In the course of a public appearance to Lichfield Cathedral on Monday, the King was shouted at by a man who questioned him about Prince Andrew's connections with convicted sex offender Jeffrey Epstein.
The individual inquired how long the King had been known about his brother's association with Jeffrey Epstein, and also raised doubts about whether he had instructed the police to "cover up" for Andrew.
Despite the protest, there were throngs of fans and cheers for the King, who was visiting the region for an event.
Prince Andrew's links with Jeffrey Epstein have been under close examination in recent weeks. The royal, who relinquished his royal duties earlier this month, has categorically denied any misconduct.
"How long have you known about Prince Andrew and Epstein? Have you asked the authorities to cover up for Andrew?"
The monarch remained silent but continued meeting fans, some of whom chanted "Long live the King". One fan can be overheard asking him for a hug.
Another woman outside the venue was also observed telling the heckler to "leave" and to "be quiet".
The scandal over the Duke of York's connections to Jeffrey Epstein has been revived by sexual abuse allegations detailed in Ms. Giuffre's recently released book.
In the publication, the accuser says she had sexual encounters with the prince on multiple times, including once with Jeffrey Epstein and "eight other young girls"
Prince Andrew, who agreed to a financial settlement with the plaintiff in 2022, has consistently denied any misconduct. In 2019, he told BBC Newsnight he did not recall meeting her "at all", and that they "never had any sort of intimate relations."
But in recent times, new inquiries have been raised about how he is able to finance his way of life despite not being a working royal.
The revelation that he has only ever paid a token annual rent on his Windsor mansion has heightened the scrutiny over his living arrangements - even though he made large payments up front, including for renovations.
The Liberal Democrats said he should be called in front of members of parliament to give testimony about his rental agreement of his residence.
But last week, the government declined to give MPs time to discuss the prince's status or his residence, despite increasing calls for investigation.
Buckingham Palace has not issued a statement on whether Prince Andrew might relocate of his current residence - or where he might go.
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