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      Safety, tolerability, and immunogenicity of two Zika virus DNA vaccine candidates in healthy adults: randomised, open-label, phase 1 clinical trials

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      , MD a , , , PhD a , , , PhD a , , PhD b , , MSc a , , PhD a , , MPH a , , RN a , , BSN a , , RN a , , CRNP a , , FNP a , , RN a , , PhD a , , MD d , , MD e , , MD d , , MD e , , CRNP e , , MSc a , , PhD a , , MS c , , MS c , , PhD c , , MS c , , MS c , , PhD c , , PhD a , , PhD a , , Sen NIH Investigator, MD a , , MD a , , Sen NIH Investigator, PhD c , , DO a , , MD a , * , VRC 319, VRC 320 study teams
      Lancet (London, England)
      Elsevier Ltd.

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          Summary

          Background

          The Zika virus epidemic and associated congenital infections have prompted rapid vaccine development. We assessed two new DNA vaccines expressing premembrane and envelope Zika virus structural proteins.

          Methods

          We did two phase 1, randomised, open-label trials involving healthy adult volunteers. The VRC 319 trial, done in three centres, assessed plasmid VRC5288 (Zika virus and Japanese encephalitis virus chimera), and the VRC 320, done in one centre, assessed plasmid VRC5283 (wild-type Zika virus). Eligible participants were aged 18–35 years in VRC19 and 18–50 years in VRC 320. Participants were randomly assigned 1:1 by a computer-generated randomisation schedule prepared by the study statistician. All participants received intramuscular injection of 4 mg vaccine. In VRC 319 participants were assigned to receive vaccinations via needle and syringe at 0 and 8 weeks, 0 and 12 weeks, 0, 4, and 8 weeks, or 0, 4, and 20 weeks. In VRC 320 participants were assigned to receive vaccinations at 0, 4, and 8 weeks via single-dose needle and syringe injection in one deltoid or split-dose needle and syringe or needle-free injection with the Stratis device (Pharmajet, Golden, CO, USA) in each deltoid. Both trials followed up volunteers for 24 months for the primary endpoint of safety, assessed as local and systemic reactogenicity in the 7 days after each vaccination and all adverse events in the 28 days after each vaccination. The secondary endpoint in both trials was immunogenicity 4 weeks after last vaccination. These trials are registered with ClinicalTrials.gov, numbers NCT02840487 and NCT02996461.

          Findings

          VRC 319 enrolled 80 participants (20 in each group), and VRC 320 enrolled 45 participants (15 in each group). One participant in VRC 319 and two in VRC 320 withdrew after one dose of vaccine, but were included in the safety analyses. Both vaccines were safe and well tolerated. All local and systemic symptoms were mild to moderate. In both studies, pain and tenderness at the injection site was the most frequent local symptoms (37 [46%] of 80 participants in VRC 319 and 36 [80%] of 45 in VRC 320) and malaise and headache were the most frequent systemic symptoms (22 [27%] and 18 [22%], respectively, in VRC 319 and 17 [38%] and 15 [33%], respectively, in VRC 320). For VRC5283, 14 of 14 (100%) participants who received split-dose vaccinations by needle-free injection had detectable positive antibody responses, and the geometric mean titre of 304 was the highest across all groups in both trials.

          Interpretation

          VRC5283 was well tolerated and has advanced to phase 2 efficacy testing.

          Funding

          Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health.

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          Most cited references21

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          Zika Virus Infection as a Cause of Congenital Brain Abnormalities and Guillain–Barré Syndrome: Systematic Review

