A disagreement: of course Irene is gay. There's no reason to doubt her. Her behavior (having an affair with the novelist, as well as with his wife, for example) doesn't always match up with her orientation, as she's a paid sex worker. But preference and behavior don't always match up in a lot of people, for a lot of reasons.
Why is the line "I'm gay" included? Because it is an important part of a sub-theme in the episode, which includes Sherlock's discovery that her pulse quickens and her eyes dilate when she is supposedly just pretending to be attracted to him. But it's a theme easy to miss or misunderstand due to cuts in the PBS version. I think that, sadly, deeply-cut PBS versions of Sherlock may prevent Americans from fully appreciating the quality of the writing in this show.
One cut is the end of the Christmas party. There, John, along with Mycroft concerned about the possibility of Sherlock's coming home from the morgue to use drugs, gives up his after-party date to stay home and babysit Sherlock. His girlfriend tells him he's a good boyfriend - to Sherlock. He then babbles something about making it up to her by walking her dog, and she points out that he's got her mixed up with some other woman he's dated and tells him not to call her again. Two other connected cuts are of John at the palace saying he's fighting an urge to steal an ashtray, and then of Sherlock revealing on the way to Irene's that he himself has stolen the ashtray - apparently just to amuse John.
Irene says she's gay, just after John says he's "actually" not. (The "actually" subtly and interestingly affects the meaning. It can suggest the subtext "I may act that way" - for example, your friend caught removing money from your wallet who pleads, "I'm not actually a thief.") Irene then says, "And look at us." She seems to mean that though neither she nor John is normally attracted to men, they both have strong responses, and deep connections, to Sherlock. John pauses to think about that, then audibly smacks his lips. That sound is immediately echoed by the text-alert sound of Irene's smacking her lips and saying, "Ah."
By the way, the exchange between Mycroft and John about not trusting the Secret Service recalls A Study in Pink where John, who has just met Sherlock and has no reason to be loyal to him, refuses to accept money to spy on Sherlock. Mycroft is paying a profound compliment to John's character.
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