2Fv/CD8/zeta was constructed to engineer primary mouse T lymphocytes with antibody-type specificity to type II collagen (CII). Such cells could be used as gene carriers in the anti-inflammatory gene therapy of an autoimmune arthritis. This receptor includes the single chain Fv domain (scFv) of the anti-CII monoclonal antibody (mAb) C2, hinge region of CD8alpha and the transmembrane and cytoplasmic domains of TCRzeta. The scC2Fv/CD8/zeta gene was transduced into T cell hybridomas and primary mouse lymphocytes using retrovirus-mediated gene transfer. The chimeric receptor scC2Fv/CD8/zeta forms covalently bound homodimers, as demonstrated in T cell hybridomas and packaging fibroblasts. It does not associate with endogenous signalling subunits of the TCR complex. When scC2Fv/CD8/zeta-expressing clones of T cell hybridomas MD.45 and HCQ6 were stimulated with CII they produced IL-2. The level of their IL-2 response correlated with the expression level of the chimeric receptor on the cell surface. Splenocytes isolated from DBA/1 mice were stimulated with Con A in vitro to facilitate retrovirus-mediated transfer of the scC2Fv/CD8/zeta gene. As a result of transduction, approximately 4% of the Con A-activated splenocytes expressed the chimeric receptor scC2Fv/CD8/zeta on the cell surface. These cells proliferated in response to stimulation with CII.
Annenkov
A
A
Kennedy Institute of Rheumatology, London, UK.
Chernajovsky
Y
Y
D000818:Animals; D001168:Arthritis / Q000628:therapy*; D001327:Autoimmune Diseases / Q000628:therapy*; D003094:Collagen / Q000276:immunology*; D003208:Concanavalin A / Q000494:pharmacology; D015316:Genetic Therapy / Q000379:methods*; D006825:Hybridomas / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics; D007376:Interleukin-2 / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008811:Mice, Inbred DBA; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D013268:Stimulation, Chemical; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection / Q000379:methods*
D007128:Immunoglobulin Fragments; D007376:Interleukin-2; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; C054979:antigen T cell receptor, zeta chain; D003208:Concanavalin A; D003094:Collagen