Asian Zika virus strains target CD14+ blood monocytes and induce M2-skewed immunosuppression during pregnancy

Abstract Background Monitoring both invasion of Zika virus disease into free countries and circulation in endemic countries is essential to avoid a global pandemic. However, the difficulty lies in detecting Zika virus due to the large variety of mutations in its genomic sequence. To develop a rapid and simple method with high accuracy, reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was adopted for the detection of Zika virus strains derived from several countries. Results Common primers for RT-LAMP were designed based on the genomic sequences of two standard Zika strains: African lineage, MR-766, and Asian lineage, PRVABC59. RT-LAMP reactions using a screened primer set, targeting the NS3 region, detected both Zika virus strains. The minimum detectable quantity was 3 × 10−2 ng of virus RNA. Measurable lag of reaction times among strains was observed. The RT-LAMP method amplified the target virus sequence from the urine and serum of a patient with a travel history in the Caribbean Islands and also provided a prediction about which lineage of Zika virus strain was present. Conclusions The RT-LAMP method using a well-optimized primer set demonstrated high specificity and sensitivity for the detection of Zika virus strains with a variety in genomic RNA sequences. In combination with the simplicity of LAMP reaction in isothermal conditions, the optimized primer set established in this study may facilitate rapid and accurate diagnosis of Zika fever patients with virus strain information.

Download Full-text

Two Genetic Differences between Closely Related Zika Virus Strains Determine Pathogenic Outcome in Mice

Journal of Virology ◽

10.1128/jvi.00618-20 ◽

2020 ◽

Vol 94 (20) ◽

Cited By ~ 3

Author(s):

Derek L. Carbaugh ◽

Shuntai Zhou ◽

Wes Sanders ◽

Nathaniel J. Moorman ◽

Ronald Swanstrom ◽

...

Keyword(s):

Amino Acid ◽

Mouse Models ◽

Zika Virus ◽

Clinical Manifestations ◽

Amino Acid Sequences ◽

Viral Envelope ◽

Laboratory Research ◽

Balanced Polymorphism ◽

E Protein ◽

Virus Strains

ABSTRACT Recent Zika virus (ZIKV) outbreaks and unexpected clinical manifestations of ZIKV infection have prompted an increase in ZIKV-related research. Here, we identify two strain-specific determinants of ZIKV virulence in mice. We found that strain H/PF/2013 caused 100% lethality in Ifnar1−/− mice, whereas PRVABC59 caused no lethality; both strains caused 100% lethality in Ifnar1−/− Ifngr1−/− double-knockout (DKO) mice. Deep sequencing revealed a high-frequency variant in PRVABC59 not present in H/PF/2013: a G-to-T change at nucleotide 1965 producing a Val-to-Leu substitution at position 330 of the viral envelope (E) protein. We show that the V330 variant is lethal on both virus strain backgrounds, whereas the L330 variant is attenuating only on the PRVABC59 background. These results identify a balanced polymorphism in the E protein that is sufficient to attenuate the PRVABC59 strain but not H/PF/2013. The consensus sequences of H/PF/2013 and PRVABC59 differ by 3 amino acids, but these were not responsible for the difference in virulence between the two strains. H/PF/2013 and PRVABC59 differ by an additional 31 noncoding or silent nucleotide changes. We made a panel of chimeric viruses with identical amino acid sequences but nucleotide sequences derived from H/PF/2013 or PRVABC59. We found that 6 nucleotide differences in the 3′ quarter of the H/PF/2013 genome were sufficient to confer virulence in Ifnar1−/− mice. Altogether, our work identifies a large and previously unreported difference in virulence between two commonly used ZIKV strains, in two widely used mouse models of ZIKV pathogenesis (Ifnar1−/− and Ifnar1−/− Ifngr1−/− DKO mice). IMPORTANCE Contemporary ZIKV strains are closely related and often used interchangeably in laboratory research. Here, we identify two strain-specific determinants of ZIKV virulence that are evident in only Ifnar1−/− mice but not Ifnar1−/− Ifngr1−/− DKO mice. These results identify a balanced polymorphism in the E protein that is sufficient to attenuate the PRVABC59 strain but not H/PF/2013. We further identify a second virulence determinant in the H/PF/2013 strain, which is driven by the viral nucleotide sequence but not the amino acid sequence. Altogether, our work identifies a large and previously unreported difference in virulence between two commonly used ZIKV strains, in two widely used mouse models of ZIKV pathogenesis. Our results highlight that even very closely related virus strains can produce significantly different pathogenic phenotypes in common laboratory models.

Download Full-text

Mutational landscape of Zika virus strains worldwide and its structural impact on proteins

Gene ◽

10.1016/j.gene.2019.05.039 ◽

2019 ◽

Vol 708 ◽

pp. 57-62 ◽

Cited By ~ 1

Author(s):

Almerinda Agrelli ◽

Ronald Rodrigues de Moura ◽

Sergio Crovella ◽

Lucas André Cavalcanti Brandão

Keyword(s):

Zika Virus ◽

Mutational Landscape ◽

Structural Impact ◽

Virus Strains

Download Full-text

Complete Genome Sequences of Three Historically Important, Spatiotemporally Distinct, and Genetically Divergent Strains of Zika Virus: MR-766, P6-740, and PRVABC-59: TABLE 1

Genome Announcements ◽

10.1128/genomea.00800-16 ◽

2016 ◽

Vol 4 (4) ◽

Cited By ~ 19

Author(s):

Sang-Im Yun ◽

Byung-Hak Song ◽

Jordan C. Frank ◽

Justin G. Julander ◽

Irina A. Polejaeva ◽

...

Keyword(s):

Puerto Rico ◽

Zika Virus ◽

Complete Genome ◽

Reverse Genetics ◽

Genomic Sequences ◽

Genome Sequences ◽

African Lineage ◽

American Strain ◽

Asian Lineage ◽

Virus Strains

Here, we report the 10,807-nucleotide-long consensus RNA genome sequences of three spatiotemporally distinct and genetically divergent Zika virus strains, with the functionality of their genomic sequences substantiated by reverse genetics: MR-766 (African lineage, Uganda, 1947), P6-740 (Asian lineage, Malaysia, 1966), and PRVABC-59 (Asian lineage-derived American strain, Puerto Rico, 2015).

Download Full-text