title: Targeted delivery of tumor necrosis factor-related apoptosis-inducing ligand to keratinocytes with a pemphigus mAb.
abstract: We determined the feasibility of using an anti-desmoglein (Dsg) mAb, Px44, to deliver a biologically active protein to keratinocytes. Recombinantly produced Px44-green fluorescent protein (GFP) injected into mice and skin organ culture delivered GFP to the cell surface of keratinocytes. We replaced GFP with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to produce Px44-TRAIL. We chose TRAIL as a biological model because it inhibits activated lymphocytes and causes apoptosis of hyperproliferative keratinocytes, features of various skin diseases. Px44-TRAIL formed a trimer, the biologically active form of TRAIL. Standard assays of TRAIL activity showed that Px44-TRAIL caused apoptosis of Jurkat cells and inhibited IFN-γ production by activated CD4+ T cells. Enzyme-linked immunoassay with Px44-TRAIL showed delivery of TRAIL to Dsg. Immunofluorescence with Px44-TRAIL incubated on skin sections and cultured keratinocytes or injected into mouse skin, human organ culture, or human xenografts detected TRAIL on keratinocytes. Px44-TRAIL caused apoptosis of the hyperproliferative, but not differentiating, cultured keratinocytes through binding to Dsg3. Foldon, a small trimerization domain, cloned into Px44-TRAIL maintained its stability and biological activity at 37° C for at least 48 hours. These data suggest that such targeted therapy is feasible and may be useful for hyperproliferative and inflamed skin diseases.
authors: Department of Dermatology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
authors: Michiyoshi
authors: M
authors: Kouno
authors: Chenyan
authors: C
authors: Lin
authors: Norman M
authors: NM
authors: Schechter
authors: Don
authors: D
authors: Siegel
authors: Xiaoping
authors: X
authors: Yang
authors: John T
authors: JT
authors: Seykora
authors: John R
authors: JR
authors: Stanley
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D017209:Apoptosis / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D049109:Cell Proliferation; D051184:Desmoglein 3 / Q000276:immunology*; D016503:Drug Delivery Systems / Q000379:methods*; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D015603:Keratinocytes / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques; D010392:Pemphigus / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics*
chemical_list: D000911:Antibodies, Monoclonal; C071894:DSG3 protein, human; D051184:Desmoglein 3; D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; C505654:Tnfsf10 protein, mouse; D049452:Green Fluorescent Proteins