The 16th Infantry Division was formed for the first time in late 1915 or early 1916, following the Greek mobilization on 10 September 1915, in response to the mobilization of Bulgaria.
At the commencement of the Greco-Italian War in October 1940, it was reactivated in Lamia as a brigade but was quickly expanded to a division-strength by the end of the year and placed under the command of the Western Macedonia Army Section (III Army Corps).