Canadian international wing Winston Stanley helped himself to a quick-fire brace of tries in the first half for the Yorkshiremen in the continual absence of Dan Scarbrough.
Two penalties and a conversion from Braam van Straaten took the home tally to 18 points in the opening half, while Gloucester had just two penalties from England Sevens captain Simon Amor to their credit.
In the second half, back rower Alix Popham and Scottish fly-half Gordon Ross added tries for the Tykes, Van Straaten converting Ross's effort.
A third penalty from Amor and then a late try from wing Tom Beim got Gloucester on the second-half scoreboard, but even with the conversion from Henry Paul they were still 15 points behind the eventual winners, having been without sin-binned centre Terry Fanolua for 10 minutes.
They did claw back a bonus point with a late converted try from wing Darren O'Leary, but the win was not to be for the visitors despite their late rally.
There will not be too many tears shed at Kingsholm however, having already scooped the regular season top spot, with only the Zurich Championship Final at Twickenham standing between them and the title.
Leeds meanwhile still harbour a mathematical chance of a play-off position, lying four points behind third-placed Sharks, having played the same number of games.
For Leeds Tykes:Tries: Stanley 2, Popham, RossCons: Van Straaten 2Pens: Van Straaten 2
For Gloucester:Tries: Beim, O'LearyCons: Paul 2Pens: Amor 3