Multiprotein HIV type 1 clade B DNA and MVA vaccines: construction, expression, and immunogenicity in rodents of the MVA component.
Recombinant modified vaccinia virus Ankara (MVA) expressing SIV or SHIV Gag-Pol and Env, alone or in conjunction with a related DNA vaccine, effectively controls immunodeficiency virus infections in nonhuman primates. Here we describe the construction, characterization, and immunogenicity of MVA/HIV 48, a candidate HIV-1 clade B Gag-Pol-Env vaccine. A novel transfer vector was designed to allow the incorporation of HIV genes regulated by vaccinia virus promoters together with a reporter gene into a single site in the MVA genome and to automatically delete the reporter after the initial isolation of the recombinant MVA. MVA/HIV 48 contains chimeric HIV-1 HXB-2/BH10 gag-pol sequences, a deletion of integrase, inactivating point mutations in reverse transcriptase, and HIV-1 ADA env sequences with a truncation of most of the cytoplasmic domain to enhance expression on the plasma membrane. Cells infected with MVA/HIV 48 expressed HIV proteins, which were processed to the expected size. The Env was inserted into the plasma membrane and was functional in a CCR5 coreceptor-dependent cell fusion assay. Moreover, virus-like particles were released into the medium and budding particles containing Env were visualized by immunoelectron microscopy. Rodents that were immunized with MVA/HIV 48 produced antibodies, which neutralized a heterologous HIV-MN strain, and Gag-specific CD8 T cells. In the accompanying paper, we show that MVA/HIV 48 provided efficient boosting of an HIV DNA vaccine.
Wyatt
Linda S
LS
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Earl
Patricia L
PL
Liu
Jin Yan
JY
Smith
James M
JM
Montefiori
David C
DC
Robinson
Harriet L
HL
Moss
Bernard
B
D016915:AIDS Vaccines / Q000235:genetics / Q000276:immunology*; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D017353:Gene Deletion; D017930:Genes, Reporter; D005814:Genes, Viral; D015752:Genes, env; D015750:Genes, gag; D015751:Genes, pol; D006168:Guinea Pigs; D006678:HIV / Q000235:genetics; D015483:HIV Antibodies / Q000097:blood; D019426:Integrases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D015302:Simian Immunodeficiency Virus / Q000235:genetics; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology*
D016915:AIDS Vaccines; D015483:HIV Antibodies; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic; D019426:Integrases; D012194:RNA-Directed DNA Polymerase