When the Persians seized the state of Lydia in 456 B.C., Lesvos came under their control. In 527 it took part in the campaign of Kamvysis, King of the Medians and the Persians, against Egypt and in 513 it assisted Darius against the Scythians. In 499 it joined in the revolution of the Ionians against the Persa and in 492 was subjected by the Persians. In 479 after the battle of Mykalis, it threw off the Persian yoke and participated in the Attic-Eleian Alliance.
During the Peleponnesian War, with the exception of Mythimna, the island revolted against the Athenians, as it took the side of Sparta and in 427 the Athenian general Pachis the Epicure conquered the island. In 405 Lesvos succumbed to the Spartans who were led by Lysander, but it was retaken by the Athenians in 389. In 385 it came again under Spartan dominance, but in 375 Lesvos joined the second Athenian alliance. After the battle of Granikos, the inhabitants of Mitilini allied themselves with Alexander the Great. The brothers, Erigyios and Laomedon, from Lesvos were childhood friends of Alexander and he bestowed high ranks on them. After the death of Alexander in 323 B.C., Lesvos came under the Ptolemies of Egypt and in 88 BC the Romans conquered the island.