Inhibition of apoptosis by expression of antiapoptotic proteins in recombinant human keratinocytes.
The Fas ligand/Fas interaction plays an important role in the regulation of immune responses. Allografted cells undergo Fas-mediated apoptosis induced by CD8+ T cells. Our objective was to prevent human keratinocytes from immunologically induced apoptosis. We focused on three proteins with inhibitory function on Fas-mediated apoptosis. Human keratinocytes were transfected with either Flip, Faim, or Lifeguard (LFG). The treatment proved to be practicable and efficient. The recombinant keratinocytes with expression of our target proteins were cocultured with CD8+ T cells and the apoptotic activity was then evaluated. Activation of caspase-8 was detectable in control but not in the recombinant cells. Quantitative analysis revealed significant induction of T-cell-induced apoptosis in nontransfected keratinocytes (p = 0.04, n = 12) but not in Flip (p = 0.66), Faim (p = 0.42), or LFG (p = 0.44) expressing cells. Our results suggest that heterotopic expression of antiapoptotic proteins can induce the resistance of keratinocytes to a major mechanism of rejection.
Choi
Claudia Y U
CY
Department of Plastic, Hand and Reconstructive Surgery, Medical School Hannover, D-30625 Hannover, Germany. Choi.Claudia@MH-Hannover.de
Reimers
Kerstin
K
Allmeling
Christina
C
Kall
Susanne
S
Choi
Yeong-Hoon
YH
Vogt
Peter M
PM
D017209:Apoptosis / Q000187:drug effects / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000378:metabolism / Q000494:pharmacology; D016827:CD8 Antigens / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002470:Cell Survival / Q000276:immunology; D002478:Cells, Cultured; D018920:Coculture Techniques; D053222:Fas Ligand Protein / Q000276:immunology; D005240:Feasibility Studies; D015870:Gene Expression; D006084:Graft Rejection / Q000517:prevention & control*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D014162:Transfection; D019014:fas Receptor / Q000276:immunology
D051017:Apoptosis Regulatory Proteins; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D016827:CD8 Antigens; C421307:FAIM protein, human; C400427:FAIM2 protein, human; D053222:Fas Ligand Protein; D008565:Membrane Proteins; D011994:Recombinant Proteins; D019014:fas Receptor