#Virus-inclusive single-cell RNA #sequencing reveals the molecular signature of #progression to severe #dengue (Proc Natl Acad Sci USA, abstract)

[Source: Proceedings of the National Academy of Sciences of the United States of America, full page: (LINK). Abstract, edited.]

Virus-inclusive single-cell RNA sequencing reveals the molecular signature of progression to severe dengue

Fabio Zanini, Makeda L. Robinson, Derek Croote, Malaya Kumar Sahoo, Ana Maria Sanz, Eliana Ortiz-Lasso, Ludwig Luis Albornoz, Fernando Rosso, Jose G. Montoya, Leslie Goo, Benjamin A. Pinsky, Stephen R. Quake, and Shirit Einav

PNAS published ahead of print December 7, 2018 / DOI: https://doi.org/10.1073/pnas.1813819115

Contributed by Stephen R. Quake, October 24, 2018 (sent for review August 10, 2018; reviewed by Katja Fink and Alex K. Shalek)

 

Significance

A fraction of the 400 million people infected with dengue annually progresses to severe dengue (SD). Yet, there are currently no biomarkers to predict disease progression. We profiled the landscape of host transcripts and viral RNA in thousands of single blood cells from dengue patients prior to progressing to SD. We discovered cell type-specific immune activation and candidate predictive biomarkers. We also determined preferential virus association with specific cell populations, particularly naive B cells and monocytes. We explored immune activation of bystander cells, clonality and somatic evolution of adaptive immune repertoires, as well as viral genomics. This multifaceted approach could advance understanding of pathogenesis of any viral infection, map an atlas of infected cells, and promote the development of prognostics.

 

Abstract

Dengue virus (DENV) infection can result in severe complications. However, the understanding of the molecular correlates of severity is limited, partly due to difficulties in defining the peripheral blood mononuclear cells (PBMCs) that contain DENV RNA in vivo. Accordingly, there are currently no biomarkers predictive of progression to severe dengue (SD). Bulk transcriptomics data are difficult to interpret because blood consists of multiple cell types that may react differently to infection. Here, we applied virus-inclusive single-cell RNA-seq approach (viscRNA-Seq) to profile transcriptomes of thousands of single PBMCs derived early in the course of disease from six dengue patients and four healthy controls and to characterize distinct leukocyte subtypes that harbor viral RNA (vRNA). Multiple IFN response genes, particularly MX2 in naive B cells and CD163 in CD14+ CD16+ monocytes, were up-regulated in a cell-specific manner before progression to SD. The majority of vRNA-containing cells in the blood of two patients who progressed to SD were naive IgM B cells expressing the CD69 and CXCR4 receptors and various antiviral genes, followed by monocytes. Bystander, non-vRNA–containing B cells also demonstrated immune activation, and IgG1 plasmablasts from two patients exhibited clonal expansions. Lastly, assembly of the DENV genome sequence revealed diversity at unexpected sites. This study presents a multifaceted molecular elucidation of natural dengue infection in humans with implications for any tissue and viral infection and proposes candidate biomarkers for prediction of SD.

dengue – single cell – transcriptomics – biomarkers – virus–host interactions

Keywords: Dengue fever; Viral pathogenesis.

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#Sequential #Infection of #Aedes aegypti #Mosquitoes with #Chikungunya Virus and #Zika Virus Enhances Early Zika Virus Transmission (Insects, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Insects. 2018 Dec 1;9(4). pii: E177. doi: 10.3390/insects9040177.

Sequential Infection of Aedes aegypti Mosquitoes with Chikungunya Virus and Zika Virus Enhances Early Zika Virus Transmission.

Magalhaes T1, Robison A2, Young MC3, Black WC 4th4, Foy BD5, Ebel GD6, Rückert C7.

Author information: 1 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. Tereza.Magalhaes@colostate.edu. 2 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. lexir5394@gmail.com. 3 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. emceeyoung@gmail.com. 4 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. William.Black@colostate.edu. 5 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. Brian.Foy@colostate.edu. 6 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. Gregory.Ebel@colostate.edu. 7 Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. Claudia.Rueckert@Colostate.edu.

