Exxon Mobil Corp. has sued two Cuban companies in a U.S. federal court under a newly revived provision that permits legal action by U.S. citizens or entities against companies doing business on property that was confiscated by the Cuban government.

The lawsuit, filed Thursday in the U.S. District Court for the District of Columbia, claims that two Cuban companies, Corporación Cimex S.A. and Unión Cuba-Petróleo, use and profit from property, including oil refineries and service stations, that an Exxon Mobil predecessor company owned before it was seized in 1960 by the Fidel Castro government.

Exxon Mobil is hoping to recover about $280 million in losses associated with the property, a company spokesman said.

Cimex didn’t immediately respond to a request for comment Friday. A Unión Cuba-Petróleo representative said Friday she can’t comment without authorization.

Havana-based Cimex operates in industries including real estate, financial services and logistics, according to its website.

Unión Cuba-Petróleo, or Cupet, is an oil company and operates refineries in Cuba, according to its website. Exxon Mobil said Cimex operates hundreds of service stations in cooperation with Cupet, according to the complaint.

The Exxon Mobil case is one of the first lawsuits filed under Title III of the 1996 Helms-Burton Act since its two-decade suspension was lifted. Two U.S. citizens filed suit Thursday against Carnival Corp. , alleging the cruise operator is doing business on seized property in Cuba that they had stakes in.

Title III allows certain U.S. nationals and companies with claims to properties confiscated by the Cuban government to sue companies, including some Cuban entities, that are operating on that property for compensation. The law applies to properties confiscated in or after 1959.

The provision had been suspended in response to complaints by the European Union that the law overreached its jurisdiction and restricted the EU’s trade with Cuba.

The Trump administration first in March partially lifted the suspension to allow private actions against Cuban entities or subsidiaries that are identified on the State Department’s restricted entities list, before fully lifting the suspension on Title III beginning Thursday.

Cimex is on the State Department’s restricted list and is also blacklisted by the U.S., which said in 2004 that it was owned by the Cuban government and no U.S. companies or individuals are permitted to work with the company.

Exxon Mobil essentially inherited the claim of ownership on the Cuban property from Standard Oil Co., according to the complaint. Standard Oil is a predecessor to Exxon Mobil.

The U.S. Justice Department’s Foreign Claims Settlement Commission certified that Standard Oil suffered a loss of more than $71.6 million as a result of the confiscation, which is one of the highest among the nearly 6,000 certified claims in the agency’s Cuban claims program.

Exxon Mobil is seeking damages that include the value of the claim, plus interest, among other things, according to the lawsuit.

Write to Mengqi Sun at mengqi.sun@wsj.com