key: cord-0275309-92a79gnl authors: van Doremalen, Neeltje; Avanzato, Victoria A.; Feldmann, Friederike; Schulz, Jonathan E.; Haddock, Elaine; Okumura, Atsushi; Lovaglio, Jamie; Hanley, Patrick W.; Cordova, Kathleen; Saturday, Greg; Lambe, Teresa; Gilbert, Sarah C.; Munster, Vincent J. title: ChAdOx1 NiV vaccination protects against lethal Nipah Bangladesh virus infection in African green monkeys date: 2021-07-20 journal: bioRxiv DOI: 10.1101/2021.07.20.452991 sha: 7389a1794b37ddfcf7630f3c140cf94c274a6485 doc_id: 275309 cord_uid: 92a79gnl Nipah virus (NiV) is a highly pathogenic and re-emerging virus which causes sporadic but severe infections in humans. Currently, no vaccines against NiV have been approved. We previously showed that ChAdOx1 NiV provides full protection against a lethal challenge with NiV Bangladesh (NiV-B) in hamsters. Here, we investigated the efficacy of ChAdOx1 NiV in the lethal African green monkeys (AGMs) NiV challenge model. AGMs were vaccinated either 4 weeks before challenge (prime vaccination), or 8 and 4 weeks before challenge with ChAdOx1 NiV (prime-boost vaccination). A robust humoral and cellular response was detected starting 14 days post initial vaccination. Upon challenge, control animals displayed a variety of signs and had to be euthanized between 5- and 7-days post inoculation. In contrast, vaccinated animals showed no signs of disease, and we were unable to detect infectious virus in all but one swab and all tissues. Importantly, no to limited antibodies against fusion protein or nucleoprotein IgG could be detected 42 days post challenge, suggesting that vaccination induced a very robust protective immune response preventing extensive virus replication. One Sentence Summary A single vaccination with ChAdOx1 NiV protects African green monkeys against lethal disease induced by Nipah virus inoculation. Nipah virus (NiV) is a highly pathogenic re-emerging member of the Paramyxovirus family, 16 genus Henipavirus. NiV causes sporadic infections in humans, resulting in severe neurological 17 and respiratory disease, often with a fatal outcome. NiV was first detected in 1998, when the 18 strain NiV-Malaysia (NiV-M) caused an outbreak of severe encephalitis in pig farmers from 19 Malaysia and Singapore, with a case-fatality rate of 38% 1 . Since 2001, outbreaks with a closely 20 related strain, NiV-Bangladesh (NiV-B), have occurred almost yearly in Bangladesh 2 , resulting 21 in 319 accumulated cases and 225 associated deaths (case-fatality rate 71%) 3 . Most recently, 22 outbreaks have also been reported in India 4 . 23 The natural reservoir of NiV is the Pteropus spp. fruit bat [5] [6] [7] . Outbreaks in Bangladesh and India 24 have been associated with the consumption of date palm sap, which may have been contaminated 25 with bat urine [8] [9] [10] . In contrast, in Malaysia and Singapore, pigs were the intermediate host, likely 26 infected via the consumption of mango fruits, which were contaminated with NiV after partial 27 consumption by bats 11 . Importantly, human-to-human transmission of NiV has also been 28 reported 12, 13 . 29 Although the total number of cases caused by NiV are limited, the virus causes severe disease 30 and transmits between humans 12, 13 , and can infect a wide range of animals 14 , and thus NiV is 31 categorized by the WHO as a pathogen with epidemic potential which poses a great public health 32 risk and requires research aimed at the development of countermeasures. 33 Several vaccine candidates have been evaluated in animal models 15 . The most extensively 34 studied vaccine to date is based on the glycoprotein of Hendra virus (HeV), another member of 35 the genus Henipavirus 16 . HeV-sG, a soluble form of the HeV receptor binding glycoprotein, was 36 marketed by Zoetis, Inc., in Australia as an equine vaccine against HeV under the name 37 Equivac ® HeV. It is the first commercialized vaccine against a BSL-4 agent 17 . 38 Recently, it was shown that HeV-sG vaccination can protect African green monkeys (AGMs) 39 against lethal NiV disease as early as 7 days post immunization 17 . Enrollment has started for a 40 Phase I randomized placebo-controlled clinical trial on NiV-vaccine candidate HeV-sG-V, which 41 is based on HeV-sG, with results expected in October 2021 (ClinicalTrials.gov NCT04199169). 42 This clinical trial is the first of its kind for NiV and is the result of a global partnership between 43 Auro Vaccines LLC and the Coalition of Epidemic Preparedness Innovations (CEPI). 44 In the current study, we are testing efficacy of a different NiV vaccine candidate in AGMs. 45 ChAdOx1 is a replication-deficient simian adenoviral vector, which has been developed for a 46 multitude of different pathogens by the University of Oxford. A vaccine based on this vector 47 named ChAdOx1 nCoV-19 (also known as AZD1222, Vaxzevria, or Covishield) has been 48 developed against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the 49 etiological agent of COVID-19. ChAdOx1 nCoV-19 has been fully approved in Brazil and 50 approved for emergency use in 64 additional countries. The effectiveness of the vaccine is 92% 51 and is 79% against symptomatic infection 18, 19 . 52 We have previously demonstrated that a single dose of ChAdOx1 NiV, which encodes the 53 receptor binding protein (G) of NiV-B, fully protected Syrian hamsters against a lethal dose of 54 NiV-B or NiV-M 20 . In the current study, we investigate whether ChAdOx1 NiV is protective in 55 the lethal NiV AGM model 21 . We show that a single dose of ChAdOx1 NiV results in a robust 56 innate and adaptive immune response, which is fully protective against lethal disease in AGMs. 57 Furthermore, no to limited immune response against nucleoprotein or fusion protein was 58 detected in vaccinated animals post challenge, but was detected in control challenged animals, 59 suggesting that vaccination provides near complete protection against NiV infection. (Table S1 ). All four animals in this group reached an 94 endpoint clinical score of 35 or higher between 5 to 7 DPI and were euthanized. In contrast, no 95 signs of disease were observed in animals vaccinated with ChAdOx1 NiV (Figure 2A -B, Table 96 S1). Exams were performed on 0, 3, 5, 7, 10, 14, 21, 28, 35, and 42 DPI. Radiographs were 97 scored as previously described 22 . Whereas no to limited changes from baseline were observed in 98 ChAdOx1 NiV vaccinated animals (score between 0-2), radiograph scores of control-vaccinated 99 animals started increasing at 3 DPI (score between 0-3) and continued to increase until day of 100 necropsy (score between 7-14, Figure 2C ). Throat and nose swabs were collected on all exam Figure 4C ). In situ hybridization reveals abundant viral RNA distributed throughout lesions in 143 vascular endothelium and type I&II pneumocytes in tissue from control animals ( Figure 4F ). 144 Infectious virus was only detected in tissue obtained from control animals, and not in tissue 145 obtained from vaccinated animals ( Figure 3D , S1). TCID50/mL). Animals were scored daily by the same person who was blinded to study group allocations 234 using a standardized scoring sheet 37 . Scoring was based on the following criteria: general appearance, 235 skin and coat appearance, discharge, respiration, feces and urine appearance, appetite, and activity. After blocking with 5% milk in PBS with 0.01% tween (PBST), serum in 5% milk in PBST was 311 incubated at RT for 1 hr. Antibodies were detected using affinity-purified antibody peroxidase-labeled 312 goat-anti-monkey IgG (Seracare) in 5% milk in PBST and TMB 2-component peroxidase substrate 313 (Seracare) and read at 450 nm. 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