Electroporation and commercial liposomes efficiently deliver soluble protein into the MHC class I presentation pathway. Priming in vitro and in vivo for class I-restricted recognition of soluble antigen.
Class I major histocompatibility complex (MHC)-restricted cytotoxic T lymphocyte responses to ovalbumin (OVA) were evaluated following delivery of soluble antigen mixed with commercial liposomes or by electroporation of soluble protein into target cells. Splenic antigen presenting cells (APC) and transfected L cell lines were sensitised for recognition by OVA-specific, class I-restricted T hybridomas when antigen was introduced by either method into live cells. Delivery of soluble OVA by both electroporation and commercial liposomes proved more efficient than osmotic loading in sensitising for class I presentation. OVA-specific cytotoxic T lymphocytes (CTL) were effectively primed in naive mice following reinjection of spleen cells pulsed with soluble OVA encapsulated by liposomes or electroporated in vitro. These CTL recognised the well defined OVA257-264 determinant in association with H-2Kb and were derived under conditions where CTL activity obtained from cross priming by soluble OVA alone was undetectable. In addition, using electroporation and commercial liposomes, the loading of APC for OVA recognition required intact MHC-linked antigen presentation genes deleted in the T2-Kb cell line. Antigen delivery to APC by electroporation and commercial liposomes provides a simple and efficient way of studying class I-restricted T cell recognition of soluble protein antigens.
Chen
W
W
Department of Clinical Immunology, Flinders Medical Centre, Bedford Park, South Australia.
Carbone
F R
FR
McCluskey
J
J
D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism; D002463:Cell Membrane Permeability; D004337:Drug Carriers; D000939:Epitopes / Q000378:metabolism; D006183:H-2 Antigens / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D006825:Hybridomas; D008081:Liposomes; D008285:Major Histocompatibility Complex; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D000939:Epitopes; D006183:H-2 Antigens; D015395:Histocompatibility Antigens Class I; D008081:Liposomes; D010047:Ovalbumin