Kevin Hart stepped into the spotlight on Sunday night with his usual swagger to accept the Mark Twain Prize for American Humor, occupying a stage lit up with his signature pyrotechnics.

“Can I pee?” Mr. Hart said after a heartfelt tribute from his friend the comedian Dave Chappelle, before waddling offstage at the John F. Kennedy Center for the Performing Arts in Washington. He then reappeared to accept a bust of Mark Twain from David M. Rubenstein, the retiring chairman of the Kennedy Center.

Mr. Hart, 44, is the 25th comic to receive the prize from the Kennedy Center, an honor given annually to the greatest humorists in American comedy. Mr. Hart was joined by his wife and four children, and grinned broadly even as he teared up at bitingly funny roasts and emotional tributes from friends and colleagues in the industry.

“I played arenas with Chris Rock, and I would never play an arena before I saw you do it,” Mr. Chappelle said, crediting Mr. Hart with changing the business of stand-up comedy after a career selling out arena tours and even a football stadium in his hometown, Philadelphia. “You made me dream bigger, and you’re younger than me — it’s humiliating.”

Over a roughly 25-year career — it was noted that he had been doing comedy since the inception of the Mark Twain Prize in the late 1990s — Mr. Hart has sold millions of tickets. He has built a loyal fan base through movies, TV series and many live events — some enhanced by fireworks — including eight comedy specials on relatable narratives, physical comedy and goofy re-enactments. But even when he rags on the cast of characters who file in and out of his life, he is usually the punchline of his own jokes.

His peers also lauded him on Sunday for his work ethic, which includes appearing and casting friends in a slate of Hollywood movies, like the “Jumanji” sequels, dramedies such as “Fatherhood” and “Night School,” and a number of comedic action films.

“I didn’t start out doing what I started to do to get the awards,” Mr. Hart said. “I fell in love with the idea of comedy. It made me say, ‘Oh, my God, I fell in love with something I can do for the rest of my life.’”

The show on Sunday night featured his fellow comics Mr. Chappelle, who was the 2019 recipient of the Mark Twain Prize; Chris Rock, who headlined a tour and subsequent 2023 documentary with Mr. Hart called “Headliners Only”; J.B. Smoove, who met Mr. Hart at the venue where he got his start, the Laff House in Philadelphia; Jimmy Fallon, who performed a tribute in the form of an original country song, complete with heavy twang and a cowboy hat; as well as Jerry Seinfeld, Tiffany Haddish, Regina Hall, Chelsea Handler, Nick Cannon and Keith Robinson. The singer Robin Thicke and rappers Nelly and Dave Burd performed.

The stand-up and sketch group Plastic Cup Boyz, which has opened for Mr. Hart for 17 years, also paid tribute.

Mr. Hart said that receiving the Mark Twain Prize, which is considered by many to be a lifetime achievement award in comedy, meant much more because of the colleagues who showed up to honor him. After poking back at several speakers who took digs at his comedic style, pulled pranks and reminded the audience of his small frame (Ms. Haddish likened the comedian, who is a few inches taller than 5 feet, to a leprechaun), Mr. Hart accepted the award with tears running down his face.

“All jokes aside, thank you for showing up on a night that’s so dope, that could not be as dope without real relationships and real energy,” Mr. Hart said.

The ceremony will be available for streaming on Netflix beginning May 11.