Assembly of human immunodeficiency virus (HIV) antigens on bacteriophage T4: a novel in vitro approach to construct multicomponent HIV vaccines.
Bacteriophage T4 capsid is an elongated icosahedron decorated with 155 copies of Hoc, a nonessential highly antigenic outer capsid protein. One Hoc monomer is present in the center of each major capsid protein (gp23*) hexon. We describe an in vitro assembly system which allows display of HIV antigens, p24-gag, Nef, and an engineered gp41 C-peptide trimer, on phage T4 capsid surface through Hoc-capsid interactions. In-frame fusions were constructed by splicing the human immunodeficiency virus (HIV) genes to the 5' or 3' end of the Hoc gene. The Hoc fusion proteins were expressed, purified, and displayed on hoc(-) phage particles in a defined in vitro system. Single or multiple antigens were efficiently displayed, leading to saturation of all available capsid binding sites. The displayed p24 was highly immunogenic in mice in the absence of any external adjuvant, eliciting strong p24-specific antibodies, as well as Th1 and Th2 cellular responses with a bias toward the Th2 response. The phage T4 system offers new direction and insights for HIV vaccine development with the potential to increase the breadth of both cellular and humoral immune responses.
Sathaliyawala
Taheri
T
Department of Biology, The Catholic University of America, 620 Michigan Avenue NE, Washington, DC 20064, USA.
Rao
Mangala
M
Maclean
Danielle M
DM
Birx
Deborah L
DL
Alving
Carl R
CR
Rao
Venigalla B
VB
D016915:AIDS Vaccines; D000818:Animals; D017122:Bacteriophage T4 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002213:Capsid / Q000276:immunology / Q000378:metabolism*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015488:HIV Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016655:HIV Core Protein p24 / Q000276:immunology*; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014614:Vaccines, Synthetic / Q000276:immunology
D016915:AIDS Vaccines; D036022:Capsid Proteins; D015488:HIV Antigens; D016655:HIV Core Protein p24; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic