6:25 PM PT -- A spokeswoman for J.D. Vance tells TMZ ...“J.D. clearly says in the audio that our leaders on the left, especially anti-child radical Randi Weingarten, who brainwash our kids, are deeply disturbing — and he stands by that statement."

Another one of J.D. Vance’s "childless" jibes is coming back to haunt him ... this time slamming teachers who don’t have biological kids ... which fits right in with his controversial "cat lady" remark.

In unearthed audio from 2021, you can hear J.D. ranting about how disoriented and disturbed he feels about leaders of the left without biological kiddos of their own trying to "brainwash the minds of our children."

Listen to the clip -- J.D. specifically targets American Federation of Teachers President Randi Weingarten ... saying "she doesn't have a single child" ... and urges her to have children of her own and "leave ours the hell alone."

Important to note, Weingarten does have two adopted kids ... but Vance has a history of not seeing adopted or stepchildren as "real" kids.

It was only last month when J.D.’s "cat ladies" comment resurfaced when he took a swipe at Democratic leaders ... saying the country was being run by people like VP Kamala Harris and Rep. Alexandria Ocasio-Cortez, who haven’t had kids.

He later said he didn't regret the remark ... adding people were totally missing his point, calling it all just flat-out sarcasm.