Targeting vectors for intracellular immunisation |
| |
| Institution: | 1. Instituto de Investigaciones Agropecuarias (INIA) Quilamapu, Chillán, Chile;2. Facultad de Agronomía, Universidad de Concepción, Chillán, Chile;1. Central Japan Cord Blood Bank, Seto, Japan;2. Department of Hematology and Oncology, Nakagami Hospital, Okinawa, Japan;3. Department of Promotion for Blood and Marrow Transplantation, Aichi Medical University School of Medicine, Nagakute, Japan;4. Division of Endocrinology, Diabetes and Metabolism, Hematology, Rheumatology (Second Department of Internal Medicine), Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan;5. Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya, Japan;6. Department of Hematology, Shinko Hospital, Kobe, Japan;7. Department of Hematology, Federation of National Public Service Personnel Mutual Aid Associations Toranomon Hospital, Tokyo, Japan;8. Department of Internal Medicine, Kitakyushu Municipal Medical Center, Kitakyushu, Japan;9. Division of Molecular Therapy, The Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan;10. Department of Hematology, Kawasaki Medical School Hospital, Kurashiki, Japan;11. Department of Hematology/Oncology, Tokai University School of Medicine, Isehara, Japan;12. Department of Hematology, Hamanomachi Hospital, Fukuoka, Japan;13. Division of Hematology and Oncology, Department of Medicine, Kurume University Hospital, Kurume, Japan;14. Department of Hematology and Rheumatology, Tohoku University Hospital, Sendai, Japan;15. Department of Hematology/Oncology, Osaka Women''s and Children''s Hospital, Izumi, Japan;16. Japanese Data Center for Hematopoietic Cell Transplantation, Nagoya, Japan;17. Department of Healthcare Administration, Nagoya University Graduate School of Medicine, Nagoya, Japan;18. Hematopoietic Stem Cell Transplantation Division, National Cancer Center Hospital, Tokyo, Japan;19. Department of Hematology and Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan;20. Division of Cell Transplantation and Regenerative Medicine, Department of Innovative Medical Science, Tokai University School of Medicine, Isehara, Japan;21. Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto, Japan |
| |
| Abstract: | We define intracellular immunization as the inhibition or inactivation of the function of a molecule by the ectopic intracellular expression of antibody binding domains which recognise the molecule. Such recombinant antibodies can be directed to different compartments of eukaryotic cells by means of previously defined targeting signals, thus permiting the study of any molecule in any cellular compartment for which an antibody is available. For this purpose, we have created a set of vectors based on the VHExpress vector described Persic, L., Roberts, A., Wilton, J., Cattaneo, A., Bradbury, A. and Hoogenboom, H.R. (1997) An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries. Gene 187, 000–000], which has been modified to express scFvs (single chain fragments) linked to specific targeting signals. These permit the localisation of scFvs to different intracellular compartments: the endoplasmic reticulum (scFvE-er), the nucleus (scFvE-nuclear), the mitochondria (scFvE-mit), the cytoplasm (scFvE-cyto), and as secreted proteins (scFvE-sec). The function of these vectors has been assessed by the immunofluorescence of COS cells transiently transfected with constructs containing the αD11 scFv. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
