Summary
Immunisation with purified DNA is a powerful technique for inducing immune responses. The concept is very simple, involving insertion of the gene encoding the antigen of choice into a bacterial plasmid, and injection of the plasmid into the host where the antigen is expressed and induces humoral and cellular immunity. This technology can induce immunity to all antigens that can be encoded by DNA; this includes all protein, but not carbohydrate, antigens. DNA immunisation appears to result in presentation of antigens to the host’s immune system in a natural form, similar to that achieved with live attenuated vaccines.
The most efficacious routes for DNA immunisation are bombardment with particles coated with DNA (gene-gun), followed by intramuscular and intradermal administration. The efficiency of transfection of host cells is low, but sufficient to induce immunological responsiveness. The DNA plasmid is retained in the transfected cells in an unintegrated form for the life of the cell. The majority of transfected cells are eliminated, but residual expression has been detected for longer periods.
In animal model systems, DNA immunisation has been shown to induce protective immunity to influenza, herpes, rabies, hepatitis B and lymphocytic choriomeningitis viruses, and to malaria and mycobacteria. However, strategies to induce protective immunity to HIV and other disease agents remain to be developed. DNA vaccines permit modulation of the immune response by altering the route or method of DNA administration, by including immunostimulatory sequences in the plasmid, and by co-administration of cytokine genes with the gene encoding the antigen of interest. A T helper 1 response provides cell-mediated immune killing of infected cells and neutralising antibody production, while a T helper 2 response induces IgE and allergic responses.
The advantages of DNA immunisation are: (i) similarity to live attenuated vaccination but without the possibility of contamination with undesirable agents; (ii) correct presentation of antigen; (iii) combinations of DNA-encoded antigens and/or cytokines may be administered; (iv) genetic stability; (v) potential speed of making new vaccines with genetic identity; (vi) development of vaccines for agents that cannot be grown in culture; (vii) no need for a cold chain; and (viii) possibility of modulation of the immune response. The perceived risks include: (i) integration of the plasmid into the host genome; (ii) induction of anti-DNA antibodies and autoimmunity; and (iii) induction of tolerance. The available information concerning safety is encouraging, with the risk of integration being considered to be orders of magnitude below the spontaneous mutation frequency in humans.
DNA immunisation offers the possibility of extending the control of infectious diseases far beyond those that are currently controlled by conventional and recombinant vaccines, to include vaccines for parasites and cancer. However, it is currently too early to predict the future extent of use of DNA vaccines in human immunisation programmes because the initial clinical trials are still in progress.
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Fauci AS. In: Report of the task force on microbiology and infectious diseases. Bethesda: U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, 1992: 1–55
Gierer A, Schramm G. The infectivity of ribonucleic acid from tobacco mosaic virus. Z Naturforsch 1956; 116: 138–42
