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Despite in vitro activity of interferon-β (IFN-β) against SARS-CoV-2 infection, its clinical efficacy remains controversial. We evaluated the impact of IFN-β treatment in a cohort of 3590 patients hospitalized with COVID-19 during March–April 2020. The primary endpoint was a composed variable of admission to intensive care unit (ICU)/death. Overall, 153 patients (4%) received IFN-β. They were significantly more severely ill, with a worse clinical and analytical situation, explaining a higher ICU admission (30% vs. 17%; p < 0.01), and a shorter time to the composed variable. In a Cox regression analysis, older age, lymphopenia, renal failure, or increased neutrophil-to-lymphocyte ratio were associated with a greater hazard ratio (HR) of admission at ICU/death. Notably, the HR of IFN-β for the outcome variable was no longer significant after adjustment (HR, 1.03; 95% CI, 0.82–1.30), and different sensitivity analysis (early IFN use, ICU admission) showed no changes in the estimates. A propensity score matching analysis showed no association of IFN-β therapy and outcome. In conclusion, in this large cohort of hospitalized COVID-19 patients, IFN-β was used mainly in patients with advanced disease, reflecting an important bias of selection. After adjusting by severity, IFN-β was not associated with a higher rate of ICU admission or mortality. [ABSTRACT FROM AUTHOR]