          Background The World Health Organization (WHO) stated in March 2016 that there was scientific consensus that the mosquito-borne Zika virus was a cause of the neurological disorder Guillain–Barré syndrome (GBS) and of microcephaly and other congenital brain abnormalities based on rapid evidence assessments. Decisions about causality require systematic assessment to guide public health actions. The objectives of this study were to update and reassess the evidence for causality through a rapid and systematic review about links between Zika virus infection and (a) congenital brain abnormalities, including microcephaly, in the foetuses and offspring of pregnant women and (b) GBS in any population, and to describe the process and outcomes of an expert assessment of the evidence about causality. Methods and Findings The study had three linked components. First, in February 2016, we developed a causality framework that defined questions about the relationship between Zika virus infection and each of the two clinical outcomes in ten dimensions: temporality, biological plausibility, strength of association, alternative explanations, cessation, dose–response relationship, animal experiments, analogy, specificity, and consistency. Second, we did a systematic review (protocol number CRD42016036693). We searched multiple online sources up to May 30, 2016 to find studies that directly addressed either outcome and any causality dimension, used methods to expedite study selection, data extraction, and quality assessment, and summarised evidence descriptively. Third, WHO convened a multidisciplinary panel of experts who assessed the review findings and reached consensus statements to update the WHO position on causality. We found 1,091 unique items up to May 30, 2016. For congenital brain abnormalities, including microcephaly, we included 72 items; for eight of ten causality dimensions (all except dose–response relationship and specificity), we found that more than half the relevant studies supported a causal association with Zika virus infection. For GBS, we included 36 items, of which more than half the relevant studies supported a causal association in seven of ten dimensions (all except dose–response relationship, specificity, and animal experimental evidence). Articles identified nonsystematically from May 30 to July 29, 2016 strengthened the review findings. The expert panel concluded that (a) the most likely explanation of available evidence from outbreaks of Zika virus infection and clusters of microcephaly is that Zika virus infection during pregnancy is a cause of congenital brain abnormalities including microcephaly, and (b) the most likely explanation of available evidence from outbreaks of Zika virus infection and GBS is that Zika virus infection is a trigger of GBS. The expert panel recognised that Zika virus alone may not be sufficient to cause either congenital brain abnormalities or GBS but agreed that the evidence was sufficient to recommend increased public health measures. Weaknesses are the limited assessment of the role of dengue virus and other possible cofactors, the small number of comparative epidemiological studies, and the difficulty in keeping the review up to date with the pace of publication of new research. Conclusions Rapid and systematic reviews with frequent updating and open dissemination are now needed both for appraisal of the evidence about Zika virus infection and for the next public health threats that will emerge. This systematic review found sufficient evidence to say that Zika virus is a cause of congenital abnormalities and is a trigger of GBS.
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            Rapid development of a DNA vaccine for Zika virus.

            Zika virus (ZIKV) was identified as a cause of congenital disease during the explosive outbreak in the Americas and Caribbean that began in 2015. Because of the ongoing fetal risk from endemic disease and travel-related exposures, a vaccine to prevent viremia in women of childbearing age and their partners is imperative. We found that vaccination with DNA expressing the premembrane and envelope proteins of ZIKV was immunogenic in mice and nonhuman primates, and protection against viremia after ZIKV challenge correlated with serum neutralizing activity. These data not only indicate that DNA vaccination could be a successful approach to protect against ZIKV infection, but also suggest a protective threshold of vaccine-induced neutralizing activity that prevents viremia after acute infection.
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              Zika virus and microcephaly: why is this situation a PHEIC?

