Elevated plasma levels of the long pentraxin, pentraxin 3, in severe dengue virus infections

ABSTRACT Dengue virus infections, which have been reported in nearly 140 countries, pose a significant threat to human health. The genome of dengue virus encodes three structural and seven nonstructural (NS) proteins along with two untranslated regions, one each on both ends. Among them, dengue protease (NS3) plays a pivotal role in polyprotein processing and virus multiplication. NS3 is also known to regulate several host proteins to induce and maintain pathogenesis. Certain viral proteins are known to interact with mitochondrial membrane proteins and interfere with their functions, but the association of a virus-coded protein with the mitochondrial matrix is not known. In this report, by using in silico analysis, we show that NS3pro alone is capable of mitochondrial import; however, this is dependent on its innate mitochondrial transport signal (MTS). Transient-transfection and protein import studies confirm the import of NS3pro to the mitochondrial matrix. Similarly, NS3pro-helicase (amino acids 1 to 464 of NS3) also targets the mitochondria. Intriguingly, reduced levels of matrix-localized GrpE protein homolog 1 (GrpEL1), a cochaperone of mitochondrial Hsp70 (mtHsp70), were noticed in NS3pro-expressing, NS3pro-helicase-expressing, and virus-infected cells. Upon the use of purified components, GrpEL1 undergoes cleavage, and the cleavage sites have been mapped to KR81A and QR92S. Importantly, GrpEL1 levels are seriously compromised in severe dengue virus-infected clinical samples. Our studies provide novel insights into the import of NS3 into host mitochondria and identify a hitherto unknown factor, GrpEL1, as a cleavage target, thereby providing new avenues for dengue virus research and the design of potential therapeutics. IMPORTANCE Approximately 40% of the world’s population is at risk of dengue virus infection. There is currently no specific drug or potential vaccine for these infections. Lack of complete understanding of the pathogenesis of the virus is one of the hurdles that must be overcome in developing antivirals for this virus infection. In the present study, we observed that the dengue virus-coded protease imports to the mitochondrial matrix, and our report is the first ever of a virus-coded protein, either animal or human, importing to the mitochondrial matrix. Our analysis indicates that the observed mitochondrial import is due to an inherited mitochondrial transport signal. We also show that matrix-localized GrpE protein homolog 1 (GrpEL1), a cochaperone of mitochondrial Hsp70 (mtHsp70), is also the substrate of dengue virus protease, as observed in vitro and ex vivo in virus-infected cells and dengue virus-infected clinical samples. Hence, our studies reveal an essential aspect of the pathogenesis of dengue virus infections, which may aid in developing antidengue therapeutics.

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High plasma mid-regional pro-adrenomedullin levels in children with severe dengue virus infections

Journal of Clinical Virology ◽

10.1016/j.jcv.2010.09.008 ◽

2011 ◽

Vol 50 (1) ◽

pp. 8-12 ◽

Cited By ~ 9

Author(s):

Meta Michels ◽

Kis Djamiatun ◽

Sultana M.H. Faradz ◽

Mieke M.J.F. Koenders ◽

Quirijn de Mast ◽

...

Keyword(s):

Dengue Virus ◽

High Plasma ◽

Severe Dengue ◽

Virus Infections

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Adapting Rapid Diagnostic Tests to Detect Historical Dengue Virus Infections

Frontiers in Immunology ◽

10.3389/fimmu.2021.703887 ◽

2021 ◽

Vol 12 ◽

Author(s):

Fernando Echegaray ◽

Peter Laing ◽

Samantha Hernandez ◽

Sully Marquez ◽

Amanda Harris ◽

...

Keyword(s):