Alexander HE, Koch G, Mountain JM, et al. Infectivity of ribonucleic acid from poliovirus in cell monolayers. J Exp Med 1958; 493–506
Racaniello VR, Baltimore D. Cloned poliovirus complementary DNA is infectious in mammalian cells. Science 1981; 214: 916–9
Fung Y-KT, Crittenden LB, Fadly AM, et al. Tumor induction by direct injection of cloned v-src DNA into chickens. Proc Natl Acad Sci USA 1983; 80: 353–7
Benvenisty N, Reshef L. Direct introduction of genes into rats and expression of the genes. Proc Natl Acad Sci USA 1986; 83: 9551–5
Atanasiu P. Production de tumeurs chez le hamster par inoculation d’acide desoxyribonucleique extrait de cultures de tissus par le virus de polyome. CR Hebd Seances Acad Sci Ser D 1962; 254: 4228–30
Israel MA, Chan HW, Hourihan SL, et al. Biological activity of polyoma viral DNA in mice and hamsters. J Virol 1979; 29: 990–6
Wolff JA, Malone RW, Williams P, et al. Direct gene transfer into mouse muscle in vivo. Science 1990; 247: 1465–8
Ulmer JB, Donnelly JJ, Parker SE, et al. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 1993; 259: 1745–9
Cox G, Zamb TJ, Babiuk LA. Bovine herpesvirus 1: immune responses in mice and cattle injected with plasmid DNA. J Virol 1993; 67: 5664–7
Davis HL, Michel M, Whalen R. DNA-based immunization induces continuous secretion of hepatitis B surface antigen and high levels of circulating antibody. Hum Mol Genet 1993; 2: 1847–51
Wang B, Ugen KE, Rikantan V, et al. Gene inoculation generates immune responses against immunodeficiency virus type 1. Proc Natl Acad Sci USA 1993; 90: 4156–60
Tang D-C, De Vit M, Johnston SA. Genetic immunization is a simple method for eliciting an immune response. Nature 1992; 356: 152
Hunt LA, Brown DW, Robinson HL, et al. Retrovirus-expressed hemagglutinin protects against lethal influenza virus infections. J Virol 1988; 62: 3014–9
Robinson HL, Hunt LA, Webster RG. Protection against a lethal influenza virus challenge by immunization with a haemagglutinin-expressing plasmid DNA. Vaccine 1993; 11: 957–60
Vahlsing HL, Yankauckas MA, Sawdey M, et al. Immunization with plasmid DNA using a pneumatic gun. J Immunol Methods 1994; 175: 11–22
Fynan EF, Webster RG, Fuller DH, et al. DNA vaccines: protective immunizations by parenteral, mucosal, and gene-gun inoculations. Proc Natl Acad Sci USA 1993; 90: 11478–82
Freeman GJ, Boriello F, Hodes RH, et al. Uncovering of a functional alternative CTL-4 counter-receptor in B7-deficient mice. Science 1993; 262: 907–8
Hatchcock KS, Lazio G, Dickler HB, et al. Identification of an alternative CTL-4 ligand costimulatory for T cell activation. Science 1993; 262: 905–7
Linsley P, Brady W, Grosmaire L, et al. Binding of the B cell activation antigen B7 to CD28 co-stimulates T cell proliferation and interleukin 2 mRNA accumulation. J Exp Med 1991; 173: 721–30
Pardoll DM, Beckerleg AM. Exposing the immunology of naked DNA vaccines. Immunity 1995; 3: 165–9
Eisenbraun MD, Fuller DH, Haynes JH. Examination of parameters affecting the elicitation of humoral immune responses by particle bombardment-mediated genetic immunization. DNA Cell Biol 1993; 12: 791
Raz E, Carson DA, Parker SE, et al. Intradermal gene immunizations: the possible role of DNA uptake in the induction of cellular immunity to viruses. Proc Natl Acad Sci USA 1994; 91: 9519–23
Hengge UR, Chan EF, Foster RA, et al. Cytokine gene expression in epidermis with biological effects following injection of naked DNA. Nature Genet 1995; 10: 161
Condon C, Watkins SC, Celluzzi CM, et al. DNA-based immunization by in vivo transfection of dendritic cells. Nat Med 1996; 2: 1122–8
