Zika Summit abstracts - May 2016

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ZIKA virus replication in human placenta

T. Couderc2-1, G. Gessain2-1, M. Lavina2-1, C. Charlier-Woerther2-3, O. Disson2-1, M. Lecuit2-1-3 1U1117, Inserm 2Biology of Infections, Institut Pasteur 3Paris Descartes University, Paris, France

Zika virus (ZIKV) is an emerging arbovirus of the Flaviviridae family transmitted to humans by mosquito bites and sexual route. ZIKV caused a major outbreak in French Polynesia in 2013/2014 and has since spread in 2015 to the Americas, notably in Brazil. Most cases of zika infection in otherwise healthy adults are asymptomatic or pauci-symptomatic, with fever, rash, arthralgia, and conjunctivitis. ZIKV infection has recently been associated with severe fetopathy, including microcephaly (Brasil et al., N Engl J Med. 2016, in press). Placental calcifications are observed in ZIKV-infected pregnant women and the virus is also present in the amniotic fluid and brain of fetuses with microcephaly (Calvet et al., Lancet Infect. Dis., 2016, in press; Mlakar et al., N Engl J Med. 2016, in press; Sarno et al., PLOS Negl Trop Dis, 2016).

Together, these data strongly suggest that ZIKV has ability to cross the placental barrier and induce fetopathy.
Here, we have assessed the ability of ZIKV to infect human placental explants and human trophoblast cell lines, as compared to Chikungunya virus (CHIKV), another emerging arbovirus which is not able to infect the placenta and is exclusively transmitted vertically from viremic mother to their baby during parturition via placental breaches. We infected third trimester human placental explants with either ZIKV or CHIKV, and viral load in placental tissues and supernatants were determined daily from day 1 to day 4 post-infection. We found that ZIKV replicates in human placental explants, as shown by viral titers in placental tissues and supernatants, while, as expected, CHIKV does not. The identification of the cells targeted by ZIKV in human placental explants is currently investigated. In addition, a human trophoblast cell line was shown to be susceptible to ZIKV infection, but resistant to CHIKV infection. These preliminary data suggest a direct ZIKV tropism for human placenta. The relevance of these results will be assessed by detection of ZIKV in human placental biopsies of infected pregnant women. Studies in animal models susceptible to ZIKV will help to decipher how and when this virus crosses the placental barrier.
Deciphering the placental phase of ZIKV infection is required to understand its vertical transmission and develop potential preventive and therapeutic strategies against this emerging teratogenic arbovirus.

Evaluation of immune response in human Zika virus infection

L. Gois2, MFR. Grassi2, B. Galvao Castro2, G. Oliveira2, SM. Arruda2, G.S. Campos4, S. Sardi4, A. Bandeira3, B. Autran1, V. Vieillard1, C. Combadiere1, H. Yssel1, G. Gorochov1
1Universite Pierre etMarie Curie Paris / Centre de Recherches en Immunologie etMaladies infectieuses, CIMI-Paris, Franga 2Centro de Pesquisa Gongalo Moniz (CPqGM) /Laboratorio Avangado de Saude Publica (LASP), Fundagao Oswaldo Cruz (FIOCRUZ) 3Secretaria de Saude da Bahia 4Laboratorio de Virologia do Instituto de Ciencias da Saude, Universidade Federal da Bahia, Salvador, Brazil

Zika virus (ZIKV) infection in Brazil, was first described in 2015 and it overlapped with a significant increase in number of cases of Guillain-Barre syndrome in adults and also in neurological malformation in infants, including microcephaly. The role of the immune response in protection and/or development of severe cases of infectionremain to be addressed. This proposal aims to evaluate both the innate and acquired immune response in patients with different clinical forms of the infection. Especially seeks to identify specific immune signatures to different clinical forms of the disease and, in addition, assess whether there are cross-reactivities to virus chikungunya and dengue (DENV). To this, a consortium gatheringclinical investigators (Department of Bahia State Health), virologists (Federal University of Bahia), immunologists (Fiocruz, Bahia) and molecular biologists and immunologists (Departement d'lmmunologie Hopitaux Universitaires Pitie- Salpetriere, Universite Pierre et Marie Curie - France) is being formed. The lymphocyte responses to ZIKV peptides and recombinant proteins as well as the innate immune cell responsesto the virus will be evaluated using different methodologies, including mass cytometry (CyTOF) that simultaneously analyzes up to 30 parameters in a single tube. Antibodies from ZIKV-infected individuals will be evaluated in relation to antigenic specificity, neutralizing capacity andcross-reactivity with DENV-4 and chikungunya virus antigens. The results of this project would contribute to the understanding of pathogenesis of the disease, in addition to the characterization of dominant immune response that will be important for the development of diagnostic tests, vaccines and identification of potential drug targets.

ZIKA virus replication in human placenta

M. Lecuit, Biology of Infection Unit, Institut Pasteur, Paris, France

Zika virus (ZIKV) is an emerging arbovirus of the Flaviviridae family transmitted to humans by mosquito bites and sexual route. ZIKV caused a major outbreak in French Polynesia in 2013/2014 and has since spread in 2015 to the Americas, notably in Brazil. Most cases of zika infection in otherwise healthy adults are asymptomatic or pauci-symptomatic, with fever, rash, arthralgia, and conjunctivitis. ZIKV infection has recently been associated with severe fetopathy, including microcephaly (Brasil et al., N Engl J Med. 2016, in press). Placental calcifications are observed in ZIKV-infected pregnant women and ZIKV is also present in the amniotic fluid and brain of fetuses with microcephaly (Calvet et al., Lancet Infect. Dis., 2016, in press; Mlakar et al., N Engl J Med. 2016, in press; Sarno et al., PLOS Negl Trop Dis, 2016). Together, these data strongly suggest that ZIKV has ability of ZIKV to cross the placental barrier and induce fetopathy.

Here, we have assessed the ability of ZIKV to infect human placental explants and human trophoblast cell lines, as compared to Chikungunya virus (CHIKV), another emerging arbovirus which is not able to infect the placenta and is exclusively transmitted vertically from viremic mother to their baby during parturition via placental breaches. We infected third trimester human placental explants with either ZIKV or CHIKV, and viral load in placental tissues and supernatants were determined daily from day 1 to day 4 post-infection. We found that ZIKV replicates in human placental explants, as shown by viral titers in placental tissues and supernatants, while, as expected, CHIKV does not. The identification of the cells targeted by ZIKV in human placental explants is currently investigated. In addition, a human trophoblast cell line was shown to be susceptible to ZIKV infection, but resistant to CHIKV infection. These preliminary data suggest a direct ZIKV tropism for human placenta. The relevance of these results will be assessed by detection of ZIKV in human placental biopsies of infected pregnant women. Studies in animal models susceptible to ZIKV will help decipher how and when this virus crosses the placental barrier.
Understanding the placental phase of ZIKV infection is required to understand its vertical transmission and develop potential preventive and therapeutic strategies against this emerging teratogenic arbovirus.

Zika Virus Infection in Pregnant Women in Rio de Janeiro Preliminary Report

P. Brasil. JP. Pereira Junior, C. Raja Gabaaglia , L. Damasceno, M. Wakimoto , RM. Ribeiro Nogueira , P. Carvalho De Sequeira1, A. Machado Siqueira1, LM. Abreu De Carvalho1, D. Cotrim Da Cunha1, GA. Calvet1, ES. Neves1, ME.
Moreira , AE. Rodrigues Baiao , PR. Nassar De Carvalho, C. Janzen , SG. Valderramos , JD. Cherry , AM. Bispo De Filippis1, K. Nielsen-Saines2
1Fundacao Oswaldo Cruz, Rio De Janeiro, Brazil 2David Geffen UCLA School of Medicine, Los Angeles (C.J., S.G.V., J.D.C., K.N.-S.), Los Angeles Biomedical Research Institute of Southern California, Oceanside (C.R.G.), San Diego, United States

