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1、of November 30, 2011 This information is current as2000;164;4635-4640 J ImmunolUlmer Gillis R. Otten, Kent Thudium, Mark J. Selby and Jeffrey B. Goldbeck, Susan W. Barnett, Minchao Chen, Louisa Leung, Georg Widera, Melissa Austin, Dietmar Rabussay, CherylImmunogenicity by Electroporation In Vivo Inc
2、reased DNA Vaccine Delivery and Referenceshttp:/www.jimmunol.org/content/164/9/4635.full.html#related-urls Article cited in: http:/www.jimmunol.org/content/164/9/4635.full.html#ref-list-1 , 13 of which can be accessed free at: cites 29 articles This article Subscriptionshttp:/www.jimmunol.org/subscr
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4、 Print ISSN: 0022-1767 Online ISSN: 1550-6606. Immunologists, Inc. All rights reserved. by The American Association of Copyright 2000 9650 Rockville Pike, Bethesda, MD 20814-3994. The American Association of Immunologists, Inc.,is published twice each month by The Journal of Immunologyon November 30
5、, 2011 www.jimmunol.org Downloaded from Increased DNA Vaccine Delivery and Immunogenicity by Electroporation In Vivo Georg Widera, Melissa Austin, Dietmar Rabussay, Cheryl Goldbeck,* Susan W. Barnett,* Minchao Chen,* Louisa Leung,* Gillis R. Otten,* Kent Thudium,* Mark J. Selby,* and Jeffrey B. Ulme
6、r 1 * DNA vaccines have been demonstrated to be potent in small animals but are less effective in primates. One limiting factor may be inefcient uptake of DNA by cells in situ. In this study, we evaluated whether cellular uptake of DNA was a signicant barrier to efcient transfection in vivo and subs
7、equent induction of immune responses. For this purpose, we used the technique of electroporation to facilitate DNA delivery in vivo. This technology was shown to substantially increase delivery of DNA to cells, resulting in increased expression and elevated immune responses. The potency of a weakly
8、immunogenic hepatitis B surface Ag DNA vaccine was increased in mice, as seen by a more rapid onset and higher magnitude of anti-hepatitis B Abs. In addition, the immunogenicity of a potent HIV gag DNA vaccine was increased in mice, as seen by higher Ab titers, a substantial reduction in the dose of
9、 DNA required to induce an Ab response, and an increase in CD8 1 T cell responses. Finally, Ab responses were enhanced by electroporation against both components of a combination HIV gag and env DNA vaccine in guinea pigs and rabbits. Therefore, cellular uptake of DNA is a signicant barrier to trans
10、fection in vivo, and electroporation appears able to overcome this barrier. The Journal of Immunology, 2000, 164: 46354640. T he prospect of inducing an immune response to a protein expressed in vivo directly from an administered DNA vaccine represents an attractive alternative to other modes of vac
11、cination. The de novo synthesis of DNA vaccine-encoded proteins mimics expression of Ags after viral infection and may improve processing and presentation to the immune system, thereby providing the advantages of live attenuated vaccines with- out the safety and stability concerns associated with th
12、e adminis- tration of infectious agents. Because of these potential advantages, considerable effort has been expended in evaluating this technol- ogy (for review, see Ref. 1). Early successes in demonstrating protective efcacy in small animal models have helped to drive the testing of DNA vaccines i
13、n larger animals, culminating in several human clinical trials. Thus far, however, in only a few cases have immune responses been demonstrated in humans (24), and the magnitude of these responses has been insubstantial. Therefore, for this technology to be effective for human vaccination, more poten
14、t forms of DNA vaccines must be identied and developed. One reason for the lack of efcacy in larger animals may be inefcient uptake of DNA by cells in situ. Hence, we sought to test whether cellular uptake of DNA was a signicant limitation to efcient transfection in vivo and subsequent induction of
15、immune responses. To this end, we used the technique of electroporation, which is widely used in vitro to effectively introduce DNA into eukaryotic cells and bacteria. Application of short electrical pulses to the target cells permeabilizes the cell membrane, thereby facil- itating DNA uptake. Recen
16、tly, it has been found that applying an electric eld to tissues in vivo signicantly increases DNA uptake and gene expression (for review, see Ref. 5). This has been shown for reporter genes and for genes of interest for therapeutic appli- cations, such as erythropoietin (6) and HSV-TK (7). Among the tissues targeted for in vivo electroporation have been skin (8, 9), liver (10, 11), tumors (1214), and muscle (15). Facilitation of gene expression in vivo by electrop