Ledley FD. Nonviral gene therapy: the promise of genes as pharmaceutical products. Hum Gene Ther 1995; 6: 1129–44
Danko I, Fritz JD, Jiao S, et al. Pharmacological enhancement of in vivo foreign gene expression in muscle. Gene Ther 1994; 1: 114–21
Davis HL, Whalen RG, Demeneix BA. Direct gene transfer into skeletal muscle in vivo: factors affecting efficiency of transfer and stability of expression. Hum Gene Ther 1993; 4: 151–9
Yang N-S, Burkholder J, Roberts B, et al. In vivo and in vitro gene transfer to mammalian somatic cells by particle bombardment. Proc Natl Acad Sci USA 1990; 87: 9568–72
Williams RS, Johnston SA, Riedy M, et al. Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles. Proc Natl Acad Sci USA 1991; 88: 2726–30
Cheng L, Ziegelhoffer PR, Yang NS. In vivo promoter activity and transgene expression in mammalian somatic tissues evaluated by using particle bombardment. Proc Natl Acad Sci USA 1993; 90: 4455–9
Pertmer TM, Eisenbraun MD, McCabe D, et al. Gene-gun-based nucleic acid immunization: elicitation of humoral and cytotoxic T lymphocyte responses following epidermal delivery of nanogram quantities of DNA. Vaccine 1995; 13: 1427–30
Sizemore DR, Branstrom AA, Sadoff JC. Attenuated Shigella as a DNA delivery vehicle for DNA-mediated immunization. Science 1995; 270: 299–302
Furth PA, Shamay A, Wall RJ, et al. Gene transfer into somatic tissues by jet injection. Anal Biochem 1992; 20: 365–8
Wolff JA, Dowty ME, Jiao S, et al. Expression of naked plasmids by cultured myotubes and entry of plasmids into T tubules and caveolae of mammalian skeletal muscle. J Cell Sci 1992; 103: 1249–59
Sikes M, O’Malley Jr BW, Finegold MJ, et al. In vivo gene transfer into rabbit thyroid follicular cells by direct DNA injection. Hum Gene Ther 1994; 5: 837–44
Yovandich J, O’Malley Jr BW, Sikes M, et al. Gene transfer to synovial cells by intra-articular administration of plasmid DNA. Hum Gene Ther 1995; 6: 603–10
Coffin JM. Molecular mechanisms of nucleic acid integration. J Med Virol 1990; 31: 43–9
Manthorpe M, Cornefert-Jensen F, Hartikka J, et al. Gene therapy by intramuscular injection of plasmid DNA: studies on firefly luciferase gene expression in mice. Hum Gene Ther 1993; 4: 419–31
Nichols W, Ledwith B, Manan S, et al. Potential DNA vaccination into host cell genome. Ann NY Acad Sci 1995; 772: 30–9
Wilson IM, Grossman M, Cabrera JA, et al. A novel mechanism for achieving transgene persistence in vivo after somatic gene transfer into hepatocytes. J Biol Chem 1992; 267: 11483–9
Stankovics J, Andrews E, Crane AM, et al. Overexpression of human methylmalonyl CoA mutase in mice after in vivo gene transfer with asialo-glycoprotein/polylysine/DNA complexes. Hum Gene Ther 1994; 5: 1095–104
Wolff JA, Williams P, Acsadi G, et al. Conditions affecting direct gene transfer into rodent muscle in vivo. Biotechnology 1991; 11: 474–85
Lew D, Parker SE, Latimer T, et al. Cancer gene therapy using plasmid DNA: pharmacokinetics study of DNA following injection in mice. Hum Gene Ther 1995; 6: 553–64
Ledley TAS, Ledley FD. A multicompartment, numerical model of cellular events in the pharmacokinetics of gene therapies. Hum Gene Ther 1994; 5: 679–91
Ertl HCJ, Verma P, He Z, et al. Plasmid vectors as anti-viral vaccines. Ann NY Acad Sci 1995; 772: 77–87
Friedman HM, Yee A, Diggelmann H, et al. Use of a glucocorticoid-inducible promoter for expression of herpes simplex virus type 1 glycoprotein gC1, a cytotoxic protein in mammalian cells. Mol Cell Biol 1989; 9: 2303–14
