The accounts contain 16 million Euros and no access will be allowed to the money whilst Vatican authorities investigate the sale of Vatican-owned real estate.
Accusations against the three relate to property deals between 2001 and 2008 when 28 Vatican buildings were sold to Italians.
It's the latest in a list of investigations launched at the Vatican Bank into alleged corruption.
Since the election of Pope Francis the bank has been ordered to be more transparent.
Last month the Vatican gave staff a manual on economic ethics and accountability.