 

Abstract

In urban settings, chikungunya, Zika, and dengue viruses are transmitted by Aedes aegypti mosquitoes. Since these viruses co-circulate in several regions, coinfection in humans and vectors may occur, and human coinfections have been frequently reported. Yet, little is known about the molecular aspects of virus interactions within hosts and how they contribute to arbovirus transmission dynamics. We have previously shown that Aedes aegypti exposed to chikungunya and Zika viruses in the same blood meal can become coinfected and transmit both viruses simultaneously. However, mosquitoes may also become coinfected by multiple, sequential feeds on single infected hosts. Therefore, we tested whether sequential infection with chikungunya and Zika viruses impacts mosquito vector competence. We exposed Ae. aegypti mosquitoes first to one virus and 7 days later to the other virus and compared infection, dissemination, and transmission rates between sequentially and single infected groups. We found that coinfection rates were high after sequential exposure and that mosquitoes were able to co-transmit both viruses. Surprisingly, chikungunya virus coinfection enhanced Zika virus transmission 7 days after the second blood meal. Our data demonstrate heterologous arbovirus synergism within mosquitoes, by unknown mechanisms, leading to enhancement of transmission under certain conditions.

KEYWORDS: Zika; arboviruses; chikungunya; coinfection; mosquitoes; sequential infection

PMID: 30513725 DOI: 10.3390/insects9040177

Keywords: Arbovirus; Chikungunya fever; Zika Virus; Dengue fever; Mosquitoes; Aedes spp.; Aedes aegypti.

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#Seroprevalence of #Dengue and #Zika Virus in #Blood #Donations: A Systematic Review (Transfus Med Rev., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Transfus Med Rev. 2018 Oct 31. pii: S0887-7963(18)30094-4. doi: 10.1016/j.tmrv.2018.10.001. [Epub ahead of print]

Seroprevalence of Dengue and Zika Virus in Blood Donations: A Systematic Review.

Eick SM1, Dale AP2, McKay B3, Lawrence C4, Ebell MH5, Cordero JF6, Welton M7.

Author information: 1 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: eick.steph@gmail.com. 2 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: aperrydale@gmail.com. 3 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: bmckay81@gmail.com. 4 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: crl99301@uga.edu. 5 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: ebell@uga.edu. 6 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: jcordero@uga.edu. 7 Department of Epidemiology and Biostatistics, College of Public Health, University of Georgia, University of Georgia Health Sciences Campus, Athens, GA. Electronic address: Michael.welton@gmail.com.

 

Abstract

The presence of antibodies to Zika virus (ZIKV) and dengue virus (DENV) can be detected in blood donations. Donation-based surveillance provides an alternative strategy to estimate population prevalence by detecting antibodies that are circulating. To estimate population prevalence, we conducted a systematic review of literature on the seroprevalence of ZIKV and DENV antibodies in blood donations. We searched PubMed and Web of Science for studies that reported the seroprevalence of ZIKV and DENV in blood donations. The title and abstract of each study were screened by 2 reviewers simultaneously for possible inclusion, and the full text of selected studies was reviewed to ensure that they met inclusion criteria (used primary data collection, reported evidence of immunoglobulin M (IgM) or immunoglobulin G (IgG) antibodies in the blood supply, and included a representative sample of the total population). Immunoglobin test measuring levels of antibodies to IgM and IgG and number of positive cases were extracted from each study. No exclusions were made based on language or country. Our initial search identified 1890 studies after excluding duplicates, of which 76 were assessed for full text eligibility to ensure that they met our final inclusion criteria. There were 14 studies included in our review; 11 examined the seroprevalence of DENV, and 3 examined ZIKV. The highest seroprevalence by IgM was 2.82% for DENV and 0.53% for ZIKV. Our results indicate that the seroprevalence of ZIKV and DENV antibody presence in countries with active transmission is higher than reports by traditional surveillance in some countries. This finding is expected due to the large percentage of asymptomatic cases. The highest seroprevalence was observed for IgG, which can persist over long periods of time compared to IgM. Screening of blood donations may help supplement traditional surveillance measures, especially during outbreak settings.

KEYWORDS: Blood donations; Blood supply; Dengue virus; Surveillance; Transmission; Zika virus

PMID: 30471867 DOI: 10.1016/j.tmrv.2018.10.001

Keywords: Zika Virus; Dengue Fever; Blood Safety; Seroprevalence.

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Analysis of #Zika virus neutralizing #antibodies in normal healthy #Thais (Sci Rep., abstract)

[Source: Scientific Reports, full page: (LINK). Abstract, edited.]

