Crosslinguistic studies on the comprehension and production of wh-questions in spoken languages report an advantage for subject over object clauses. To date, very few studies have been carried out to investigate the presence and direction of the alleged subject/object asymmetry of wh-questions in sign languages. The present study analyses the comprehension of Italian Sign Language (LIS) wh-questions, featuring the wh-sign ‘which’, through a sentence-to-picture matching task administered to three groups of participants: hearing native signers (so-called CODAs), deaf native signers, and deaf non-native signers. The task investigates two types of structures: potentially ambiguous which-questions displaying the stranding of the nominal restrictor, and which-questions featuring a disambiguating strategy. Results show a preference for an object interpretation both in ambiguous and unambiguous which-questions driven by animacy features.