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Simultaneous Detection of RIG-1, MDA5, and IFIT-1 Expression Is a Convenient Tool for Evaluation of the Interferon-Mediated Response.
Lozhkov, Alexey A; Plotnikova, Marina A; Egorova, Marya A; Baranovskaya, Irina L; Elpaeva, Ekaterina A; Klotchenko, Sergey A; Vasin, Andrey V.
  • Lozhkov AA; Institute of Biomedical Systems and Biotechnologies, Peter the Great Saint Petersburg Polytechnic University, 195251 St. Petersburg, Russia.
  • Plotnikova MA; Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia.
  • Egorova MA; Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia.
  • Baranovskaya IL; Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia.
  • Elpaeva EA; Institute of Biomedical Systems and Biotechnologies, Peter the Great Saint Petersburg Polytechnic University, 195251 St. Petersburg, Russia.
  • Klotchenko SA; Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia.
  • Vasin AV; Smorodintsev Research Institute of Influenza, 197376 St. Petersburg, Russia.
Viruses ; 14(10)2022 09 21.
Article in English | MEDLINE | ID: covidwho-2043983
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Interferons / COVID-19 Type of study: Experimental Studies / Observational study / Prognostic study Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: V14102090

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Interferons / COVID-19 Type of study: Experimental Studies / Observational study / Prognostic study Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: V14102090