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

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

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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 te01-4147c 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 fume mini flavors 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.