A German non-government group targeted by the US measures, HateAid, labelled them an "act of repression".
Washington announced sanctions on Tuesday against Thierry Breton, the former top tech regulator at the European Commission, and four others -- including two members of the German group.
It accused them of promoting "censorship crackdowns by foreign states -- in each case targeting American speakers and American companies".
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Breton was described by the State Department as the "mastermind" of the EU's Digital Services Act (DSA) that imposes content moderation and other standards on major social media platforms operating in Europe.
Also subject to the ban were Anna-Lena von Hodenberg and Josephine Ballon of HateAid, a German organisation the State Department said functions as a trusted flagger for enforcing the DSA.
"The entry bans imposed by the USA, including those against the chairpersons of HateAid, are not acceptable," German Foreign Minister Johann Wadephul wrote in a post on X.
"The DSA was democratically adopted by the EU for the EU – it does not have extraterritorial effect," he added.
HateAid in a statement called the US government decision an "act of repression by an administration that increasingly disregards the rule of law and tries to silence its critics with all its might"
"We will not be intimidated by a government that uses accusations of censorship to silence those who stand up for human rights and freedom of expression," it added.
Washington has scaled up its attacks on EU regulations after Brussels earlier this month fined Elon Musk's X for violating DSA rules on transparency in advertising and its methods for ensuring users were verified and actual people.
The US visa ban also targeted Imran Ahmed of the Center for Countering Digital Hate, a nonprofit that fights online hate, and Clare Melford, who leads the UK-based Global Disinformation Index.
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