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Summary Background Hepatitis C virus (HCV) infection is associated with production of different serum non‐organ‐specific antibodies (NOSA) and risk for developing autoimmune disorders. The clinical significance of these phenomena is not fully understood. Aim To assess non‐organ‐specific antibodies before and 24 weeks after the end of therapy with direct‐acting antivirals in patients with HCV‐related infection, to better clarify the clinical relevance of these antibodies in terms of treatment response and prognostic value. Methods Patients enrolled (191) were considered non‐organ‐specific antibody‐positive for titres ≥1:40 on at least two determinations before treatment. Results At baseline, 46 patients were positive and 145 were negative for autoantibodies. The prevalence of autoimmune thyroiditis was significantly higher in non‐organ‐specific antibody‐positive group than non‐organ‐specific antibody‐negative group (P = 0.02). HCV‐RNA 24 weeks after the end of antiviral therapy was 100% negative in patients with antibodies positivity and 98.6% in antibody‐negative patients (P = 1.0). In the former group, autoantibodies disappeared in 30 of 46 patients (65.2%). On multivariate analysis, non‐organ‐specific antibody‐negativity was significantly reduced in patients with hepatic hilar lymphadenopathy (OR = 0.17; 95% CI 0.02‐0.94, P = 0.04). None of the adverse events occurring during antiviral therapy was related to autoimmune disorders. Conclusions Hepatitis C virus clearance frequently reduces non‐organ‐specific antibody positivity suggesting that they represent an epiphenomenon of the viral infection. However, in patients who did not become negative, long‐term monitoring would establish whether they could hide an underlying process that may progress into a clear autoimmune or rheumatologic disease. (Trial registration number: NCT03566966).

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