Article | OPEN | Published: 21 November 2018

Analysis of Zika virus neutralizing antibodies in normal healthy Thais

Wannapa Sornjai,  Janejira Jaratsittisin,  Prasert Auewarakul,  Nitwara Wikan &  Duncan R. Smith

Scientific Reports, volume 8, Article number: 17193 (2018)

 

Abstract

Zika virus (ZIKV) infections have been reported from all over Thailand, but the number of reported cases remains low, suggesting a degree of immune protection against ZIKV infection. To address this possibility, the presence of ZIKV neutralizing antibodies was determined in serum from 135 healthy Thai adults with a plaque reduction neutralization test (PRNT), and a number of samples were subsequently analyzed for the presence of neutralizing antibodies to dengue virus (DENV) and Japanese encephalitis virus (JEV). Results showed that 70.4% (PRNT50 ≥ 10), 55.6 (PRNT50 ≥ 20) or 22.2% (PRNT90 ≥ 20) of the samples showed neutralizing antibodies to ZIKV. Detailed analysis showed no association between the presence of neutralizing antibodies to other flaviviruses (DENV, JEV) and the presence of ZIKV neutralizing antibodies. These results suggest that the level of ZIKV neutralizing antibodies in the Thai population is enough to dampen the transmission of the virus in Thailand.

Keywords: Zika Virus; Dengue Fever; Seroprevalence; Thailand.

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#Antivirus effectiveness of #ivermectin on #dengue virus type 2 in #Aedes albopictus (PLoS Negl Trop Dis., abstract)

[Source: PLoS Neglected Tropical Diseases, full page: (LINK). Abstract, edited.]

OPEN ACCESS /  PEER-REVIEWED / RESEARCH ARTICLE

Antivirus effectiveness of ivermectin on dengue virus type 2 in Aedes albopictus

Tie-Long Xu, Yin Han, Wei Liu, Xing-Ya Pang, Bin Zheng, Yi Zhang, Xiao-Nong Zhou

Published: November 19, 2018 / DOI: https://doi.org/10.1371/journal.pntd.0006934 / This is an uncorrected proof.

 

Abstract

Background

Dengue fever is the most rapidly spreading mosquito-borne viral disease over the past 50 years, with a 30-fold increase in global incidence. Dengue vector control is a key component for the dengue control strategy, since no absolutely effective vaccine or drug is available yet. However, the rapid rise and spread of mosquito insecticide resistance have become major threats to the efficiency of insecticide-based vector control activities. Thus, innovative vector control tools are badly needed. This study aims to confirm the antivirus effectiveness of ivermectin on dengue virus type 2 (DENV-2) in Aedes albopictus (Skuse, 1894), then to explore its potential use in the combating to the dengue epidemics.

Methods

Aedes albopictus were first infected with DENV-2 in human whole blood, and at the fourth day after infectious blood feeding, they were divided into eight groups. Seven of them were held for six days with access to 0, 2, 4, 8, 16, 32 and 64 ng/ml ivermectin, respectively, and the last one was set as a historical control group, which was stored at -80°C until being detected at the same time with the other groups. Each mosquito was detected using real-time fluorescent RT-PCR kit. DENV-2 RNA concentration (copies/ml) and infection rate in each group were compared.

Results

Both of quantitatively and qualitatively inhibiting effects of ivermectin have been detected in this study. Generally, DENV-2 replicated well in Aedes albopictus without ivermectin intervention, whose virus loads exhibited significantly higher when the mosquitoes were holding from 4 days to 10 days after infectious blood feeding. In contrast, with the treatment of ivermectin, the infection rate was reduced by as much as 49.63%. The regression equation between infection rates (Y2) and ivermectin concentration log2 values (X2) was obtained as Y2 = 91.41–7.21*X2with R2 = 0.89.

Conclusion

Ivermectin can directly or indirectly inhibit DENV-2 multiplication in Aedes albopictus. Moreover, the actual concentration for application in zooprophylaxis needs to be confirmed in the further field trials.

