MHC class I manipulation on cell surfaces by gene transfer of anti-MHC class I intrabodies—a tool for decreased immunogenicity of allogeneic tissue and cell transplants

• Published in: Methods (San Diego, Calif.) Vol. 34; no. 2; pp. 240 - 249
• Main Authors: Busch, Annette, Marasco, Wayne A, Doebis, Cornelia, Volk, Hans-Dieter, Seifert, Martina
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2004
• Subjects: Animals; Antibodies - genetics; Antibodies - metabolism; Antibodies intracellular; Antigens, Surface - genetics; Antigens, Surface - immunology; Bicistronic expression; Cell Line; Cell Membrane - genetics; Cell Membrane - immunology; Cell Membrane - metabolism; Cell Transplantation - methods; Cells, Cultured; Fusionprotein; Gene Transfer Techniques; Genes, MHC Class I; Humans; Intrabodies; Keratinocytes - immunology; Major histocompatibility antigen class I; Rats; Tissue engineering; Transfection; Transplantation, Homologous - immunology; Major histocompatibility antigen class I; Transfection; Tissue engineering; Antibodies intracellular; Bicistronic expression; Fusionprotein; Intrabodies
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2004.03.017

Intrabodies (IB) are suitable tools to down-regulate the expression of cell surface molecules in general. In this work, the appearance of major histocompatibility (MHC) class I molecules on the cell surface could be prevented by the expression of intracellularly localized anti-MHC class I antibodies. The expression of MHC antigens presenting intracellularly synthetised peptides on the cell surface is the predominant reason for immunologic detection and rejection of allogeneic cell and tissue transplants. Allogeneic keratinocyte sheets might be a suitable tool for skin grafting. Within this study primary rat keratinocytes have been transfected with anti-MHC I-IB. Strong IB-expressing cells showed a MHC I “knockout” phenotype. The cells did not exhibit any significant alterations compared to non-transfected cells: the cell growth and the expression of other surface molecules were unaltered. Merely an enhanced intracellular accumulation of MHC I molecules could be detected. Notably, IB-expressing keratinocytes displayed a reduced susceptibility to allogeneic cytotoxic T cells in vitro compared to unmodified cells with a normal level of MHC I surface expression. These MHC I-deficient keratinocytes might be utilized in tissue-engineered allogeneic non-immunogeneic skin transplants. The principle of MHC class I manipulation in general can be used for other allogeneic cell and tissue-engineered transplants as well.