Comparative analysis of MxA, OAS1, PKR gene expression levels in leukocytes of patients with influenza and coronavirus infection

BACKGROUND: The innate immune response, particularly the interferon system, plays a crucial role in defending the host against viral pathogens. Interferon signaling induces the expression of specific antiviral proteins known as interferon-stimulated genes, which inhibit viral replication through various mechanisms. AIM: This study aimed to develop a quantitative PCR system to assess the molecular regulation of human interferon-stimulated genes MxA, OAS1, and PKR, and to determine their expression in blood leukocytes in response to RNA-containing viruses. MATERIALS AND METHODS: Leukocytes were isolated from patients with laboratory-confirmed influenza and COVID-19 infections 3–4 days after symptom onset. Ex vivo viral infection was induced using influenza viruses A/California/07/09pdm (H1N1pdm09), B/Malaysia/2506/04 (Vic), strain A2 respiratory syncytial virus, and SARS-CoV-2 HCoV-19/Russia/SPE-RII-3524V/2020. RESULTS: A multiplex qPCR assay was developed for analyzing human MxA, OAS1, and PKR gene expression, with high amplification efficiency. The test system was used to study the molecular regulation of these genes in leukocytes in influenza and COVID-19 patients. The expression levels of MxA, OAS1, and PKR genes were significantly increased in blood leukocytes of hospitalized patients 3–4 days after symptom onset. Stimulation of leukocytes by influenza A, influenza B, and respiratory syncytial virus led to increased mRNA levels of these genes, while stimulation by SARS-CoV-2 did not result in changes in gene expression. CONCLUSIONS: The multiplex test system can be used to characterize the expression of antiviral effector interferon-stimulated genes, aiding in the study of virus evasion from the innate immune response.