Modeling of mixed infection with Zika and West Nile viruses (Flaviviridae: Orthoflavivirus: Orthoflavivirus zikaense, Orthoflavivirus nilense) in vitro and in vivo

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Abstract

Introduction. Mosquito-borne human diseases caused by Zika virus and West Nile virus (WNV) are widespread across multiple continents and cause major outbreaks. Their ranges overlap and the possibility of mixed infections is obvious. The information of such mixed infections is limited.

The aim of the study is to investigate the features of mixed infection of WNV and Zika virus in vitro and in vivo in order to assess their possible interference and/or enhancement of viral infection.

Materials and methods. The study used West Nile virus and Zika virus strains Vlg27924 and MR766, respectively. The infectious activity of viruses during mono- and co-infection was determined on Vero E6 cell culture using RT-PCR, as well as on BALB/c mice using various administration schemes.

Results. In vitro studies of co-infection with WNV and Zika virus showed that co-infection leads to interference, with the degree of competitive inhibition of replication being more pronounced for Zika virus, reaching 1000 times or more when compared to mono-infection. During simultaneous infection in mice, Zika virus does not affect the development of lethal infection caused by WNV. However, preliminary (4 and 20 days) infection with a sublethal dose of Zika virus reliably protects animals from subsequent administration of 10 and 100 LD50 WNV, respectively. In pre-infected and co-infected animals with Zika virus, the development of WNV-specific viral neutralizing antibodies was recorded in higher titers than in WNV monoinfected animals.

Conclusion. The presence of in vitro interference between the studied orthoflaviviruses was shown, most pronounced in relation to the Zika virus. No significant effect was observed with simultaneous co-infection in vivo. However, pre-infection of mice with Zika virus provides protection to animals from lethal WNV infection due to the induction of high levels of antibodies that specifically neutralize its infectious activity.

About the authors

Victor A. Svyatchenko

State Research Center of Virology and Biotechnology «Vector»

Email: svyat@vector.nsc.ru
ORCID iD: 0000-0002-2729-0592

Candidate of Biological Sciences, Leading Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Elena V. Protopopova

State Research Center of Virology and Biotechnology «Vector»

Email: protopopova_ev@vector.nsc.ru
ORCID iD: 0000-0002-2782-8364

Candidate of Biological Sciences, Leading Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Stanislav S. Legostaev

State Research Center of Virology and Biotechnology «Vector»

Email: legostaev_ss@vector.nsc.ru
ORCID iD: 0000-0002-6202-445X

junior researcher of the department of molecular virology of flaviviruses and viral hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Tamara P. Mikryukova

State Research Center of Virology and Biotechnology «Vector»

Email: mikryukova_tp@vector.nsc.ru
ORCID iD: 0000-0003-4350-4260

Candidate of Biological Sciences, Senior Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Alexander P. Agafonov

State Research Center of Virology and Biotechnology «Vector»

Email: agafonov@vector.nsc.ru
ORCID iD: 0000-0003-2577-0434

Doctor of Biological Sciences, Director General

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Valery B. Loktev

State Research Center of Virology and Biotechnology «Vector»

Author for correspondence.
Email: loktev@vector.nsc.ru
ORCID iD: 0000-0002-0229-321X

MD, PhD, DSc, Prof., academician RANS, Chief Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk Region

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3. Fig. 1. Survival of BALB/c mice infected with West Nile virus (WNV) and Zika virus under different schemes of virus administration.  – experimental results obtained by intraperitoneal Zika virus infection with a dose of 104 TCID50; ● – experimental results obtained by intraperitoneal infection of WNV with a dose of 103 TCID50 (10 LD50); □ – experimental results obtained by simultaneous intraperitoneal infection with Zika virus (104 TCID50) and WNV (10 LD50); ■ – experimental results obtained by intraperitoneal infection with Zika virus (104 TCID50) and after 4 days intraperitoneal infection with 10 LD50 of WNV (n = 10; p < 0.05).

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4. Fig. 2. Survival of Zika virus pre-infected BALB/c mice upon subsequent infection with lethal doses of West Nile virus (WNV).  – experimental results obtained by intraperitoneal infection with Zika virus (104 TCID50) and after 20 days intraperitoneal infection with WNV (10 LD50); ● – experimental results obtained by intraperitoneal infection with WNV (100 LD50); □ – experimental results obtained with intraperitoneal infection of WNV (10 LD50); ■ – experimental results obtained by intraperitoneal infection with Zika virus (104 TCID50) and after 20 days intraperitoneal infection with WNV (100 LD50) (n = 10; p < 0.05).

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Copyright (c) 2025 Svyatchenko V.A., Protopopova E.V., Legostaev S.S., Mikryukova T.P., Agafonov A.P., Loktev V.B.

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