—Background Zika virus (ZIKV) has been linked to neonatal microcephaly. To characterize the spectrum of ZIKV disease in pregnancy, we followed patients in Rio de Janeiro to describe clinical manifestations in mothers and repercussions of acute ZIKV infection in fetuses. Methods We enrolled pregnant women in whom a rash had developed within the previous 5 days and tested blood and urine specimens for ZIKV by reverse-transcriptase-polymerase-chain-reaction assays. We followed the women prospectively and collected clinical and ultrasonographic data. Results A total of 88 women were enrolled from September 2015 through February 2016; of these 88 women, 72 (82%) tested positive for ZIKV in blood, urine, or both. The timing of acute ZIKV infection ranged from 5 to 38 weeks of gestation. Predominant clinical features included pruritic descending macular or maculopapular rash, arthralgias, conjunctival injection, and headache; 28% had fever (short-term and low-grade). Women who were positive for ZIKV were more likely than those who were negative for the virus to have maculopapular rash (44% vs. 12%, P=0.02), conjunctival involvement (58% vs. 13%, P=0.002), and lymphadenopathy (40% vs. 7%, P=0.02). Fetal ultrasonography was performed in 42 ZIKV-positive women (58%) and in all ZIKV-negative women. Fetal abnormalities were detected by Doppler ultrasonography in 12 of the 42 ZIKV-positive women (29%) and in none of the 16 ZIKV-negative women. Adverse findings included fetal deaths at 36 and 38 weeks of gestation (2 fetuses), in utero growth restriction with or without microcephaly (5 fetuses), ventricular calcifications or other central nervous system (CNS) lesions (7 fetuses), and abnormal amniotic fluid volume or cerebral or umbilical artery flow (7 fetuses). To date, 8 of the 42 women in whom fetal ultrasonography was performed have delivered their babies, and the ultrasonographic findings have been confirmed. Conclusions Despite mild clinical symptoms, ZIKV infection during pregnancy appears to be associated with grave outcomes, including fetal death, placental insufficiency, fetal growth restriction, and CNS injury. (NEJM March 4, 2016 DOI: 10.1056/NEJMoa1602412). From the analyzed period to April 2016 data will be updated.

Transient Hearing Loss in Adults Associated with Zika Virus Infection

ES. Vinhaes4, L. Santos3. L. Dias4, NA. Andrade4, VH. Bezerra4, L. De Moraes3, SG. Rodrigues2, N. Vasilakis6, AI. Ko7- 3, BB. Andrade3, IC. Siqueira3, R. Khouri3-1, VS. Boaventura3-5 7 0
laboratory of Clinical and Epidemiological Virology, Rega Institute for Medical Research, Leuven, Belgium 2Instituto Evandro Chagas, Belem 3Centro de Pesquisa Gongalo Muniz, Laboratorio de Imunoparasitologia, Fundagao Oswaldo Cruz (Fiocruz), Lauro De Freitas 4Hospital Santa Izabel 5Universidade Federal da Bahia, Salvador, Brazil 6Department of Pathology, Center for Biodefense and Emerging Infectious Diseases, University of Texas Medical Branch,
Galveston Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, United States
On May 2015 an outbreak of Zika virus (ZIKV) was identified in Brazil. Zika virus infection has been linked to neurological disorders like microcephaly and Guillain-Barre syndrome in Brazil and in other countries. Despite that, hearing impairment and pray-motor skills were not described. Here we report three cases admitted in the Emergency Unit with hearing loss, tinnitus and dizziness. All patients reported acute symptoms of itching exanthema, arthralgia and headache. Audiometry exams were performed before, during and after otologic symptoms in two patients and revealed temporary bilateral sensorineural hearing loss. A positive serologic test confirmed ZIKV infection in all patients. Our finding suggests a possibly association between Zika virus infection and acute hearing loss, which may persist for up to four weeks.

