title: Transgene optimization, immunogenicity and in vitro efficacy of viral vectored vaccines expressing two alleles of Plasmodium falciparum AMA1.
abstract: BACKGROUND
  Apical membrane antigen 1 (AMA1) is a leading candidate vaccine antigen against blood-stage malaria, although to date numerous clinical trials using mainly protein-in-adjuvant vaccines have shown limited success. Here we describe the pre-clinical development and optimization of recombinant human and simian adenoviral (AdHu5 and ChAd63) and orthopoxviral (MVA) vectors encoding transgene inserts for Plasmodium falciparum AMA1 (PfAMA1).
  
  
  RESULTS
  AdHu5-MVA prime-boost vaccination in mice and rabbits using these vectors encoding the 3D7 allele of PfAMA1 induced cellular immune responses as well as high-titer antibodies that showed growth inhibitory activity (GIA) against the homologous but not heterologous parasite strains. In an effort to overcome the issues of PfAMA1 antigenic polymorphism and pre-existing immunity to AdHu5, a simian adenoviral (ChAd63) vector and MVA encoding two alleles of PfAMA1 were developed. This antigen, composed of the 3D7 and FVO alleles of PfAMA1 fused in tandem and with expression driven by a single promoter, was optimized for antigen secretion and transmembrane expression. These bi-allelic PfAMA1 vaccines, when administered to mice and rabbits, demonstrated comparable immunogenicity to the mono-allelic vaccines and purified serum IgG now showed GIA against the two divergent strains of P. falciparum encoded in the vaccine. CD8(+) and CD4(+) T cell responses against epitopes that were both common and unique to the two alleles of PfAMA1 were also measured in mice.
  
  
  CONCLUSIONS
  Optimized transgene inserts encoding two divergent alleles of the same antigen can be successfully inserted into adeno- and pox-viral vaccine vectors. Adenovirus-MVA immunization leads to the induction of T cell responses common to both alleles, as well as functional antibody responses that are effective against both of the encoded strains of P. falciparum in vitro. These data support the further clinical development of these vaccine candidates in Phase I/IIa clinical trials.
authors.affiliation: The Jenner Institute, University of Oxford, Oxford, Oxfordshire, United Kingdom. sumi.biswas@ndm.ox.ac.uk
authors.forename: Sumi
authors.initials: S
authors.lastname: Biswas
authors.forename: Matthew D J
authors.initials: MD
authors.lastname: Dicks
authors.forename: Carole A
authors.initials: CA
authors.lastname: Long
authors.forename: Edmond J
authors.initials: EJ
authors.lastname: Remarque
authors.forename: Loredana
authors.initials: L
authors.lastname: Siani
authors.forename: Stefano
authors.initials: S
authors.lastname: Colloca
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Cottingham
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Holder
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Gilbert
authors.forename: Adrian V S
authors.initials: AV
authors.lastname: Hill
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Draper
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000483:Alleles; D000818:Animals; D005822:Genetic Vectors; D017780:Malaria Vaccines / Q000235:genetics / Q000276:immunology*; D016778:Malaria, Falciparum / Q000517:prevention & control*; D051379:Mice; D018155:Orthopoxvirus / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics* / Q000276:immunology; D011817:Rabbits; D019076:Transgenes
chemical_list: D017780:Malaria Vaccines