              When the Director-General of WHO declared, on Feb 1, 2016, that recently reported clusters of microcephaly and other neurological disorders are a Public Health Emergency of International Concern (PHEIC), 1 it was on the advice of an Emergency Committee of the International Health Regulations and of other experts whom she had previously consulted. We are the members of the Emergency Committee, and we were identified by the Director-General from rosters of experts that had been submitted by WHO Member States. Our advice to declare a PHEIC was not made on the basis of what is currently known about Zika virus infection. During our discussions it became clear that infection with the Zika virus, unlike other arbovirus infections including dengue and chikungunya, causes a fairly mild disease with fever, malaise, and at times a maculopapular rash, conjunctivitis, or both. 2 Additional information from previous outbreaks suggested that about 20% of people infected with Zika virus develop these symptoms, and that the rest are asymptomatic. 2 Fatality from Zika virus infection is thought to be rare. 2 Our advice to declare a PHEIC was rather made on the basis of what is not known about the clusters of microcephaly, Guillain-Barré syndrome, and possibly other neurological defects reported by country representatives from Brazil and retrospectively from French Polynesia that are associated in time and place with outbreaks of Zika infection.3, 4 The Emergency Committee meeting was convened rapidly by WHO. We were contacted by the Director-General 4 days before the Emergency Committee meeting, and by the time we met WHO had thoroughly prepared the meeting. At the start of the meeting, the WHO legal counsel provided three criteria to help the Emergency Committee decide whether the present situation was a PHEIC. A PHEIC must: (1) constitute a health risk to other countries through international spread; (2) potentially require a coordinated response because it is unexpected, serious, or unusual; and (3) have implications beyond the affected country that could require immediate action. Representatives from four countries (Brazil, El Salvador, France, and the USA) that have had either outbreaks or importations of Zika virus, and a group of arbovirus specialists, took part in the meeting. Some of them had been working for the past months with the WHO Regional Office in the Americas on the Zika virus outbreaks, and before that on those caused by the dengue and chikungunya viruses. During one country representative's account of Zika virus in French Polynesia, robust and convincing retrospective data were presented about an increase in neurological disorders during the period when there was an outbreak of Zika virus. Other presentations described current clusters of microcephaly and limited information about Zika virus identified in fetuses or infants, pointing out the temporal association with circulation of the Zika virus. After these country presentations, and comments by the assembled arbovirologists, we were able to discern as a committee, and then agree unanimously in an initial poll, that the clusters of microcephaly and neurological disorders, and their possible association with the Zika virus, constituted a PHEIC. Upon further discussion, it became clear that there was no standard surveillance case definition for microcephaly. The first recommendation of the PHEIC was to call for standardised and enhanced surveillance of microcephaly in areas of known Zika virus transmission. Such surveillance is not only important in countries where there are current and recent outbreaks, but is also retrospectively relevant in African and Asian countries where outbreaks have been occurring since the Zika virus was first identified in 1947.5, 6 Further, we felt that surveillance data should become available within months. © 2016 Fabrice Coffrini 2016 Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. Our second recommendation under the PHEIC is for increased research into the aetiology of confirmed clusters of microcephaly and neurological disorders to determine whether there is a causative link to Zika virus, other factors, and cofactors. Neurological fetal defects occur with other viral infections such as rubella, which are preventable by vaccine, 7 and could also be caused by factors such as exposure to chemicals or toxins and other environmental factors.8, 9 We understood that this PHEIC recommendation will take much longer to implement than surveillance, and will require accumulation of scientific evidence from post-mortem analyses, case-control studies, and other studies as recommended by experts in microcephaly, obstetric and neonatal medicine, and public health. Part of our discussion also included the need for development of an animal model, and of the possibility of eventually proving Koch's postulates. After our discussion on the PHEIC, there was unanimous agreement to make recommendations for precautionary measures to prevent arboviral infection. In addition to being good public health practice, which would be intensified should the clusters of microcephaly and other neurological disorders be linked to the Zika virus, they should also result in the prevention of chikungunya and dengue outbreaks.10, 11, 12 Among those recommendations were the need for: stronger surveillance of Zika virus infection with the rapid development and sharing of diagnostics suitable for seroprevalence studies and that do not require antigen presence; improved communication about the risks of outbreaks of Zika and other arboviruses; implementation of vector control measures to decrease exposure to bites from the Aedes aegypti mosquito; and guidance to be available to pregnant women so that they better understand the present situation and are empowered to make a decision about personal protection and pregnancy. We also provided longer-term advice to the Director-General to continue discussions with vaccine developers and regulatory agencies that WHO had already begun, to provide regular and clear guidance on risks associated with travel, and to ensure that all countries share data as they work with WHO to address the recommendations of the PHEIC. Since the Emergency Committee meeting we have continued to communicate among ourselves, and our hope is that WHO will work in the way that successfully led to control of the outbreak of severe acute respiratory syndrome (SARS) in 2003 when WHO established virtual networks of experts around the world who worked by telephone and the internet to collaborate in surveillance, clinical management, and research.13, 14, 15 The networks established during the SARS outbreak worked in environments that provided the confidentiality and security necessary to freely share data used for improving public health. With policies recently developed by The Lancet and other medical journals to accept for publication data that may have previously been shared openly for better outbreak prevention and control, we believe that there should be no excuse for not creating such an environment for sharing of data collected under the PHEIC.16, 17 Since the Director-General declared the PHEIC on microcephaly and neurological disorders, many of us have had questions about how our recommendation relates to the PHEIC called by the Director-General for the 2014 Ebola outbreaks in west Africa based on the recommendation of a different Emergency Committee. The answer to us is clear. The Director-General declared the Ebola outbreaks a PHEIC because of what science knew about the Ebola virus from many years of research during outbreaks in the past, whereas she declared the current PHEIC because of what is not known about the current increase in reported clusters of microcephaly and other disorders, and how this might relate to concurrent Zika outbreaks. We were told by the Director-General that she would convene us again within 3 months to reassess the situation, as required under the International Health Regulations. We are confident that virtual meetings will allow us to review global collective action and to learn from WHO about progress in understanding the present situation of microcephaly and neurological disorders and progress in implementation of the precautionary and preparatory measures related to Zika.
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                Author and article information

                Contributors
                Journal
                Lancet
                Lancet
                Lancet (London, England)
                Elsevier Ltd.
                0140-6736
                1474-547X
                5 December 2017
                10-16 February 2018
                5 December 2017
                : 391
                : 10120
                : 552-562
                Affiliations
                [a ]Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
                [b ]Biostatistics Research Branch, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
                [c ]Viral Pathogenesis Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
                [d ]Department of Medicine, Division of Infectious Diseases, Hope Clinic of the Emory Vaccine Center, Emory School of Medicine, Decatur, GA, USA
                [e ]University of Maryland Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, MD, USA
                Author notes
                [* ]Correspondence to: Dr Grace L Chen, Vaccine Research Center, Clinical Trials Program, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA grace.chen@ 123456nih.gov
                [†]

                Contributed equally

                Article
                S0140-6736(17)33105-7
                10.1016/S0140-6736(17)33105-7
                6379903
                29217376
                ae6226f6-504c-4b0b-b565-f32c00593f2b
                © 2017 Elsevier Ltd. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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