This grant is for translational development of a scalable, nucleic acid-based formulation of cytomegalovirus-vectored vaccines that can be distributed without a cold chain. HIV candidate vaccines that use cytomegalovirus (CMV) as delivery vector and immunomodulatory adjuvant have shown extraordinary promise. In our preliminary work, for example, a rhesus cytomegalovirus-vectored SIV vaccine lacking the viral IL-10 gene (RhCMVdIL10-SIVgag) protected 4/6 infants from SIV infection. Tendel Therapies Inc. is licensing a portfolio of technology centered on these second-generation, CMV-vectored vaccines. Manufacturing and distribution of CMV-based vaccines present daunting challenges: (i) replication of CMV in culture is markedly slower than that of other vaccine vectors; (ii) CMV undergoes rapid genetic change when amplified in culture; (iii) the virus is enveloped and thus difficult to separate from cell- and virus-derived lipid bilayers of a similar size; (iv) methods for extreme concentration of the heterogenous particles are unknown; and (v) a cold chain is required for distribution. To eliminate these problems, Tendel is also licensing technology for vaccine delivery using purified CMV genomes propagated in E. coli. The technology permits efficient rescue of the genomes after introduction to mammalian cells. Similarly efficient rescue in vivo should lead to immune responses that are equivalent to those provoked by conventional vaccination with virions. We hypothesize that viral IL-10-deficient, CMV-vectored vaccine genomes (DNA) provoke immune responses that are indistinguishable from the protective responses stimulated by encapsidated live vaccine. Aim 1. Assess antigen expression and vaccine vector replication after delivery of vaccine genomes to macaques. CMV-vectored vaccines given as virions first replicate locally, leading to inflammatory cell influx, and then systemically, leading to viral gene expression in distant tissues. Our hypothesis predicts that successful rescue of vaccine genomes should lead to the same events. Aim 2. Test if innate and adaptive immune responses to vaccination with nucleic acid are comparable to protective anti-SIV responses observed previously. Previous studies have shown that protection against SIV is associated with specific immune responses, particularly Mamu-E-restricted CD8+ T cell responses. These innovative Phase I experiments will be sufficient to establish both the technical merit andin light of the proven commercial interest in CMV-vectored vaccinesthe commercial potential of Tendel's approach. Phase II experiments will build on this work to (i) demonstrate that macaques vaccinated with nucleic acid are protected against SIV challenge (ii) continue development of replication-defective HCMV-HIV Gag and Env vaccines.
Public Health Relevance Statement: We are seeking an effective vaccine against the human immunodeficiency virus (HIV), which has killed about 35 million people. The most effective current vaccine candidate is carried by another virus, cytomegalovirus, which has undesirable properties for translational development. We will test if new formulations of these vaccines as nucleic acid (DNA) stimulate strong antiviral immune responses, making this new form of the vaccine a likely tool for fighting the HIV epidemic worldwide.
Project Terms: adaptive immune response; Adjuvant; Antigens; Antiviral Agents; Attenuated Vaccines; Bacterial Artificial Chromosomes; base; Blood; Capital; Cations; CD8-Positive T-Lymphocytes; Cells; Characteristics; clinical development; Cold Chains; Cytomegalovirus; Cytomegalovirus Vaccines; design; Development; Distant; DNA; DNA Vaccines; Endotoxins; Epidemic; Escherichia coli; Evaluation; Event; Excision; experimental study; fighting; Formulation; Gene Expression; Generations; Genome; global health; Goals; Government; Grant; Hand; Harvest; Heterogeneity; Histology; HIV; HIV vaccine; Human; Human Resources; IL10 gene; Immune; Immune response; Immunize; immunoregulation; improved; in vivo; Infant; Infection; Inflammatory; Injections; innovation; interest; Interleukin-10; Lead; Left; Licensing; Light; Lipid Bilayers; Macaca; Macaca mulatta; Mammalian Cell; Methods; Mutation; Nucleic Acids; particle; Peripheral Blood Mononuclear Cell; Persons; Phase; Phenotype; Polymers; Production; Property; recombinase; response; restriction enzyme; Rhesus; Site; SIV; SIV Vaccines; T cell response; Technology; Testing; Time; tissue/cell culture; Tissues; tool; transcriptomics; Transfection; Vaccinated; Vaccination; Vaccine Antigen; vaccine candidate; vaccine delivery; vaccine development; Vaccines; vector; vector vaccine; Viral; Viral Genes; Virion; Virus; Work
