title: Multimeric soluble CD40 ligand and GITR ligand as adjuvants for human immunodeficiency virus DNA vaccines.
abstract: For use in humans, human immunodeficiency virus (HIV) DNA vaccines may need to include immunostimulatory adjuvant molecules. CD40 ligand (CD40L), a member of the tumor necrosis factor (TNF) superfamily (TNFSF), is one candidate adjuvant, but it has been difficult to use because it is normally expressed as a trimeric membrane molecule. Soluble trimeric forms of CD40L have been produced, but in vitro data indicate that multimeric, many-trimer forms of soluble CD40L are more active. This multimerization requirement was evaluated in mice using plasmids that encoded either 1-trimer, 2-trimer, or 4-trimer soluble forms of CD40L. Fusion with the body of Acrp30 was used to produce the 2-trimer form, and fusion with the body of surfactant protein D was used to produce the 4-trimer form. Using plasmids for secreted HIV-1 antigens Gag and Env, soluble CD40L was active as an adjuvant in direct proportion to the valence of the trimers (1 < 2 < 4). These CD40L-augmented DNA vaccines elicited strong CD8(+) T-cell responses but did not elicit significant CD4(+) T-cell or antibody responses. To test the applicability of the multimeric fusion protein approach to other TNFSFs, a 4-trimer construct for the ligand of glucocorticoid-induced TNF family-related receptor (GITR) was also prepared. Multimeric soluble GITR ligand (GITRL) augmented the CD8(+) T-cell, CD4(+) T-cell, and antibody responses to DNA vaccination. In summary, multimeric CD40L and GITRL are new adjuvants for DNA vaccines. Plasmids for expressing multimeric TNFSF fusion proteins permit the rapid testing of TNFSF molecules in vivo.
authors.affiliation: Department of Medicine-0679, Stein Clinical Sciences Bldg., Room 304, University of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0679, USA. rkornbluth@ucsd.edu
authors.forename: Geoffrey W
authors.initials: GW
authors.lastname: Stone
authors.forename: Suzanne
authors.initials: S
authors.lastname: Barzee
authors.forename: Victoria
authors.initials: V
authors.lastname: Snarsky
authors.forename: Kristin
authors.initials: K
authors.lastname: Kee
authors.forename: Celsa A
authors.initials: CA
authors.lastname: Spina
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Yu
authors.forename: Richard S
authors.initials: RS
authors.lastname: Kornbluth
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D052242:Adiponectin / Q000235:genetics; D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D023201:CD40 Ligand / Q000235:genetics / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015686:Gene Products, env / Q000235:genetics / Q000276:immunology; D015683:Gene Products, gag / Q000235:genetics / Q000276:immunology; D015483:HIV Antibodies / Q000097:blood; D015497:HIV-1 / Q000276:immunology*; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000008:administration & dosage; D051379:Mice; D010957:Plasmids; D037663:Pulmonary Surfactant-Associated Protein D / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D048069:Tumor Necrosis Factors / Q000235:genetics / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D052242:Adiponectin; C499919:Adipoq protein, mouse; D000276:Adjuvants, Immunologic; D015686:Gene Products, env; D015683:Gene Products, gag; D015483:HIV Antibodies; D037663:Pulmonary Surfactant-Associated Protein D; D011993:Recombinant Fusion Proteins; C478966:Tnfsf18 protein, mouse; D048069:Tumor Necrosis Factors; D019444:Vaccines, DNA; D023201:CD40 Ligand; D007371:Interferon-gamma