AEGiS-11IAC: Chimeric human immunodeficiency virus gag particles as an antigen presenting vehicle.

11th International AIDS Conference


Vancouver, British Columbia — July 7-12, 1996

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Chimeric human immunodeficiency virus gag particles as an antigen presenting vehicle.

Int Conf AIDS 1996 Jul 7-12; 11:7 (abstract no. Mo.A.154)
Nariyoshi H, Yasuda A, Kurata T, Kojima A; Dept. of Pathology, NIH, Tokyo, Japan. Fax: 3-5285-1189. E-mail: nhoshika@nih.go.jp.

OBJECTIVE: Viral antigens usually show the highest immunogenicity when they form virus-like particle structures. In this study, we examined whether human immunodeficiency virus (HIV) Gag particles work as immunogenic carriers for a hepatitis C virus (HCV) core epitope.

METHODS: The HIV-1 gag gene (BH10) was deleted of the p2 region by introduction of Kpn-I sites at its 5' - and 3' -ends and then Kpn-I digestion. Synthetic oligonucleotides encoding the N-terminal 28 amino acids of HCV core (C28) were inserted at the Kpn-I site to create hybrid gag-C28 genes. The hybrid genes were expressed by recombinant vaccinia virus (RVV). Expression and release of hybrid proteins were analyzed by immunoblotting. Particle formation was examined by transmission and scanning electron microscopy of RVV-infected cells. Antibody responses in RVV-infected mice were determined by ELISA.

RESULTS: RVV-infected cells expressed Gag proteins with the increased molecular mass when analyzed with anti-p17 and anti-p24 monoclonal antibodies. The larger Gag proteins also reacted specifically to sera from HCV-infected patients and from rabbits immunized with recombinant HCV core. Gag-like particle formations prepared from culture medium by ultracentrifugation contained HCV-core-specific and Gag-specific proteins. Extracellular Gag-like particles were observed by electron microscopy of RVV-infected cells. Mice inoculated with RVV induced anti-C28 as well as anti-p17 antibody responses.

CONCLUSIONS: We showed that a Gag-C28 mosaic protein assembles into recombinant Gag-like particles and that Gag-C28 particles process marked immunogenicity in mice. These results suggest that HIV Gag particles can be useful for carrying immunogenic epitopes and vaccine vehicles.

Keywords: AEGIS, HIV, Gene Products, gag, Genes, gag, HIV-1, HIV Antibodies, Vaccinia virus, Immunoblotting, Chimeric Proteins, Epitopes, Human, Animal, Mice, Rabbits, genetics, ICA11KWDaegis,hiv,geneproducts,gag,genes,gag,hiv-1,hivantibodies,vacciniavirus,immunoblotting,chimericproteins,epitopes,human,animal,mice,rabbits,genetics,ica11

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MoA154

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