Abstract
Purpose. A plasmid-based gene expression system was complexed with protective, interactive, and non-condensing (PINC™) polymer system and administered with Medi-Jector™, a needle-free injection device (NFID), to achieve high and sustained levels of antigen-specific antibodies in blood circulation.
Methods. Human growth hormone (hGH) or bacterial β-galactosidase gene expression plasmids driven by a cytomegalovirus (CMV) promoter were formulated in saline or complexed with a PINC polymer, polyvinylpyrrolidone (PVP), and intramuscularly or subcutaneously administered into dogs and pigs using a 22-gauge needle or a NFID. The hGH-specific IgG titers in serum were measured by an ELISA. β-galactosidase expression was measured in injected muscles by an enzymatic assay or immunohistochemistry. The effect of NFID on DNA stability and topology was assessed by gel electrophoresis.
Results. Intramuscular (i.m.) or subcutaneous (s.c.) injection of a hGH expression plasmid pCMV-hGH (0.05-0.5 mg/kg) in dogs and pigs elicited antigen-specific IgG antibody titers to expressed hGH. With both routes of injection, pDNA delivery by a NFID was superior to pDNA injection by needle. The magnitude of hGH-specific IgG titers with NFID was 15−20-fold higher than needle injection when pDNA was complexed with PVP, and only 3−4-fold higher with pDNA in saline. The transfection efficiency in the injected muscle, as measured by β-galaclosidase expression, following i.m. injection of pCMV-β-galaclosidase/PVP, was not significantly different between needle and NFID-injected groups.
Conclusions. These data demonstrate that the combination of pDNA/ PVP complexes and a NFID act synergistically to achieve high and sustained levels of antigen-specific IgG response to expressed antigen. This gene delivery approach may offer advantage over needle injection of naked DNA for the development of genetic vaccines.
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REFERENCES
D. Tang, M. DeVit, and S. A. Johnston. Genetic immunization is a simple method for eliciting an immune response. Nature 356:152–154 (1992).
M. A. Liu, M. R. Hilleman, and R. Kurth, (eds.). DNA vaccines: A new era in vaccinology. Ann. N.Y. Acad. Sci. 772 (1995).
H. L. Davis. Plasmid DNA expression systems for the purpose of immunization. Curr. Opinion Biotechnol. 8:635–646 (1997).
J. P. Lewis, G. J. M. Cox, van-Drunen-Little-van-den-Hurks-S., and L. A. Babiuk. Polynucleotide vaccines in animals: enhancing and modulating responses. Vaccine 15:861–864 (1997).
G. J. Cox, T. J. Zamb, and L. A. Babiuk. Bovine herpes virus 1: immune response in mice and cattle injected with plasmid DNA. J. Virol. 67:5664–5667 (1993).
J. B. Ulmer, J. C. Sadoff, and M. A. Liu. DNA vaccines. Cur. Opin. Immunol. 8:531–536 (1993).
J. B. Ulmer, J. J. Donnelly, S. E. Parker, G. H. Rohdes, P. L. Felgner, V. J. Dwarki, S. H. Gromkowski, R. R. Deck, C. M. DeWitt, A. Friedman, L. A. Hawel, K. R. Leander, D. Martinez, H. C. Perry, J. W. Shiver, D. L. Montgomery, and M. A. Liu. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 259:1745–1749 (1993).
B. Wang, K. E. Ugen, V. Srikantan, M. G. Agadjanyan, K. Dang, Y. Rafaeli, A. L. Sato, J. Boyer, W. V. Williams, and B. Weiner. Gene inoculation generates immune responses against human immunodeficiency virus type 1. Proc. Natl. Acad. Sci. U.S.A. 90:4156–4160 (1993).
H. Davis, M. L. Michel, M. Mancini, M. Schleef, and R. G. Wahlen. Direct gene transfer in skeletal muscle: plasmid DNA-based immunization against the hepatitis B virus surface antigen. Vaccine 12:1503–1509 (1994).
H. L. Davis, M. L. Michel, and R. G. Wahlen. DNA-based immunization induces continuous secretion of hepatitis B surface antigen and high levels of circulating antibody. Hum. Mol. Genet. 2:1847–1851 (1995).
L. A. Babiuk, Van-Drunen-Little, Van-den-Hurk, S. K. Tikoo, P. J. Lewis, and X. Liang. Non-viral vaccines for livestock. Vet. Immunol. Immunopathol. 54:355–363 (1996).
