Selected strategies to augment polynucleotide immunization.
We sought to amplify the immune response to polynucleotide immunization through co-delivery of complementary DNA (cDNA) encoding a cytokine or co-stimulatory molecule to enhance antigen presentation. In the context of intramuscular immunization, we examined co-delivery of cDNAs for B7-1 and human carcinoembryonic antigen (CEA) within separate plasmids or a dual plasmid with two independent expression cassettes. Intramuscular delivery of the dual expression plasmid produced anti-CEA antibody responses and antitumor effects superior to those generated by plasmid DNA encoding CEA alone. However, co-delivery of cDNAs encoding B7-1 and CEA in the form of two separate plasmids produced no augmentation. The importance of single plasmid delivery suggests the effectiveness of this strategy is contingent upon co-expression of B7-1 and CEA within the same cell. The success of cutaneous polynucleotide immunization by particle bombardment is thought to derive largely from the presence of Langerhans cells within the skin. We hypothesized that co-delivery of plasmid DNA encoding granulocyte-macrophage colony stimulating factor (GM-CSF) by particle bombardment would enhance the antigen presenting capacity of Langerhans cells at the inoculation site similar to its effects in vitro. Augmentation of CEA-specific lymphoblastic transformation and antibody response was observed when plasmid GM-CSF (pGM-CSF) was administered 3 days prior to each dose of plasmid DNA encoding CEA. These strategies for augmentation of immune response to polynucleotide immunization should be applicable to a wide variety of antigenic targets including infectious agents and other tumor-associated antigens.
Conry
R M
RM
Gene Therapy Program, Department of Medicine, University of Alabama at Birmingham, USA.
Widera
G
G
LoBuglio
A F
AF
Fuller
J T
JT
Moore
S E
SE
Barlow
D L
DL
Turner
J
J
Yang
N S
NS
Curiel
D T
DT
D000818:Animals; D018122:B7-1 Antigen / Q000235:genetics* / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000235:genetics* / Q000276:immunology; D005260:Female; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D007114:Immunization; D051379:Mice; D008810:Mice, Inbred C57BL; D011119:Polynucleotides / Q000276:immunology; D014407:Tumor Cells, Cultured
D018122:B7-1 Antigen; D002272:Carcinoembryonic Antigen; D011119:Polynucleotides; D016178:Granulocyte-Macrophage Colony-Stimulating Factor