While Gleeson stole the headlines, it was India who inevitably won the match by 49-runs. The victory gives them an unassailable 2-0 lead heading into the third and final match at Trent Bridge.
The hosts subsided to 121 all out in just 17 overs as they produced yet another limp batting display.
The tourists kicked off proceedings by setting England a target of 170. Chris Jordan, who took four for 27, and Gleeson both enjoyed a particularly good day in the field.
The 34-year-old became the oldest player to make his England debut since Paul Nixon in 2007. He marked the occasion by taking the wickets of Rohit Sharma, Virat Kohli and Rishabh Pant within the space of four deliveries.
Gleeson's career almost came to an end in 2020 following a stress fracture in his lower back.
The player signed a T20-only deal with Lancashire this season and has proved a revelation in the Vitality Blast.
"It wasn't me contemplating retirement," he said. "It's something that would have been forced upon me.
"(The recovery) was a long, slow process. Unfortunately because of my age it takes a little bit longer to heal from things like that.
"But (playing again) makes it all worthwhile. Just getting back out in the park and playing for Lancs was what my aim was and to do that was great. And then to do well was even better.
"But this wasn't even anywhere near my radar. So to get the nod first of all, and then to actually go and perform as well, even better.
"It's all about winning games of cricket at the end of the day so it's disappointing to lose but on a personal note it was a great start."
Renowned for his consistency and being able to nail his yorkers in pressure scenarios, Gleeson showed injury has not taken the edge off as he approached 90mph on the speed gun on Saturday.
"You've just got to stick with what you do," he said. "There's no point in coming in and trying something completely different.
"That's what I've been picked for. That's what I've been told to go out and do so to execute is always great."