Construction, Characterization, and Immunogenicity of a Multigene Modified Vaccinia Ankara (MVA) Vaccine Based on HIV Type 1 Subtype C

Candidate vaccines composed of a DNA construct to prime the immune system, followed by modified vaccinia Ankara (MVA) containing matching genes as a booster vaccination, have produced encouraging immune responses in human volunteers. This study presents the detailed construction and characterization of a recombinant MVA that will be tested in combination with a DNA vaccine in Phase I clinical trials in South Africa and the United States. To match recently transmitted viruses in the southern African region and to maximize epitope coverage, the vaccines were constructed to contain five HIV-1 subtype C genes, namely gag, reverse transcriptase, tat, and nef (grttn), expressed as a polyprotein, and a truncated env (gp150). An initial recombinant MVA construct containing wild-type env was found to be genetically unstable, and thus a human codon-optimized gene was used. Grttn and gp150 were inserted into two different sites in MVA yielding a double recombinant, SAAVI MVA-C. The recombinant MVA was shown to be genetically stable and high level expression of the transgenes was observed. Env retained infectivity in a functional infectivity assay despite a point mutation that arose during virus generation. Mice inoculated with SAAVI MVA-C at various doses developed high levels of Gag, RT, and Env-specific CD8(+) and CD4(+) T cells, and some of these responses could be boosted by a second inoculation. An accompanying paper describes the immunogenicity of SAAVI MVA-C when given in combination with SAAVI DNA-C.