Harrison, 39, wrote the following in an Instagram post that featured photos of his children:
"I've missed way too much for way too long...and I'm done. Many thanks to my family, coaches, the fans, and everyone who played a role in my football life."
Harrison, who will turn 40 on May 4, previously announced his retirement in August 2014 before returning to the Pittsburgh Steelers the following month.
A 2008 NFL Defensive Player of the Year and five-time Pro Bowl selection (2007-11), Harrison is perhaps best remembered for his 100-yard interception return for a touchdown in Super Bowl XLIII against the Arizona Cardinals.
Harrison collected a franchise-record 80.5 sacks in 176 career games with the Pittsburgh Steelers, who released him in December after playing just 40 snaps over 14 games.
He signed with the New England Patriots on Dec. 26 and started for the team in its 41-33 loss to the Philadelphia Eagles in Super Bowl LII.
Harrison was undrafted out of Kent State in 2002 and played for NFL Europe's Rhein Fire the following year. He spent his first 10 NFL seasons with the Steelers before joining the rival Cincinnati Bengals in 2013.
Harrison collected 793 tackles, 84.5 sacks, 34 forced fumbles, eight fumble recoveries and eight interceptions in 192 career regular-season games.