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: ISGs; SARS-CoV-2; SOCS-1; cytosolic sensors; in vitro transcribed RNAs; influenza A virus; interferons; qPCR.
【저자키워드】 SARS-CoV-2, interferons, influenza A virus, ISGs, qPCR., in vitro transcribed RNAs, cytosolic sensors, SOCS-1, 【초록키워드】 immune response, in vitro, virus, influenza A, RNAs, RNA, Disease progression, White blood cell, MDA5, qPCR, clinical, donors, multiplex, expression, Signaling, disease onset, Mild-to-moderate, observation, White blood cells, acute phase, type I, worsening, H3N2, mRNA expression, qPCR assay, interferon-stimulated gene, post infection, mRNA level, pathogenesis of SARS-CoV-2, Genes, Course, not differ, remained, inhibit, the disease, evaluated, elevated, transcribed, onset of symptoms, cytosolic, both virus, assumed, A549 cell, multiple mechanism, patients with influenza, SARS-CoV-2 patient, transfected with, 【제목키워드】 detection, MDA5, response, Simultaneous,