Epidemiologic Surveillance and Research Activities to Address Zika Virus and Pregnancy: An Update from the U.S. Centers for Disease Control and Prevention

C. Boyle2. M. Honein1, D. Jamieson1, C. Moore1, J. Staples1, S. Hillis1 2
Center on Birth Defects and Developmental Disabilities, U.S. Centers for Disease Control and Prevention, Atlanta, United States

The emergence of Zika virus in the Americas with its untoward impact on the developing fetus is resulting in a rapid response by the scientific and public health communities. The U.S. Centers for Disease Control and Prevention (CDC) responded by activating its Emergency Operations Center on January 22, 2016. Given the paucity of data specific to how the virus impacts the fetus, CDC has a launched a number of surveillance and research projects in the U.S. and with collaborators in other countries. On the domestic front, CDC has implemented the U.S. Zika Pregnancy Registry, an active surveillance network to prospectively enroll and follow pregnant women and their infants; a complementary registry has also been implemented in the Commonwealth of Puerto Rico. Both systems enroll pregnant women and infants with laboratory evidence of Zika virus infection, and prenatally and perinatally exposed infants. Individuals are enrolled into the registry by health departments or directly from health care providers. In Puerto Rico active surveillance is underway based on medical record abstraction. Data on the pregnant women will be collected at a number of points in pregnancy and on the infants at birth and at 2, 6, and 12 months of age, with follow-up to age 3 years in Puerto Rico. Clinical information will include attributes of the mother’s health and Zika virus testing during pregnancy, results of fetal and newborn evaluations, and medical and developmental information on the infant. Emerging data from the registries will be used to inform clinical recommendations and public health actions. CDC is also enhancing its ongoing state-based birth defects surveillance programs to identify, in a real-time manner, any infant born with microcephaly or other Zika-related birth defects. In the international arena, CDC is engaged in several research and surveillance activities in Central and South America. Most notably, we are collaborating with the Secretary of Health of Paraiba state, Brazil and the Brazilian Ministry of Health on a case-control study of microcephaly to further characterize the association with Zika virus infection and the clinical phenotype of fetal Zika infection. We are also collaborating with the Instituto Nacional de Salud in Colombia to enhance their national surveillance of Zika virus disease among pregnant women and to implement in-depth monitoring of pregnant women in selected regions of Colombia with high reported incidence of Zika virus disease. A description of CDC activities and collaborations will be presented, including current status and anticipated outcomes.

Detection of Zika virus rapidly, economically, locally and reliably without the need for a laboratory using Isothermal amplification

G. Aidelberg4. A. Kumar6, G. Gome7-4, M. Amelia Borba2, D. Bruneska Gondim Martins2, H. Castelletti1, J.
Albuquerque3
1'LIKA-Laboratory of Immunopathology Keizo Asami, Agronomic Institute of Pernambuco (IPA) 2LIKA-Laboratory of Immunopathology Keizo Asami, UFPE-Federal University of Pernambuco 3LIKA-Laboratory of Immunopathology Keizo Asami, UFRPE - Statistical and Informatics Department / Epitrack, Recife, Brazil 4GomeX labs 5WLW, Tel Aviv,
Israel 6University of Leicester; NIHR, Leicester, United Kingdom 7singularity university, Mountain View, United States
Zika virus (ZIKV) is an emerging arbovirus which can cause severe neurological disorders, spreading rapidly through the America, a public health emergency of international concern. Current diagnostic tests are inadequate, expensive and unavailable where they are needed most. We propose to develop, optimize and deploy an affordable, reliable, easy to use, rapid, and robust kit for rural and low-income use for the detection of arboviruses, both on human samples for diagnosis as well as on mosquitoes for surveillance purposes. The kit will utilize isothermal amplification techniques coupled to a smartphone app and sensor to collect and disseminate acquired data, providing new types of useful data for infectious disease monitoring, modeling, and prediction. Kit will be widely deployed, including to communities and schools, for vector surveillance, public engagement, and educational purposes.

