Supporters of LGBT, Muslim, and Latino rights gathered outside Trump Towers in New York to protest the presumptive Republican nominee.
The group also criticized the former reality TV star's "use of the recent massacre at a gay bar in Orlando to polarize Americans."
Advocates organized a "die in," where protesters laid on the ground and simulated corpses, to speak out against recent acts anti-gay acts of violence.
CNN's Jim Acosta said it was the largest protest of its kind that he'd seen.
Trump responded to the massacre of 49 people at Pulse nightclub by attacking Muslims and suggesting that the U.S. engage in racial profiling.
"I hate the concept of profiling, but we have to start using common sense," he said on Sunday.
Orlando shooter Omar Mateen was Muslim and had pledged his allegiance to the so-called Islamic State. However, as House Speaker Paul Ryan said,"there’s a really important distinction ... this is a war with radical Islam — it’s not a war with Islam. Muslims are our partners."
Protesters at Tuesday's rally demanded an end to Trump's hateful rhetoric.
“The LGBTQ community is a rainbow coalition of races and religious beliefs. While we stand united against any kind of religious bigotry, we will not stand silent while our Muslim LGBTQ brothers and sisters continue to be targets of hate speech,” ACTUP said in a statement.
Trump, who has referred to himself as a supporter of LGBT rights, met with evangelical leaders on Tuesday. One of those was Tony Perkins, the president of the Family Research Council, a group that opposes gay rights and whose leaders often make derogatory comments about LGBT people.
“By continuing to embrace, rather than condemn these hate groups, Donald Trump and the Republican Party are spitting on the victims of the Orlando Massacre," the group said.