Dengue Virus ◽

Diagnostic Tests ◽

High Specificity ◽

Denv Infection ◽

Cross Reactivity ◽

Rapid Diagnostic Tests ◽

World Health ◽

Severe Dengue ◽

Virus Infections ◽

Health Organization

The only licensed dengue vaccine, Dengvaxia®, increases risk of severe dengue when given to individuals without prior dengue virus (DENV) infection but is protective against future disease in those with prior DENV immunity. The World Health Organization has recommended using rapid diagnostic tests (RDT) to determine history of prior DENV infection and suitability for vaccination. Dengue experts recommend that these assays be highly specific (≥98%) to avoid erroneously vaccinating individuals without prior DENV infection, as well as be sensitive enough (≥95%) to detect individuals with a single prior DENV infection. We evaluated one existing and two newly developed anti-flavivirus RDTs using samples collected >6 months post-infection from individuals in non-endemic and DENV and ZIKV endemic areas. We first evaluated the IgG component of the SD BIOLINE Dengue IgG/IgM RDT, which was developed to assist in confirming acute/recent DENV infections (n=93 samples). When evaluated following the manufacturer’s instructions, the SD BIOLINE Dengue RDT had 100% specificity for both non-endemic and endemic samples but low sensitivity for detecting DENV seropositivity (0% non-endemic, 41% endemic). Sensitivity increased (53% non-endemic, 98% endemic) when tests were allowed to run beyond manufacturer recommendations (0.5 up to 3 hours), but specificity decreased in endemic samples (36%). When tests were evaluated using a quantitative reader, optimal specificity could be achieved (≥98%) while still retaining sensitivity at earlier timepoints in non-endemic (44-88%) and endemic samples (31-55%). We next evaluated novel dengue and Zika RDTs developed by Excivion to detect prior DENV or ZIKV infections and reduce cross-flavivirus reactivity (n=207 samples). When evaluated visually, the Excivion Dengue RDT had sensitivity and specificity values of 79%, but when evaluated with a quantitative reader, optimal specificity could be achieved (≥98%) while still maintaining moderate sensitivity (48-75%). The Excivion Zika RDT had high specificity (>98%) and sensitivity (>93%) when evaluated quantitatively, suggesting it may be used alongside dengue RDTs to minimize misclassification due to cross-reactivity. Our findings demonstrate the potential of RDTs to be used for dengue pre-vaccination screening to reduce vaccine-induced priming for severe dengue and show how assay design adaptations as well quantitative evaluation can further improve RDTs for this purpose.

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Vascular Leakage in Severe Dengue Virus Infections: A Potential Role for the Nonstructural Viral Protein NS1 and Complement

The Journal of Infectious Diseases ◽

10.1086/500949 ◽

2006 ◽

Vol 193 (8) ◽

pp. 1078-1088 ◽

Cited By ~ 302

Author(s):

Panisadee Avirutnan ◽

Nuntaya Punyadee ◽

Sansanee Noisakran ◽

Chulaluk Komoltri ◽

Somchai Thiemmeca ◽

...

Keyword(s):

Dengue Virus ◽

Viral Protein ◽

Potential Role ◽

Vascular Leakage ◽

Severe Dengue ◽

Virus Infections

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Differential Gene Expression Changes in Children with Severe Dengue Virus Infections

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0000215 ◽

2008 ◽

Vol 2 (4) ◽

pp. e215 ◽

Cited By ~ 34

Author(s):

Martijn D. de Kruif ◽

Tatty E. Setiati ◽

Albertus T. A. Mairuhu ◽

Penelopie Koraka ◽

Hella A. Aberson ◽

...

Keyword(s):

Gene Expression ◽

Dengue Virus ◽

Differential Gene Expression ◽

Severe Dengue ◽

Virus Infections ◽

Differential Gene

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Co-circulation and co-infections of all dengue virus serotypes in Hyderabad, India 2014

Epidemiology and Infection ◽

10.1017/s0950268817001479 ◽

2017 ◽

Vol 145 (12) ◽

pp. 2563-2574 ◽

Cited By ~ 16

Author(s):

K. VADDADI ◽

C. GANDIKOTA ◽

P. K. JAIN ◽

V. S. V. PRASAD ◽

M. VENKATARAMANA

Keyword(s):

Dengue Virus ◽

Severe Dengue ◽

Nucleotide Sequencing ◽

Virus Infections ◽

Disease Epidemiology ◽

Igm Elisa ◽

Dengue Virus Serotypes ◽

First Time ◽

Close Relatedness

SUMMARYThe burden of dengue virus infections increased globally during recent years. Though India is considered as dengue hyper-endemic country, limited data are available on disease epidemiology. The present study includes molecular characterization of dengue virus strains occurred in Hyderabad, India, during the year 2014. A total of 120 febrile cases were recruited for this study, which includes only children and 41 were serologically confirmed for dengue positive infections using non-structural (NS1) and/or IgG/IgM ELISA tests. RT-PCR, nucleotide sequencing and evolutionary analyses were carried out to identify the circulating serotypes/genotypes. The data indicated a high percent of severe dengue (63%) in primary infections. Simultaneous circulation of all four serotypes and co-infections were observed for the first time in Hyderabad, India. In total, 15 patients were co-infected with more than one dengue serotype and 12 (80%) of them had severe dengue. One of the striking findings of the present study is the identification of serotype Den-1 as the first report from this region and this strain showed close relatedness to the Thailand 1980 strains but not to any of the strains reported from India until now. Phylogenetically, all four strains of the present study showed close relatedness to the strains, which are reported to be high virulent.

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