Obviously, legally, they can. What I'm wondering is can they politically plead the fifth? And I'm thinking specifically about Ambassador Sondland, who's been placed at the middle of all this and in direct communication with Trump by multiple people. Trump said in 2016, "If you're innocent, why are you taking the Fifth Amendment?". This was during the Clinton investigation, and for once he had a point. When people around you take the fifth, it looks shady as hell. Clinton's IT staffer did it, and it looked really bad. I don't think Trump can survive politically if someone like Sondland pleads the fifth. That will cement in everyone's mind the certainty that illegal stuff was going on. Would any Trump supporters here be OK with someone like Sondland or Bolton pleading the fifth about this Ukraine business?
When their are direct witnesses to the question you maybe taking the fifth to it would be pointless.Sondland has to be very careful, he has already had one do over on his testimony, a second might lead to a perjury charge.