Synergistic effect of formulated plasmid and needle-free injection for genetic vaccines.
OBJECTIVE
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 beta-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. Beta-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 beta-galactosidase expression, following i.m. injection of pCMV-betagalactosidase/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.
GeneMedicine, Inc., The Woodlands, Texas 77381-4248, USA. anwerk@genemedicine.com
K
K
Anwer
K A
KA
Earle
M
M
Shi
J
J
Wang
R J
RJ
Mumper
B
B
Proctor
K
K
Jansa
H C
HC
Ledebur
S
S
Davis
W
W
Eaglstein
A P
AP
Rolland
D000818:Animals; D004285:Dogs; D005260:Female; D005822:Genetic Vectors / Q000008:administration & dosage; D019382:Human Growth Hormone / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007277:Injections, Jet; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000008:administration & dosage; D011205:Povidone / Q000008:administration & dosage / Q000737:chemistry; D013552:Swine; D014611:Vaccination / Q000379:methods*; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology; D001616:beta-Galactosidase / Q000096:biosynthesis
D007074:Immunoglobulin G; D011108:Polymers; D019444:Vaccines, DNA; D019382:Human Growth Hormone; D001616:beta-Galactosidase; D011205:Povidone