Plasmid DNA-recombinant Opc protein complexes for nasal DNA immunization.
The nasal mucosa may provide a simple, non-invasive route to deliver DNA encoding genes that stimulate a specific immune response. Based on this, a new approach using pCMVbeta-gal plasmid DNA complexed to the Opc meningococcal outer membrane protein was assayed for. Optimal conditions of interaction were established between recombinant Opc protein and pCMVbeta-gal plasmid DNA. Complexes were fully characterized by electrophoresis analysis, DNAse resistance assay and transmission electron microscopy. DNA-protein complexes were also evaluated in in vitro transfection experiments. After the characterisation of complexes, Balb/c mice were intranasal (i.n.) and intramuscularly (i.m.) immunized. The humoral immune response against beta-galactosidase was measured by ELISA. The proliferative response in the spleen lymph nodes was also measured. Complexes administered by i.n. route induced both systemic and mucosal antibody responses. This behavior was not observed with the naked DNA. Finally, a lymphoproliferative response specific to beta-galactosidase induced by DNA-protein complexes was also detected.
Musacchio
A
A
Division of Vaccines, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, C.P. 10600, C. Habana, Cuba. alexis.musacchio@cigb.edu.cu
Quintana
D
D
Herrera
A M
AM
Sandez
B
B
Alvarez
J C
JC
Falcón
V
V
la Rosa
M C
MC
Alvarez
F
F
Pichardo
D
D
D000281:Administration, Intranasal; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D019556:COS Cells; D002460:Cell Line; D007115:Immunization Schedule; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure; D014162:Transfection; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D019444:Vaccines, DNA; C074517:opc protein, bacteria