Plasmids encoding granulocyte-macrophage colony-stimulating factor and CD154 enhance the immune response to genetic vaccines

We examined whether plasmids encoding granulocyte-macrophage colony-stimula ting factor (pGM-CSF) or CD40-ligand (pCD40L) could modify the immune respo nse to antigen encoded by co-injected plasmid DNA. For this we used as anti gen Escherichia coli beta galactosidase (beta -gal), encoded by the plasmid pLacZ. We found that intradermal co-injection of pLacZ with both pGM-CSF a nd pCD40L enhanced the anti-beta -gal IgG response by approximately two ord ers of magnitude compared to injections of pLacZ alone. Co-injection of bot h pGM-CSF and pCD40L with pLacZ significantly enhanced antigen-specific IgG , and in particular IgG(2a), over that of animals co-injected with pLacZ an d either pGM-CSF or pCD40L. We found that co-injection of pGM-CSF and pCD40 L with pLacZ enhanced the generation of beta -gal-specific cytotoxic T cell s, and allowed for a significant expansion of cD8(+) T cells from splenocyt es co-cultured with beta -gal expressing stimulator cells. The immunostimul atory effects induced by pGM-CSF or pCD40L required injection of these plas mids to the same site that received pLacZ. 'Priming experiments, where the site of injection was pre-injected with either plasmid adjuvant, showed tha t pGM-CSF, but not pCD40L, could enhance the anti-beta -gal immune response induced by subsequently administered plasmid antigen. We conclude that pla smids encoding GM-CSF and CD154 are particularly effective genetic adjuvant s when used together to enhance the humoral and cellular immune response to a plasmid-encoded antigen. (C) 2001 Elsevier Science Ltd. All rights reser ved.