Schorr J, Moritz P, Seddon T, et al. Plasmid DNA for human gene therapy and DNA vaccines: production and quality assurance. Ann NY Acad Sci 1995; 772: 271–3
Cheng L, Ziegelhoffer PR, Yang N-S. In vivo promoter activity and transgene expression in mammalian somatic tissues evaluated by using particle bombardment. Proc Natl Acad Sci USA 1993; 90: 4455–9
Chapman BS, Thayer RM, Vincent KA, et al. Effect of intron A from human cytomegalovirus (Towne) immediate-early gene on heterologous expression in mammalian cells. Nucleic Acids Res 1991; 19: 3979–86
Kass-Eisler A, Li K, Leinwand LA. Prospects for gene therapy with direct injection of polynucleotides. Ann NY Acad Sci 1995; 772: 232–40
Sato Y, Roman M, Tighe H, et al. Immunostimulatory DNA sequences necessary for effective intradermal gene immunization. Science 1996; 273: 352–4
Harms JS, Splitter GA. Interferon-gamma inhibits transgene expression driven by SV40 or CMV promoters but augments expression driven by the mammalian MHC I promoter. Hum Gene Ther 1995; 6: 1291–7
Montgomery DL, Shiver JW, Leander KR, et al. Heterologous and homologous protection against influenza A by DNA vaccination: optimization of DNA vectors. DNA Cell Biol 1993; 12: 777–83
Robinson HL, Hunt LA, Webster RG. Protection against a lethal influenza virus challenge by immunization with a hemagglu-tinin-expressing plasmid DNA. American Society for Virology 1992 Annual Meeting: 1992 Jul 11–15; Ithaca (NY), 14
Manickan E, Rouse RJD, Yu Z, et al. Genetic immunization against herpes simplex virus: protection is mediated by CD4+ T lymphocytes. J Immunol 1995; 155: 259–65
Xiang ZQ, Spitalnik S, Tran M, et al. Vaccination with a plasmid vector carrying the rabies virus glycoprotein gene induces protective immunity against rabies virus. Virology 1994; 199: 132–40
Lu S, Santoro JC, Fuller DH, et al. Use of DNAs expressing HIV-1 env and noninfectious HIV-1 particles to raise antibody responses in mice. Virology 1995; 209: 147–54
Martins LP, Lau LL, Asano MS. DNA vaccination against persistent viral infection. J Virol 1995; 69: 2574–82
Yokoyama M, Zhang J, Whitton JL. DNA immunization confers protection against lethal lymphocytic choriomeningitis virus infection. J Virol 1995; 69: 2684–8
Zarozinski CC, Fynan EF, Selin LK, et al. Protective CTL-dependent immunity and enhanced immunopathology in mice immunized by particle bombardment with DNA encoding an internal virion protein. J Immunol 1995; 154: 4010–7
Sedegah M, Hedstrom R, Hobart P, et al. Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein. Proc Natl Acad Sci USA 1994; 91: 9866–70
Mor G, Klinman DM, Shapiro S, et al. Complexity of the cytokine and antibody response elicited by immunizing mice with Plasmodium yoelii circumsporozoite protein plasmid DNA. J Immunol 1995; 155: 2039–46
Nabel GJ, Nabel EG, Yang ZY, et al. Direct gene transfer with DNA-liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans. Proc Natl Acad Sci USA 1993; 90: 11307–11
Murphy BR, Webster RG. Orthomyxoviruses. In: Fields BN, Knipe DM, Howley PM, et al., editors. Fields’ virology. 3rd ed. New York: Raven Press, 1996: 1397–445
Andrews ME, Coupar BEH. Efficacy of influenza haemagglutinin and nucleoprotein as protective antigens against influenza virus infection in mice. Scand J Immunol 1988; 28: 81–5
Donnelly JJ, Friedman A, Martinez D, et al. Preclinical efficacy of a prototype DNA vaccine: enhanced protection against antigenic drift in influenza virus. Nat Med 1995; 1: 583–7
