"The answer is no. The vice president had no meetings with individuals associated with the Russian government during the campaign or the transition," Marc Lotter told CNN.
The denial comes after Lotter said Wednesday he was "unaware" of any meetings in an appearance on Fox News, but did not definitively say that none took place.
Speaking to CNN, Lotter explained the comments on Fox News as coming from a place of an overabundance of caution, which made the comments appear unclear.
After speaking generally about Pence's focus on President Donald Trump's agenda, Fox News anchor Bill Hemmer asked Marc Lotter a third time whether any meetings took place.
"Just to nail this down so we're clear, is that a yes or a no? Did he or did he not, and was it relevant in fact?" Hemmer asked.
"I am not aware of anything that I have seen. All of the focus that I saw with Vice President Pence during the campaign has been focused on working the agenda that the people sent him to Washington to accomplish. That's what the vice president is doing," he said.
Lotter also said that the June 9 meeting between Donald Trump Jr. and a Russian lawyer during the campaign that's been under recent scrutiny "happened even before he (Pence) was on the ticket."
Then-Republican nominee Donald Trump selected the then-Indiana governor as his running mate on July 15, 2016.
Pence was not aware of Trump Jr.'s meeting with the Russian lawyer during the campaign, a spokesman for the vice president said Monday. Pence had said in January
that no one from the campaign had any contact with Russians trying to influence the 2016 election.
"The vice president was not aware of the meeting," Lotter told CNN earlier this week
. "He is not focused on stories about the campaign, particularly stories about the time before he joined the ticket."
In the past, Pence has denied any knowledge of contact between the campaign and the Kremlin.
"I joined this campaign in the summer, and I can tell you that all the contact by the Trump campaign and associates was with the American people. We were fully engaged with taking his message to make America great again all across this country. That's why he won in a landslide election," he told Fox News days before the inauguration in January 2017.
Later that morning, in an interview with CBS News, Pence repeated that "of course" there had not been contact between the Trump campaign and Russia, calling allegations of coordination "bizarre rumors."
As CNN previously reported, a number of Pence advisers and staff worked out of Trump Tower during the transition -- and are now worried about being dragged into legal issues. The vice president has hired a lawyer himself.
A senior administration adviser acknowledged last month there have been weeks of hand-wringing behind closed doors among members of Pence's team and an acceptance that the questions about Russia and the Trump campaign were not disappearing. Pence has also been under fire for not knowing a number of things that happened during the transition -- which he led as chairman.