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Catherine may have been involved during her marriage to the King with Henry’s favourite male courtier, Thomas Culpeper, a young man who “had succeeded [him] in the Queen’s affections”, according to Dereham’s later testimony. She had considered marrying Culpeper during her time as a maid-of-honour to Anne of Cleves.

Culpeper called Catherine “my little, sweet fool” in a love letter. It has been alleged that in Spring 1541 the pair were meeting secretly. Their meetings were allegedly arranged by one of Catherine’s older ladies-in-waiting, Jane Boleyn, Viscountess Rochford (Lady Rochford), the widow of Catherine’s executed cousin, George Boleyn, Anne Boleyn’s brother.

People who claimed to have witnessed her earlier sexual behaviour while she lived at Lambeth reportedly contacted her for favours in return for their silence, and some of these blackmailers may have been appointed to her royal household.

John Lassels, a supporter of Cromwell, approached the Archbishop of Canterbury, Thomas Cranmer, telling him that his sister Mary refused to become a part of Queen Catherine’s household, stating that she had witnessed the “light” ways of Queen Catherine while they were living together at Lambeth. Cranmer then interrogated Mary Lassels, who alleged that Catherine had had sexual relations while under the Duchess of Norfolk’s care, before her relationship with the King.

Cranmer immediately took up the case to topple his rivals, the Roman Catholic Norfolk family. Lady Rochford was interrogated and as she feared that she would be tortured, she agreed to talk. She told how she had watched for Catherine backstairs as Culpeper had made his escapes from the Queen’s room.

During the investigation a love letter written in the Queen’s distinctive handwriting was found in Culpeper’s chambers. This is the only letter of hers that has survived (other than her later “confession”).

On All Saints’ Day, November 1, 1541, the King arranged to be found praying in the Chapel Royal. There he received a letter describing the allegations against Catherine. On November 7, 1541 Archbishop Cranmer led a delegation of councillors to Winchester Palace in Southwark, to question her.

Even the staunch Cranmer found the teenaged Catherine’s frantic, incoherent state pitiable, saying, “I found her in such lamentation and heaviness as I never saw no creature, so that it would have pitied any man’s heart to have looked upon her.” He ordered the guards to remove any objects she might use to commit suicide.

Catherine was stripped of her title as Queen on November 23, 1541 and imprisoned in the new Syon Abbey, Middlesex, formerly a convent, where she remained throughout the winter of 1541. She was obliged by a Privy Councillor to return the ring previously owned by Anne of Cleves, which the King had given her; it was a symbol of removal of her regal and lawful rights. The King would be at Hampton Court, but she would not see him again. Despite these actions, her marriage to Henry VIII was never formally annulled.