 

Author summary

Dengue fever is one of neglected vector-borne tropical diseases with a 30-fold increase in global incidence recently. In 2012, World Health Organization set a goal to reduce dengue mortality by at least 50% by 2020. Being faced with more challenges in the dengue control programs, such as the increase of dengue outbreaks, lacking absolutely effective vaccine, rise of vector insecticide resistance and so on; innovative vector control tools are urgently needed for current control programs on dengue fever. To find a new avenue in vector control, we for the first time assessed the inhibiting effectiveness of ivermectin on dengue virus type 2 (DENV-2) inside Aedes mosquitoes. We found that about 80% Aedes albopictus mosquitoes were effectively infected with DENV-2 without treatment of ivermectin. But in the groups of ivermectin treatment, the infection rate of DENV-2 and the median of virus loads were significantly reduced by up to 49.63% and 99.99%, respectively. Both quantitatively and qualitatively inhibiting effects of ivermectin were detected. We found out that ivermectin was able to effectively inhibit the DENV-2 multiplication in Aedes albopictus, which may gave us a hint that using ivermectin in some control programs as a zooprophylaxis to block dengue epidemic through inhibiting DENV-2 in field Aedes mosquitoes.

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Citation: Xu T-L, Han Y, Liu W, Pang X-Y, Zheng B, Zhang Y, et al. (2018) Antivirus effectiveness of ivermectin on dengue virus type 2 in Aedes albopictus. PLoS Negl Trop Dis 12(11): e0006934. https://doi.org/10.1371/journal.pntd.0006934

Editor: Waleed Saleh Al-Salem, Saudi Ministry of Health, SAUDI ARABIA

Received: March 17, 2018; Accepted: October 18, 2018; Published: November 19, 2018

Copyright: © 2018 Xu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the paper and its Supporting Information files.

Funding: This project was financially supported by National Key Research and Development Program of China (No. 2016YFC1202000), and International Cooperation Fund of Shanghai Municipality (No. 17430741900). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Keywords: Dengue Fever; Mosquitoes; Aedes albopictus; Ivermectin.

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Cross-Reactive #Dengue Virus #Antibodies Augment #Zika Virus #Infection of #Human #Placental Macrophages (Cell Host Microbe, abstract)

[Source: Cell Host Microbe, full page: (LINK). Abstract, edited.]

Cross-Reactive Dengue Virus Antibodies Augment Zika Virus Infection of Human Placental Macrophages

Matthew G. Zimmerman 8, Kendra M. Quicke 8, Justin T. O’Neal, Nitin Arora, Deepa Machiah, Lalita Priyamvada, Robert C. Kauffman, Emery Register, Oluwaseyi Adekunle, Dominika Swieboda, Erica L. Johnson, Sarah Cordes, Lisa Haddad, Rana Chakraborty, Carolyn B. Coyne, Jens Wrammert, Mehul S. Suthar 9

Published: November 14, 2018 / DOI: https://doi.org/10.1016/j.chom.2018.10.008

 

Highlights

  • Cross-reactive DENV antibodies enhance ZIKV infection in human Hofbauer cells
  • Enhanced ZIKV infection in mid-gestation explants is IgG subclass dependent
  • ZIKV immune complexes target Hofbauer cells within the villous stroma

 

Summary

Zika virus (ZIKV), which emerged in regions endemic to dengue virus (DENV), is vertically transmitted and results in adverse pregnancy outcomes. Antibodies to DENV can cross-react with ZIKV, but whether these antibodies influence ZIKV vertical transmission remains unclear. Here, we find that DENV antibodies increase ZIKV infection of placental macrophages (Hofbauer cells [HCs]) from 10% to over 80% and enhance infection of human placental explants. ZIKV-anti-DENV antibody complexes increase viral binding and entry into HCs but also result in blunted type I interferon, pro-inflammatory cytokine, and antiviral responses. Additionally, ZIKV infection of HCs and human placental explants is enhanced in an immunoglobulin G subclass-dependent manner, and targeting FcRn reduces ZIKV replication in human placental explants. Collectively, these findings support a role for pre-existing DENV antibodies in enhancement of ZIKV infection of select placental cell types and indicate that pre-existing immunity to DENV should be considered when addressing ZIKV vertical transmission.

Keywords: Zika virus – placental macrophages – antibody-dependent enhancement – dengue virus – cross-reactive antibodies – type I interferon – vertical transmission

Keywords: Zika Virus; Dengue Fever; Pregnancy; ADE; Animal models.

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A comparison of #Zika and #dengue #outbreaks using national #surveillance data in the #Dominican Republic (PLoS Negl Trop Dis., abstract)

[Source: PLoS Neglected Tropical Diseases, full page: (LINK). Abstract, edited.]

