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Article

Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response

by
Alexey A. Lozhkov
1,2,
Marina A. Plotnikova
2,
Marya A. Egorova
2,
Irina L. Baranovskaya
1,2,
Ekaterina A. Elpaeva
2,
Sergey A. Klotchenko
1,2,* and
Andrey V. Vasin
1,2,3
1
Institute of Biomedical Systems and Biotechnologies, Peter the Great Saint Petersburg Polytechnic University, 195251 St. Petersburg, Russia
2
Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia
3
Scientific and Educational Center for Biophysical Research in the Field of Pharmaceuticals, Saint Petersburg State Chemical Pharmaceutical University, 197022 St. Petersburg, Russia
*
Author to whom correspondence should be addressed.
Viruses 2022, 14(10), 2090; https://doi.org/10.3390/v14102090
Submission received: 29 July 2022 / Revised: 15 September 2022 / Accepted: 16 September 2022 / Published: 21 September 2022
(This article belongs to the Special Issue Viruses Research in Russia 2022)

Abstract

In this study, we developed a novel, multiplex qPCR assay for simultaneous detection of RIG-1, MDA5, and IFIT-1 at the mRNA level. The assay was validated in A549 cells transfected with in vitro transcribed RNAs. Both exogenous RNA-GFP and self-amplifying (saRNA-GFP) induced significant expression of RIG-1, MDA5, IFIT-1, as well as type I and III interferons. In contrast, native RNA from intact A549 cells did not upregulate expression of these genes. Next, we evaluated RIG-1, MDA5, and IFIT-1 mRNA levels in the white blood cells of patients with influenza A virus (H3N2) or SARS-CoV-2. In acute phase (about 4 days after disease onset) both viruses induced these genes expression. Clinical observations of SARS-CoV-2 typically describe a two-step disease progression, starting with a mild-to-moderate presentation followed by a secondary respiratory worsening 9 to 12 days after the first onset of symptoms. It revealed that the expression of RIG-1, MDA5, and MxA was not increased after 2 and 3 weeks from the onset the disease, while for IFIT-1 it was observed the second peak at 21 day post infection. It is well known that RIG-1, MDA5, and IFIT-1 expression is induced by the action of interferons. Due to the ability of SOCS-1 to inhibit interferon-dependent signaling, and the distinct antagonism of SARS-CoV-2 in relation to interferon-stimulated genes expression, we assessed SOCS-1 mRNA levels in white blood cells. SARS-CoV-2 patients had increased SOCS-1 expression, while the influenza-infected group did not differ from heathy donors. Moreover, SOCS-1 mRNA expression remained stably elevated during the course of the disease. It can be assumed that augmented SOCS-1 expression is one of multiple mechanisms that allow SARS-CoV-2 to escape from the interferon-mediated immune response. Our results implicate SOCS-1 involvement in the pathogenesis of SARS-CoV-2.
Keywords: qPCR; cytosolic sensors; interferons; ISGs; in vitro transcribed RNAs; influenza A virus; SARS-CoV-2; SOCS-1 qPCR; cytosolic sensors; interferons; ISGs; in vitro transcribed RNAs; influenza A virus; SARS-CoV-2; SOCS-1

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MDPI and ACS Style

Lozhkov, A.A.; Plotnikova, M.A.; Egorova, M.A.; Baranovskaya, I.L.; Elpaeva, E.A.; Klotchenko, S.A.; Vasin, A.V. Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response. Viruses 2022, 14, 2090. https://doi.org/10.3390/v14102090

AMA Style

Lozhkov AA, Plotnikova MA, Egorova MA, Baranovskaya IL, Elpaeva EA, Klotchenko SA, Vasin AV. Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response. Viruses. 2022; 14(10):2090. https://doi.org/10.3390/v14102090

Chicago/Turabian Style

Lozhkov, Alexey A., Marina A. Plotnikova, Marya A. Egorova, Irina L. Baranovskaya, Ekaterina A. Elpaeva, Sergey A. Klotchenko, and Andrey V. Vasin. 2022. "Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response" Viruses 14, no. 10: 2090. https://doi.org/10.3390/v14102090

APA Style

Lozhkov, A. A., Plotnikova, M. A., Egorova, M. A., Baranovskaya, I. L., Elpaeva, E. A., Klotchenko, S. A., & Vasin, A. V. (2022). Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response. Viruses, 14(10), 2090. https://doi.org/10.3390/v14102090

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