S. Lu, J. Arthos, D. C. Montefiori, Y. Yasutomi, K. Manson, F. Mustafa, E. Johnson, J. C. Santoro, J. Wissink, J. I. Millins, J. R. Haynes, N. L. Letvin, M. Wyand, and H. L. Robinson. Simian immunodeficiency virus DNA vaccine trail in macaques. J. Virol. 70: 3978–3991 (1996).
D. H. Fuller, M. M. Corb, S. Barnett, K. Steimer, and J. R. Haynes. Enhancement of immunodeficiency virus-specific immune responses in DNA-immunized rhesus macaques. Vaccine 15:924–926 (1997).
E. F. Fynan, R. G. Webster, D. H. Fuller, J. R. Haynes, J. C. Santoro, and H. L. Robinson. DNA vaccines: protective immunization by parenteral, mucosal, and gene-gun inoculations. Proc. Natl. Acad. Sci. U.S.A. 90:11478–11482 (1993).
H. L. Robinson, L. A. Hunt, and R. G. Webster. Protection against a lethal influenza virus challenge by immunization with a haemagglutinin-expressing plasmid DNA. Vaccine 11:957–960 (1993).
R. G. Webster, E. F. Fynan, J. C. Santoro, and H. Robinson. Protection of ferrets against influenza challenge with a DNA vaccine to the haemagglutinin. Vaccine 12:1495–1498 (1994).
Z. O. Xiang, S. Spitalink, M. Tran, W. H. Wunner, J. Cheng, and H. C. J. ERTL. Vaccination with a plasmid vector carrying the rabies virus glycoprotein gene induces protective immunity against rabies virus. Virology 199:132–140 (1994).
L. H. Davis, R. G. Wahlen, and B. A. Demeneix. Direct gene transfer into skeletal muscle in vivo: factors affecting efficiency of transfer and stability of expression. Hum. Gene Ther. 4: 151–159 (1993).
S. Jiao, P. Williams, R. K. Berg, B. A. Hodgeman, L. Liu, G. Repetto, and J. A. Wolff. Direct gene transfer into nonhuman primate myofibers in vivo. Hum. Gene Ther. 3:21–33 (1992).
P. A. Furth, A. Shamay, R. J. Wall, and L. Hennighausen. Gene transfer into somatic tissues by jet injection. Anal. Biochem. 205:365–368 (1992).
H. L. Vahlsing, M. A. Yankauckas, M. Sawdey, S. H. Gromkowski, and M. Manthrope. Immunization with plasmid DNA using a pneumatic gun. J. Immunol. Methods 175:11–22 (1994).
P. A. Furth, A. Shamay, and L. Hennighausen. Gene Transfer into mammalian cell by jet injection. Hybridoma 14:149–152 (1995).
M. Jenkins, D. Kerr, R. Fayer, and R. Wall. Serum and clostrum antibody responses induced by jet-injection of sheep with DNA encoding a Cryptosporidum parvum antigen. Vaccine 13: 1658–1664 (1995).
K. Anwer, M. Shi, M. F. French, S. R. Muller, W. Chen, Q. Liu, B. L. Proctor, J. Wang, R. J. Mumper, A. Singhal, A. P. Rolland, and H. W. Alila. Systemic effect of human growth hormone after intramuscular injection of a single dose of a muscle-specific gene medicine. Hum. Gene. Ther. 9:659–670 (1998).
R. J. Mumper, J. G. Duguid, K. Anwer, M. K. Barron, H. Nitta, and A. P. Rolland. Polyvinyl derivatives as novel interactive polymers for controlled gene delivery to muscle. Pharm. Res. 13:701–709 (1996).
R. J. Mumper, J. Wang, S. L. Klakamp, H. Nitta, K. Anwer, F. Tagliaferri, and A. P. Rolland. Protective, interactive, noncondensing (PINC) polymers for enhanced plasmid distribution and expression in rat skeletal muscle. J. Contr. Rel. 52:191–203 (1998).
D. E. Kerrr, P. A. Furth, A. M. Powell, and R. J. Wall. Expression of gene-gun injected plasmid DNA in the ovine mammary gland and in lymph nodes draining the injection site. Animal Biotech. 7:33–45 (1996).
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Anwer, K., Earle, K.A., Shi, M. et al. Synergistic Effect of Formulated Plasmid and Needle-Free Injection for Genetic Vaccines. Pharm Res 16, 889–895 (1999). https://doi.org/10.1023/A:1018834305079
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DOI: https://doi.org/10.1023/A:1018834305079