OPEN ACCESS /  PEER-REVIEWED / RESEARCH ARTICLE

A comparison of Zika and dengue outbreaks using national surveillance data in the Dominican Republic

Leigh R. Bowman , Joacim Rocklöv, Axel Kroeger, Piero Olliaro, Ronald Skewes

Published: November 5, 2018 / DOI: https://doi.org/10.1371/journal.pntd.0006876 / This is an uncorrected proof.

 

Abstract

Background

Aedes-borne arboviruses continue to precipitate epidemics worldwide. In Dominican Republic, the appearance of Zika virus cases that closely followed a large dengue epidemic provided an opportunity to study the different transmission drivers behind these two flaviviruses. Retrospective datasets were used to collect information on the populations at risk and descriptive statistics were used to describe the outbreaks on a national scale.

Methodology/ Principal findings

Expectedly, box plots showed that 75% of dengue was reported in those aged <20 years while Zika infections were more widely dispersed among the population. Dengue attack rates were marginally higher among males at 25.9 per 10,000 population vs. 21.5 per 10,000 population for females. Zika infections appeared to be highly clustered among females (73.8% (95% CI 72.6%, 75.0%; p<0.05)); age-adjusted Zika attack rates among females were 7.64 per 10,000 population compared with 2.72 per 10,000 population among males. R0 calculations stratified by sex also showed a significantly higher metric among females: 1.84 (1.82, 1.87; p<0.05) when compared to males at 1.72 (1.69, 1.75; p<0.05). However, GBS attack rates stratified by sex revealed slightly higher risk in males vs. females, at 0.62 and 0.57 per 10,000 population respectively.

Conclusions/ Significance

Evidence suggests little impact of existing dengue immunity on reported attack rates of Zika at the population level. Confounding of R0 and incident risk calculations by sex-specific over-reporting can alter the reliability of epidemiological metrics, which could be addressed using associated proxy syndromes or conditions to explore seemingly sex-skewed incidence. The findings indicate that community awareness campaigns, through influencing short-term health seeking behaviour, remain the most plausible mechanism behind increased reporting among women of reproductive age, although biological susceptibility cannot yet be ruled out. Media campaigns and screening are therefore recommended for women of reproductive age during Zika outbreaks. Future research should focus on clinical Zika outcomes among dengue seropositive individuals.

 

Author summary

During the years 2013–2016, worldwide Zika epidemics spread quickly throughout immune-naïve human populations. In 2015–2016, the Dominican Republic was struck by an epidemic of dengue followed by an outbreak of Zika. The Zika epidemic closely followed the tail of the dengue epidemic and provided an opportunity to study the dynamics of Zika and dengue transmission at the population level over a similar timeframe. The findings showed that while dengue cases were marginally higher among males, Zika cases were much higher among females and in particular among sexually active age groups. This might indicate that 1) prior dengue infections (and hence antibody cross-reactivity) in these populations had no material impact on the likelihood of reporting Zika illness 2) over-reporting in females was due to health campaigning that targeted females or 3) there was a biological cause for increased susceptibility among women. These possible causes are not mutually exclusive but it is most likely that the clustering of cases among women was the result of health-seeking behaviour caused by health campaigns and media coverage of Zika at the time. If so, media campaigns can be an effective strategy for encouraging health-seeking behaviour among target demographics.

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Citation: Bowman LR, Rocklöv J, Kroeger A, Olliaro P, Skewes R (2018) A comparison of Zika and dengue outbreaks using national surveillance data in the Dominican Republic. PLoS Negl Trop Dis 12(11): e0006876. https://doi.org/10.1371/journal.pntd.0006876

Editor: David Harley, University of Queensland, AUSTRALIA

Received: March 14, 2018; Accepted: September 26, 2018; Published: November 5, 2018

Copyright: © 2018 Bowman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All data files are available from the Open Science Framework database (https://osf.io/VYN2B/).

Funding: This work was performed as part of the Umeå Centre for Microbial Research (UCMR) Linnaeus Program supported by Umeå University and the Swedish Research Council (grant no. 349-2007-8673). LRB received partial funding from WHO-TDR. The funders had no influence on the research conducted. JR received partial funding from the European Union’s Horizon 2020 research and innovation programme under ZikaPLAN grant agreement No 734584. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist. PO is a staff member of the WHO; the authors alone are responsible for the opinion expressed in this paper which may not reflect the options and policies of WHO.

Keywords: Zika Virus; Dengue Fever; Dominican Republic.

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