Alfonso S Gozalo

Protection of Aotus Monkeys by Plasmodium falciparum EBA-175 Region II DNA Prime—Protein Boost Immunization Regimen

Aotus monkeys received 4 doses of Plasmodium falciparum EBA-175 region II vaccine as plasmid DNA (Dv-Dv) or recombinant protein in adjuvant (Pv-Pv) or as 3 doses of DNA and 1 dose of protein (Dv-Pv). After 3 doses, antibody titers were approximately 10(4) in DNA-immunized monkeys and 10(6) in protein-immunized monkeys. A fourth dose did not significantly boost antibody responses in the Dv-Dv only or Pv-Pv only groups, but titers were boosted to approximately 10(6) in monkeys in the Dv-Pv group. Four weeks after the last immunization, the animals were challenged with 10(4) P. falciparum-parasitized erythrocytes. Peak levels of parasitemia were lower in the 16 monkeys that received region II-containing plasmids or proteins than in the 16 controls (geometric mean: 194,178 and 410,110 parasites/microL, respectively; P=.013, Students t test). Three of 4 monkeys in the Dv-Pv group did not require treatment. These data demonstrate that immunization with EBA-175 region II induces a significant antiparasite effect in vivo.

A dengue virus serotype-1 DNA vaccine induces virus neutralizing antibodies and provides protection from viral challenge in Aotus monkeys.

A DNA vaccine that expresses the premembrane/membrane (prM) and envelope (E) genes of dengue virus serotype-1 was tested for immunogenicity and protection against dengue-1 virus challenge in Aotus nancymae monkeys. The vaccine, in 1 mg doses, was administered intradermally (i.d.) to three monkeys and intramuscularly (i.m.) to three others. For controls, a 1 mg dose of vector DNA was administered i.d. to two monkeys and i.m. to one. All animals were primed and then boosted at one and five months post priming. Sera were collected monthly and analyzed for dengue-1 antibodies by enzyme linked immunosorbent assay (ELISA) and plaque reduction neutralization test (PRNT). Dengue-1 antibodies were detectable in the sera from i.d. and i.m. vaccine inoculated animals one month after the first boost and peaked one month after the second boost. The antibody levels from sera of animals that received the vaccine via the i.d. route were twice those from sera of animals that received the vaccine via the i.m. route. Six months after the second boost all inoculated and two naive monkeys were challenged with 1.25x10(4) plaque forming units (PFU) of dengue-1 virus. Two vaccine immunized animals were protected from viremia while the others showed a reduction in viremia. The mean days of viremia were 1 and 1.3 for the animals that were immunized with the vaccine via the i.d. or i.m. route, respectively vs 4 and 2 mean days of viremia in the animals inoculated with control DNA. Naive animals were viremic for an average of 4 days. All of the three control monkeys that received control DNA inoculum by either the i.d. or i.m. route had an intermittent viremia pattern with one or more negative days interspersed between the positive days. This pattern was not observed in any of the vaccine recipients or the naïve control monkeys. These results demonstrate that DNA immunization is a promising approach for the development of dengue vaccines and that A. nancymae monkeys are suitable for dengue vaccine trials.

Cross-serotype neutralization of dengue virus in Aotus nancymae monkeys.

Previously, we observed that serum from humans immune to dengue serotype 1 (dengue-1) neutralized the American genotype of dengue serotype 2 (American-2) to a greater extent than it neutralized the Asian genotype of dengue serotype 2 (Asian-2). To determine if this activity is protective, Aotus nancymae monkeys were infected with dengue-1 followed by either American-2 or Asian-2. Dengue-1-infected animals produced antibody with neutralizing titers of 2656 antibodies against dengue-1, 409 against American-2, and <20 against Asian-2. Infection with American-2 did not produce detectable viremia in either dengue-1-immune or dengue-1-naive animals. These findings support the hypothesis that dengue-1 immunity might have prevented disease or altered the severity of disease in individuals sequentially infected with dengue-1 and American-2.

Prevalence of antibody to Plasmodium falciparum antigens among feral Saimiri monkeys in the Amazon basin region of Peru

The squirrel monkey (Saimiri spp.) is a small New World monkey that has been used for several years in biomedical investigations. Recently, this monkey was reported as a model for evaluating candidate vaccines for protection against Plasmodium falciparum infection [5,7,8,11]. However, it is commonly assumed that these monkeys are resistant to P. falciparum infection and that splenectomy is required to render them susceptible to infection [5,14]. However, as described in this report, because of serological evidence of natural falciparum infection in blood samples obtained from feral Saimiri sciureus macrodon, it is suggested to prescreen these monkeys for antibodies before using them in malaria studies.