- Viola, twin sister to Sebastian. When disguised as a man, she is known as Cesario
- Orsino, Duke of Illyria, in love with Olivia
- Olivia, a Countess
- Sebastian, twin brother to Viola
- Maria, a gentlewoman in Olivia's household
- Sir Toby Belch, Olivia's uncle
- Sir Andrew Aguecheek, a companion of Sir Toby's
- Malvolio, steward to Olivia
- Feste, also referred to as the Fool, a clown
| - Fabian, a member of Olivia's household.
- Antonio, a captain, a friend to Sebastian.
- Captain, a sea captain who helps Viola.
- First Officer, an officer sent from Duke Orsino to arrest Antonio.
- Second Officer, an officer who helps arrest Antonio.
- Valentine and Curio, two gentlemen attending Orsino
- Priest, a Holy Father
- Servant, a servant who reports that Viola/Cesario has returned to see Olivia
- Musicians, Lords, Sailors, and other attendants
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