Cationic submicron emulsions for pulmonary DNA immunization.
Pulmonary immunization against inhaled pathogens such as Mycobacterium tuberculosis would induce local and systemic immune responses and protect from entry and dissemination of the pathogen. The aim of this study was to evaluate cationic submicron emulsion as a potential carrier for DNA vaccines to the lung. DNA loaded emulsions were 128-152 nm in size and retained positive zeta potential above +40 mV during 3 months of storage. Loading efficiency was above 99%, DNA was protected from DNase I degradation up to 60 min and was stable in presence of 75% fetal calf serum (FCS). The plasmid DNA was detected in the endo-lysosomal compartment of the human bronchial cell line, Calu-3, 6 h after application. No cytotoxic effect on these cells was observed. Human dendritic cells were matured in presence of DNA loaded emulsion, although to a lesser extent than DNA solution indicating slower release and lower exposure to unmethylated CpG sequences. These results indicate that cationic submicron emulsions are potential DNA vaccine carriers to the lung since they are able to transfect pulmonary epithelial cells, which possibly induce cross priming of antigen presenting cells and directly activate dendritic cells, resulting in stimulation of antigen specific T-cells.
Bivas-Benita
Maytal
M
Leiden/Amsterdam Center for Drug Research, Division of Pharmaceutical Technology, P.O. Box 9502, 2300 RA Leiden, The Netherlands. m.bivas@lacdr.leidenuniv.nl
Oudshoorn
Marion
M
Romeijn
Stefan
S
van Meijgaarden
Krista
K
Koerten
Henk
H
van der Meulen
Hans
H
Lambert
Gregory
G
Ottenhoff
Tom
T
Benita
Simon
S
Junginger
Hans
H
Borchard
Gerrit
G
D000217:Acyltransferases / Q000235:genetics* / Q000276:immunology; D000327:Adsorption; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D003713:Dendritic Cells / Q000502:physiology; D016503:Drug Delivery Systems; D004655:Emulsions; D006801:Humans; D007114:Immunization; D010316:Particle Size; D032581:Tuberculosis Vaccines / Q000008:administration & dosage*; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004655:Emulsions; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA; D000217:Acyltransferases; C121433:antigen 85B, Mycobacterium tuberculosis