Yes.

The were were multiple cases of US pilots flying various marks of the Supermarine fighter. There were the three so-called Eagle Squadrons - No. 71, No. 121, No. 133- which were composed of US volunteer pilots and formed over the course of 1940-41. The three squadrons flew Spitfires and were enfolded into the USAAF sometime after Pearl Harbor. The 31st Fighter Group of the USAAF also used Spitfires in the North African and Mediterranean theaters when their P-39s were found unsuitable for combat conditions. The US's 68th and 7th Photo Groups also flew the PR.XI unarmed reconnaissance version of the Spitfire within Europe.

So the Spitfire was used by several US units during the war. In most cases, the Spitfire was a substitute when there were no American-built planes of equivalent performance available.

Or bf-109’s