- Viola – castaway, disguised as a eunuch called Cesario, in service to Orsino. A young woman of aristocratic birth, and the play's protagonist. Washed up on the shore of Illyria when her ship is wrecked in a storm, Viola decides to make her own way in the world. She disguises herself as a young man, calling herself "Cesario," and becomes a page to Duke Orsino. She ends up falling in love with Orsino — even as Olivia, the woman Orsino is courting, falls in love with Cesario. Thus, Viola finds that her clever disguise has entrapped her: she cannot tell Orsino that she loves him, and she cannot tell Olivia why she, as Cesario, cannot love her. Viola's poignant plight is the central conflict in the play. She is also fluent in French.
- Orsino – Duke of Illyria, wooing Olivia. A powerful nobleman in the country of Illyria. Orsino is lovesick for the beautiful Lady Olivia, but finds himself becoming more and more fond of his handsome new page boy, Cesario, who is actually a woman — Viola. A supreme egoist, Orsino mopes around complaining how heartsick he is over Olivia, when it is clear that he is chiefly in love with the idea of being in love and enjoys making a spectacle of himself.
- Olivia – a countess, resisting Orsino's wooing. A wealthy, powerful, and noble Illyrian lady. Olivia is courted by Orsino and Sir Andrew Aguecheek, but to each of them she insists that she is in mourning for her recently deceased brother and will not marry for seven years. Olivia and Orsino are similar characters in that each seems to enjoy wallowing in his or her misery. Viola's arrival in the masculine guise of Cesario enables Olivia to break free of her self-indulgent melancholy.
- Sebastian – Viola's lost twin brother. When Sebastian arrives in Illyria, travelling with Antonio, his close friend and protector, he discovers that many people seem to think that they know him. Furthermore, the beautiful Lady Olivia, whom Sebastian has never met, wants to marry him.
- Malvolio – the straitlaced steward — or head servant — in the household of Lady Olivia. Malvolio is very efficient but also very self-righteous, and he has a poor opinion of drinking, singing, and fun. His priggishness and haughty attitude earn him the enmity of Sir Toby, Sir Andrew, and Maria, who play a cruel trick on him, making him believe that Olivia is in love with him. In his fantasies about marrying his mistress, Malvolio reveals a powerful ambition to rise above his social class.
- Maria – Olivia's clever, daring young serving-woman. Maria is remarkably similar to her antagonist, Malvolio, who harbors aspirations of rising in the world through marriage. However, Maria succeeds where Malvolio fails — perhaps because she is more in tune than Malvolio with the anarchic, topsy-turvy spirit that animates the play.
- Sir Toby Belch – Olivia's drunken uncle. Olivia lets Sir Toby live with her but does not approve of his rowdy behavior, practical jokes, heavy drinking, late-night carousing, or friends (specifically the idiotic Sir Andrew). But Sir Toby has an ally — and eventually a mate — in Olivia's sharp-witted serving-woman, Maria. Together, they bring about the triumph of fun and disorder, which Sir Toby embodies, and the humiliation of the controlling, self-righteous Malvolio.
- Sir Andrew Aguecheek – a simple-minded companion of Sir Toby. Sir Andrew Aguecheek attempts to court Olivia, but he has no chance of success. He thinks that he is witty, brave, young, and good at languages and dancing, but he is thought to actually be a complete idiot.
- Feste – the clown, or court jester, of Olivia's household. Feste, also known as The Fool, moves between Olivia's and Orsino's homes, earning his living by making pointed jokes, singing old songs, being generally witty, and offering good advice cloaked under a layer of foolishness. In spite of being a professional fool, Feste often seems the wisest character in the play.
- Valentine – gentleman attending on Duke Orsino.
- Curio – gentleman attending on Duke Orsino.
- Fabian – a servant and friend to Sir Toby. He assists Maria and Sir Toby in their plot to humiliate Malvolio.
- Antonio, a captain and friend to Sebastian. He rescues Sebastian after his shipwreck. Antonio has become very fond of Sebastian, caring for him, accompanying him to Illyria, and furnishing him with money - all because of a love so strong that it seems to be romantic in nature. When the principal characters marry at the end of the play, Antonio is left out, his love for Sebastian unrequited.
- Captain, friend to Viola, rescues Viola after the shipwreck. He helps Viola become a page to Duke Orsino and keeps her identity a secret.
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