Keys and her Keep A Child Alive charity will honour Winfrey's work with humanitarian causes on November 1 at New York's Hammerstein Ballroom.
'Love on Top' singer Beyoncé will play an intimate set during the gala and also offer her own thoughts on how Winfrey has positively influenced US culture.
Carole King, Angélique Kidjo, Alabama Shakes and Keys herself are also scheduled to perform at the Black Ball.
Keys said of the benefit concert's lineup: "Women will be rocking the house this year!"
The Keep a Child Alive charity has previously honoured the late Beatle George Harrison and former US President Bill Clinton.
Listen Alicia Keys's song 'Girl on Fire' below: