Legal sources said the probe was launched into allegations that local authorities in Lyon had offered illegal help to Macron during his campaign for the presidency last year.
The allegations covered acts of misappropriation of public funds, irregular campaign financing, and abuse of corporate assets.
The complaint cited long-time Lyon mayor Gerard Collomb as one of Macron's earliest political backers who was named interior minister after his election victory in May 2017, Presstv reported.
The complaint alleges that "public resources (human, material and financial) under the control of the city and the Lyon area" were "unduly diverted and put at the disposal" of Macron.
The investigation came following allegations in the French media that Macron had embarked on mounting campaign events at much lower prices thanks to friends and contacts, gaining an unfair advantage over his rivals.
"There is nothing new in this information whose sole purpose is to discredit. The complaint of elected Republicans of Lyon resumed a controversy that the local opposition has been trying to fuel for several months. The city authorities had the opportunity to answer them in detail on several occasions,” an aide to Collomb said last week.
France has strict campaign financing rules that curb spending for the presidential election at $26 million.
Former French president Nicolas Sarkozy has been charged over illegal financing for his successful 2007 campaign which allegedly involved an elaborate system of false billing.