We will examine and compare acute infection (14 days) with the ZIKV strain PRVABC59, a wild type (WT) clone based on this strain, and a barcoded clone. Wild type ZIKV strain PRVABC59 was originally isolated by CDC from human serum and will be used to infect three nonpregnant rhesus macaques. These three animals will serve as controls for comparison with six additional animals, three which will be infected with a wild type (WT) ZIKV clone and three which will be infected with a barcoded ZIKV clone, both generated by Greg Ebel’s group at Colorado State University (CSU). These clones were produced using a bacteria-free system and were rescued using a CMV promoter in Vero cells; both the WT clone and the barcoded clone for these infections are passage 0, direct from transfection. These experiments are the first infections with these clones in NHP and will help characterize the in vivo growth characteristics and pathology of the ZIKV clones in an immune competent NHP host.
In addition, the 3 animals infected with the wildtype ZIKV strain will be subjected to mosquito feeding by Aedes aegypti (Black-eyed Liverpool strain) at 3 dpi to test transmissibility of this Zika virus strain from Indian rhesus macaques to mosquitoes.
Nine Indian-origin rhesus macaques, three animals in each group, will be infected subcutaneously with 10E4 PFU ZIKV Puerto Rico (PRVABC59), WT ZIKV clone, or barcoded clone. Blood plasma, urine, and saliva will be collected from each animal at each timepoint (0-14 dpi), and serum will be collected at 0 and 14 dpi. Viral RNA will be measured using qRT-PCR for all sample types. The viral load trajectories and viremias of these animals will be compared between groups.
- 634675 was challenged with 10E4 PFU WT ZIKV-PRVABC59 virus
- 566628 was challenged with 10E4 PFU WT ZIKV-PRVABC59 virus
- 311413 was challenged with 10E4 PFU WT ZIKV-PRVABC59 virus
- 296198 was challenged with 10E4 PFU WT ZIKV clone (PRVABC59 background)
- 118693 was challenged with 10E4 PFU WT ZIKV clone (PRVABC59 background)
- 962498 was challenged with 10E4 PFU WT ZIKV clone (PRVABC59 background)
- 514982 was challenged with 10E4 PFU barcoded ZIKV clone (PRVABC59 background)
- 715132 was challenged with 10E4 PFU barcoded ZIKV clone (PRVABC59 background)
- 688387 was challenged with 10E4 PFU barcoded ZIKV clone (PRVABC59 background)
January 9, 2017: Inoculation of all nine animals was successful.
January 10, 2017: Animals that received the barcoded clone inoculation have some redness and irritation at the site of inoculation ranging from mild redness to redness. It is not clear whether this irritation/redness is due to shaving of the site prior to infection or is related to the infection.
January 19, 2017: To examine vertebrate host-mosquito transmission from animals infected with the WT PRVABC59 strain, Aedes aegypti (Black-eyed Liverpool strain) were fed on three animals (634675, 566628, and 311413) on January 12, 2017 (3 dpi).
Clinical and Assay Data
Viral RNA quantification
- Plasma viral load Chart
- Pan urine viral load Chart
- Oral swab viral load Chart