It is unknown which intervention strategies are used or effective to increase influenza vaccine uptake by healthcare workers (HCWs) in acute hospitals in England. We undertook a survey in acute hospitals, described strategies employed from 2008 to 2012 and used multivariable binomial regression to identify those effective. Eighty out of 166 trusts responded and reported 25 strategies. Every intervention showed increased use: peer vaccination from 3·8% to 38·8% (+921%); educational DVDs from 3·8% to 22·5% (+492%); Twitter from 2·5% to 12·5% (+400%) and Facebook from 1·3% to 6·3% (+384%). Peer vaccination increased uptake by 7·3% [95% confidence interval (CI) 1·1-13·6, P = 0·02] overall; educational DVDs by 9·7% overall (95% CI 1·8-17·6, P = 0·02), 11·9% in non-doctor, non-nurse HCWs (95% CI 0·9-22·8, P = 0·03). For doctors, using a champion doctor increased uptake by 17·8% (95% CI 7·6-28·0, P < 0·01). No intervention increased uptake by nurses. Increasing uptake requires multi-intervention strategies targeted at different HCW groups.