key: cord-0787738-cnydxkjo authors: Akbari, Mohammadarian; Akhavan-Bahabadi, Mehdi; Shafigh, Navid; Taheriazam, Afshin; Hussen, Bashdar Mahmud; Sayad, Arezou; Fathi, Mohadeseh; Taheri, Mohammad; Ghafouri-Fard, Soudeh; Fathi, Mohammad title: Expression analysis of IFNAR1 and TYK2 transcripts in COVID-19 patients date: 2022-03-04 journal: Cytokine DOI: 10.1016/j.cyto.2022.155849 sha: 620f4f49677a2bd4015f9bb633622a285a275e8d doc_id: 787738 cord_uid: cnydxkjo As a member of JAK family of non-receptor tyrosine kinases, TYK2 has a crucial role in regulation of immune responses. This protein has a crucial role in constant expression of IFNAR1 on surface of cells and initiation of type I IFN signaling. In the current study, we measured expression of IFNAR1 and TYK2 levels in venous blood samples of COVID-19 patients and matched controls. TYK2 was significantly down-regulated in male patients compared with male controls (RME=0.34, P value=0.03). Though, levels of TYK2 were not different between female cases and female controls, or between ICU-admitted and non-ICU-admitted cases. Expression of IFNAR1 was not different either between COVID-19 cases and controls or between patients required ICU admission and non-ICU-admitted cases. However, none of these transcripts can properly diffrentiate COVID-19 cases from controls or separate patients based on disease severity. The current study proposes down-regulation of TYK2 as a molecular mechanism for incapacity of SARS-CoV-2 in induction of a competent IFN response. Tyrosine kinase 2 (TYK2) gene encodes a member of the Janus kinase (JAK) family of nonreceptor tyrosine kinases. These kinases have important roles in the regulation of immune response and cell development (1) . This protein has functional association with IFNAR1 receptor subunit. This association has a positive influence on ligand binding to the receptor complex. In fact, TYK2 has a crucial role in stable expression of IFNAR1 on cell surface (2) . Thus, proper activity of TYK2 is a crucial step for initiation of type I IFN response (3) . IFNs are important antiviral cytokines that diminish the impacts of attacking viruses during early phase of viral infections (4) . Recent studies have shown inability of COVID-19 infection in induction of a competent IFN response to decrease the severity of the viral infection (4, 5) . Although several mechanisms might be involved in this process, an imperfect function of IRF3 in activation of the IFN-β promoter has been suggested as a possible mechanism for incapacity of SARS-CoV-2 in induction of a competent IFN response (4, 6) . Another study has revealed the presence of potential inactivating variants in genes related with Toll-like receptors the type I IFN pathway in a proportion of severely affected COVID-19 patients, emphasizing on the importance of these pathways in protection against severe disorder (7) . Consistent with this finding, autoantibodies against type I IFN have been detected in a number of severely affected COVID-19 patients. Notably, most of these autoantibodies had neutralising ability in vitro (8) . Although the importance of type I IFN responses has been well established in defence against SARS-CoV-2 and related viral infections, the mechansim of such malfunctioning has not been completely understood. In the current study, we measured expression of IFNAR1 and TYK2 levels in venous blood samples of COVID-19 patients and matched controls to unravel their role in determination of the course of COVID-19. The current study was performed on 91 COVID-19 cases admitted to Nikan Hospital, Tehran, during 2020. Diagnosis was confirmed through assessment of nasopharyngeal swab samples. Four milliliters of venous blood were gathered from all cases and healthy individuals. Next, total RNA was retrieved from blood specimens using the TRIzol reagent. Then, complementary DNA was created from these specimens by using the Smobio cDNA production kit (Taiwan). Transcript quantities of IFNAR1 and TYK2 genes were quantified in all samples using the real time PCR Master Mix (Amplicon, Denmark). Primers are summarized in Table 1 . were considered as significant. Female/male raito was 38/53 and 39/52 in cases and controls, respectively. The mean age patients and non-ICU-admitted cases. TYK2 was significantly down-regulated in male patients compared with male controls (RME=0.34, P value=0.03). Nonetheless, expression of TYK2 was not different between female cases and female controls, or between ICU-admitted and non ICU-admitted cases. Expression of IFNAR1 was not different either between COVID-19 cases and controls or between patients required ICU admission and those did not require ICU admission ( Table 2 ). We also performed a multivariate analysis using linear regression model to assess correlations between expressions of TYK2 and IFNAR1 and clinical variables (Table 3) . IFNAR1 expression levels were significantly correlated with MCHC. Thus, the multivariate analysis showed that that the bivariate correlations presented in Figure 5 are not real. Finally, we depicted ROC curves to assess diagnostic power of IFNAR1 and TYK2 genes in separation of COVID-19 cases from controls as well as patients required ICU admission and those not required ICU admission ( Figures 6A and 6B , respectively). (Table 4 ). protein levels. We also emphasize that these results should be verified in further studies. Tyrosine kinase 2 (TYK2), Janus kinase (JAK), AUC (Area under curve). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent forms were obtained from all study participants. Informed consent forms were obtained from all study participants and from legally authorized representative/next of kin of deceased patients. The study protocol was approved by the ethical committee of Shahid Beheshti University of Medical Sciences (IR.SBMU.RETECH.REC.1399.592). All methods were performed in accordance with the relevant guidelines and regulations. Not applicable The analyzed data sets generated during the study are available from the corresponding author on reasonable request. The authors declare they have no conflict of interest This study was financially supported by Shahid Beheshti University of Medical Sciences. SGF and MT wrote the draft and revised it. MF designed and supervised the study. NS, AT and MF performed the experiment and data collection. AS and MAB analyzed the data. All the authors read and approved the submitted version. 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Arthritis and rheumatism RNA-binding protein RBM47 stabilizes IFNAR1 mRNA to potentiate host antiviral activity Not applicable SGF and MT wrote the draft and revised it. MF designed and supervised the study. NS, AT and MF performed the experiment and data collection. AS and MAB analyzed the data. All the authors read and approved the submitted version. The authors declare they have no conflict of interest.