Robinson HL, Boyle CA, Feltquate DM, et al. DNA immunization for influenza virus: studies using hemagglutinin and nucleoprotein-expressing DNAs. J Infect Dis 1997. In press
Justewicz DM, Morin MJ, Robinson HL, et al. Antibody-forming cell response to virus challenge in mice immunized with DNA encoding the influenza virus hemagglutinin. J Virol 1995; 69: 7712–7
Boyle CM, Morin M, Webster RG, et al. Role of different lymphoid tissues in the initiation and maintenance of DNA-raised antibody responses to the influenza virus H1 glycoprotein. J Virol 1996; 70: 9074–8
Justewicz DM, Webster RG. Long-term maintenance of B cell immunity to influenza virus hemagglutinin in mice following DNA-based immunization. Virology 1996; 224: 10–7
Webster RG, Fynan EF, Santoro JC, et al. Protection of ferrets against influenza challenge with a DNA vaccine to the haemagglutinin. Vaccine 1994; 12: 1495–8
Pisetsky DS, Reich C, Crowley SD, et al. Immunological properties of bacterial DNA. Ann NY Acad Sci 1995; 772: 152–63
Feltquate DM, Heaney S, Webster RG, et al. Different TH cell-types and antibody isotypes generated by saline and gene-gun DNA immunization. J Immunol 1997; 158: 2278–84
Fuller DH, Haynes JR. A qualitative progression in HIV type 1 glycoprotein 120-specific cytotoxic cellular and humoral immune responses in mice receiving a DNA-based glycoprotein 120 vaccine. AIDS Res Hum Retrovir 1996; 10: 1433–41
Troutt AB, Kelso A. Cytokine genes, regulation of. In: Roitt IM, Dives PJ, editors. Encyclopedia of immunology. London: Academic Press, 1992: 434–6
Fischer HG, Frosch S, Reske K, et al. Granulocyte-macrophage colony-stimulating factor activates macrophages derived from bone marrow cultures to synthesis of MHC class II molecules and to augmented antigen presentation function. J Immunol 1988; 141: 3882–8
Heufler C, Koch F, Schuler G. Granulocyte-macrophage colony-stimulating factor and interleukin 1 mediate the maturation of murine epidermal Langerhans cells into potent immunostimulatory cells. J Exp Med 1988; 167: 700–5
Xiang Z, Ertl HCJ. Manipulation of the immune response to a plasmid-encoded viral antigen by coinoculation with plasmids expressing cytokines. Immunity 1995; 2: 129–35
Steeg PS, Moore RN, Johnson HM. Regulation of murine macrophage Ia antigen expression by a lymphokine with immune interferon activity. J Exp Med 1982; 156: 1780–93
Fabry Z, Raine CS, Hart MN. Nervous tissue as an immune compartment: the dialect of the immune response in the CNS. Immunol Today 1994; 15: 218–24
Ramshaw IA, Leong KH, Ramsay AJ, et al. Novel vaccine strategies using recombinant fowlpox viruses expressing cytokine genes. Molecular Approaches to the Control of Infectious Diseases: 1994 Oct 5–9; Cold Spring Harbor (NY), 116
Ramshaw IA, Leong KH, Ramsay AJ, et al. Induction of protective immunity against influenza virus using DNA and recombinant avipoxvirus vectors. In: Brown LE, Hampson AW, Webster RG, editors. Options for the control of influenza III. Amsterdam: Elsevier Science, 1996: 772–6
Davis HL, Mancini M, Michel M-L, et al. DNA-based immunization overcomes H-2 haplotype restricted non-responsiveness to hepatitis B surface antigen in mice. Molecular Approaches to the Control of Infectious Diseases: 1994 Oct 5–9; Cold Spring Harbor (NY), 119
Guidotti LG, Ando K, Hobbs MV, et al. Cytotoxic T lymphocytes inhibit hepatitis B virus gene expression by a non-cytolytic mechanism in transgenic mice. Proc Natl Acad Sci USA 1994; 91: 3764–8
