Host immune responses in ex vivo approaches to cutaneous gene therapy targeted to keratinocytes.
Epidermal gene therapy may benefit a variety of inherited skin disorders and certain systemic diseases. Both in vivo and ex vivo approaches of gene transfer have been used to target human epidermal stem cells and achieve long-term transgene expression in immunodeficient mouse/human chimera models. Immunological responses however, especially in situations where a neoantigen is expressed, are likely to curtail expression and thereby limit the therapy. In vivo gene transfer to skin has been shown to induce transgene-specific immune responses. Ex vivo gene transfer approaches, where keratinocytes are transduced in culture and transplanted back to patient, however, may avoid signals provided to the immune system by in vivo administration of vectors. In the current study, we have developed a stable epidermal graft platform in immunocompetent mice to analyze host responses in ex vivo epidermal gene therapy. Using green fluorescent protein (GFP) as a neoantigen and an ex vivo retrovirus-mediated gene transfer to mouse primary epidermal cultures depleted of antigen-presenting cells (APCs), we show induction of GFP-specific immune responses leading to the clearance of transduced cells. Similar approach in immunocompetent mice tolerant to GFP resulted in permanent engraftment of transduced cells and continued GFP expression. Activation of transgene-specific immune responses in ex vivo gene transfer targeted to keratinocytes require cross-presentation of transgene product to APCs, a process that is most amenable to immune modulation. This model may be used to explore strategies to divert transgene-specific immune responses to less destructive or tolerogenic ones.
Lu
Z
Z
Department of Dermatology, College of Physicians and Surgeons, Columbia University, NY 10032, USA.
Ghazizadeh
S
S
D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D017931:DNA Primers; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D015603:Keratinocytes / Q000276:immunology / Q000502:physiology* / Q000637:transplantation*; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D023421:Models, Animal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000276:immunology*; D012871:Skin Diseases / Q000235:genetics / Q000276:immunology; D016038:Skin Transplantation / Q000276:immunology*; D014161:Transduction, Genetic
D017931:DNA Primers; D049452:Green Fluorescent Proteins