Jacksonville is about two-and-a-half hours by car from Kissimmee.
The Kissimmee Police Department said via its official Twitter site Friday night that the officers were shot in the area of Palmway and Cypress. One officer remains in grave condition, according to Kissimmee police.
Four suspects were involved in the shooting that took place in an area known for drug activity. One suspect remained at large late Friday.
Earlier, Florida Gov. Rick Scott had tweeted condolences amid initial reports that two officers were killed in Kissimmee.
Scott then tweeted about the officers shot in Jacksonville. "Just learned of two @JSOPIO in danger tonight - we stand with ALL law enforcement in Florida."
According to the Jacksonville Sheriff's Office, one of the officers shot in that city was in critical condition and the other was in stable condition. The suspect was shot and killed by police, authorities said.
No further information was immediately available.
The Associated Press contributed to this story.