Bolognesi DP. The dilemma of developing and testing AIDS vaccines. In: Cooper BM, Temin RG, Suden B, editors. The DNA provirus: Howard Temin’s scientific legacy. Washington (DC): American Society for Microbiology, 1995: 301–12
Lu S, Arthos J, Montefiori DC, et al. Simian immunodeficiency virus DNA vaccine trial in macaques. J Virol 1996; 70: 3978–91
Robinson HL, Feltquate DM, Morin MJ. DNA vaccines: a new approach to immunization. In: Brown F, Chanock R, Ginsberg H, editors. Vaccines 95. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press, 1995: 69–75
Michel M-L, Davis HL, Schleef M, et al. DNA-mediated immunization to the hepatitis B surface antigen in mice: aspects of the humoral response mimic hepatitis B viral infection in humans. Proc Natl Acad Sci USA 1995; 92: 5307–11
Kodihalli S, Haynes JR, Robinson HL, et al. Cross protection among lethal H5N2 influenza viruses induced by DNA vaccine to the hemagglutinin. J Virol 1997; 71: 3391–6
Ada GL. Vaccines and immune responses. In: Webster RG, Granoff A, editors. Encyclopedia of virology. London: Academic Press, 1994: 1503–7
Sch¥del F, Aguado M-T, Lambert P-H. Introduction: nucleic acid vaccines. Vaccine 1994; 12: 1491–2
Robertson JS. Safety considerations for nucleic acid vaccines. Vaccine 1994; 12: 1526–8
Koffler D, Carr RI, Agnello V, et al. Antibodies to polynucleotides: distribution in human serums. Science 1969; 166: 1648–9
Parker SE, Vahlsing LH, Serfilipp LM, et al. Cancer gene therapy using plasmid DNA: safety evaluation in rodents and non-human primates. Hum Gene Ther 1995; 6: 575–90
Griffiths E. Assuring the safety and efficacy of DNA vaccines. Ann NY Acad Sci 1995; 772: 152–63
Hasty P, Rivera-Perez J, Bradley A. The length of homology required for gene targeting in embryonic stem cell. Mol Cell Biol 1991; 11: 5586–91
Wolff JA, Ludtke JJ, Acsadi G, et al. A long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum Mol Genet 1992; 1: 363–9
Temin HM. Overview of biological effects of addition of DNA molecules to cells. J Med Virol 1990; 31: 13–7
Hilleman MR. DNA vectors: precedents and safety. Ann NY Acad Sci 1995; 772: 1–14
Conference Report. First humans receive naked HIV DNA vaccine. AIDS Weekly Plus 1996; 1: 5
Braciale TJ. Naked DNA and vaccine design. Trends Microbiol 1993; 1: 323–4
Hohlfeld R, Engel AG. The immunobiology of muscle. Immunol Today 1994; 15: 269–74
Bevan MJ. Cross-priming for a secondary cytotoxic response to minor H antigens with H-2 congenic cells which do not cross-react in the cytotoxic assay. J Exp Med 1976; 143: 1283–8
Kovacsovics-Bankowski M, Rock KL. A phagosome-to-cytosol pathway for exogenous antigens presented on MHC class I molecules. Science 1995; 267: 243–6
Rouse BT, Nair S, Rouse RJD, et al. DNA vaccines and immunity to herpes simplex virus. Curr Top Microbiol Immunol 1997. In press
Coney L, Wang B, Ugen KE, et al. Facilitated DNA inoculation induces anti-HIV-1 immunity in vivo. Vaccine 1994; 12: 1545–50
Hoffman SL, Sedegah M, Hedstrom RC. Protection against malaria by immunization with a Plasmodium yoelii circumsporozoite protein nucleic acid vaccine. Vaccine 1994; 12: 1529–33
Lowrie DB, Tascon RE, Colston MJ, et al. Towards a DNA vaccine against tuberculosis. Vaccine 1994; 12: 1537–40
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Webster, R.G., Robinson, H.L. DNA Vaccines. BioDrugs 8, 273–292 (1997). https://doi.org/10.2165/00063030-199708040-00004
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DOI: https://doi.org/10.2165/00063030-199708040-00004