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The role of telemedicine in response to the Zika virus outbreak in Brazil

The International Severe Acute Respiratory and emerging Infection Consortium (ISARIC) has been working with partners in support of the research response to the ZIKV outbreak

A modelling analysis of Zika transmission dynamics and associated microcephaly risk in Brazil

Zika virus induces cell death in human iPSC derived neuronal cells

Zika Virus Infection with Prolonged Maternal Viremia and Fetal Brain Abnormalities - Virology findings

Biology of Zika virus infection in human skin cells

Zika virus: the African and the Asian-Pacific-American lineages

Combination of the sterile insect technique (SIT) with Wolbachia-based approaches for a safe and sustainable solution to control zika vectors

Phylogeography and molecular evolution of Zika Virus

Point-of-care quantitative diagnostics of Dengue and Zika using magnetic nanoparticles

Highly specific serodiagnosis of acute and past Zika virus infections by NSl-based ELISA

Historical Perspectives on Aedes Aegypti Suppression and Eradication Efforts in the Americas

Microcephaly and Zika infection: Preliminary Report of a Case-Control Study

Introduction and Emergence of Zika virus in French Guiana

The viral polymerase inhibitor 7-deaza-2’-C-methyladenosine inhibits in vitro Zika virus replication and delays disease progression in a robust mouse infection model

Experimental studies of susceptibility of Italian Aedes albopictus to Zika Virus

A novel contamination device that targets multiple life-stages of Aedes vectors

Zika virus infection causes microcephaly-like effects in iPSC-derived human brain organoids

A biosilica-based detection device for (re-) emerging arboviral infectious diseases

Real time RT-PCR for detection of ZIKA virus

Development of Zika virus reverse genetics, vaccine candidates and antibodies

ZIKAVax project: the exosome technology as promising platform for the development of a candidate vaccine against Zika virus

Development of rapid and sensitive ELISA kits for Zika Antibodies (Envelop, NS1, prM, and Capsid) in animals and humans.

Zika virus is a candidate for application of nanofibre based system for noninvasive sublingual immunisation: new technology platform for printed vaccines

Enhancing Zika virus Preparedness and Response in New Orleans, Louisiana, USA.

Surveillance and Container Assessments in Residential Environments for the Prevention of Mosquito-Borne Viruses in New Orleans, Louisiana

The use of copepods as biological control for Aedes aegypti

Imported Zika Virus Outbreak in French Guiana: news insights on epidemiology and diagnosis

Evaluating aerial ultra-low volume (ULV) adulticiding applications for Aedes spp. control

Descriptive and prospective study of Zika virus disease in Army forces in French Guiana (ZIFAG)

In-depth analysis of the human centrosome proteome reveals novel factors associated with innate immune signaling and development

Exploration of a study design to estimate male-to-female sexual transmission of Zika virus

Preparing Mexico for the Zika epidemic

Neural stem cell infection by Zika virus in the mouse developing neocortex

The highly structured flaviviral 3’UTR determines transmission of flaviviruses by mosquitoes

Emerging mosquito-borne viruses threats to Europe: Entomological preparedness and risk assessment for emerging arboviral diseases in Greece including Zika, dengue and chikungunya

Miniaturized peptide based differential serology

Isolation of infective Zika virus from urine and saliva of patients in Brazil

Full-length genome sequencing and analysis of 3 ZIKV strains on an Ion Torrent PGM Sequencer

ZIKA VIRUS: The onset in Brazil

Ethico-legal and societal issues and implications of emerging Zika virus epidemics complications and measures

Epitope analysis of Zika virus