There is an abundance of Ho Yay in Ultimate Spider-Man:
- This line:Spider-Man: I hate you, Deadpool.
Deadpool: I know you mean "love"!
- "Ultimate Deadpool" may have been one episode, but there have been Shout Outs to the Spideypool fandom. The line above, their interactions seeming like a love/hate relationship, and the fact that Deadpool is constantly in Spidey's personal space (even pulls his face in close enough to kiss him).
- A little between Blade and Werewolf by Night.
- Peter and Harry all the way. Epecially since MJ was a.) barely used as a character for the first several seasons and b.) has showed no romantic inclination towards Peter. Spidey is constantly thinking about Harry throughout the first two seasons. Spidey also tends to gush about Harry fairly often in the first season. He also tends to put Harry above everything else and has undying faith and trust in him, to the point that Harry is the only person outside the original team (who already knew and without his permission) that he even remotely considers revealing that Spider-Man and Peter Parker are two sides of the same coin to, whereas he'll act somewhat horrified over the prospect of others finding out. And yes, the horrified reaction includes both Aunt May, the closest thing he has to a mother, and MJ, who he's know for significantly longer than Harry. On Harry's side, he's prone to fits of jealously towards other people that Peter is focusing on instead of him, always forgives Peter for any transgressions (perceived or otherwise,) and trusted Peter enough to reveal that he was Venom. This isn't even getting into the fact that they both seem to have a mutual dependency on each other.
- When Harry is about to be taken by Carnage's hive, it's the revelation that Peter is Spider-Man that allows him to take control. When it's revealed Anti-Venom needs a host, Peter tries to sacrifice himself to save Harry, but Harry takes the Symbiote knowing it has to be him. Peter is distraught hearing this. When the hive is destroyed, Peter dives in and saves him, shouting his name and begging him to wake up. When Harry does, Peter is relieved. Note the fact that Peter is carrying him bridal style and touching his cheek as he begs for him to wake up. Plus, unlike the other versions of him, Harry actually is happy that Peter is Spider-Man.
- Peter and Flash start to get a one sided case of this after the latter begins his character development. Flash already started off being a Spidey fanboy, and after he stops bullying Peter they're shown getting rather chummy from time to time. After Flash becomes Agent Venom it arguably gets worse as we get to see him fanboying even and more often, and get to witness his sheer excitement over being on the team. Then after he finds out Peter and Spider-Man are the same during the season 3 finale he's shown to be shocked, but then portrayed as being even closer in latter episodes. Comes to a head in Anti-Venom where he acts like a jealous boyfriend due to Spidey gushing about Harry (of course) and unintentionally throwing it in Flash's face. A notable moment includes Flash pointing out to Harry that Peter called him a "good friend" as he's being violently assaulted by the Anti-Venom.
- A lot of the resentment Flash holds against Scarlet Spider seems to be more to do with the later's betrayal of Peter than anything else, emphasizing how much Flash care about him.
- There's a certain degree of Ho Yay between Iron Fist and Spidey as well. Out of his original teammates Peter bonds with quickly and gets along best with Danny, and they're shown to value each other highly. Enough so on Spider-Man's end that he once followed Danny across the world to convince him not to leave the team.
- The rivalry between Nova and Spider-Man in the first season has potential to come off as Unresolved Sexual Tension depending on the viewer. Also as Nova gets nicer in the second season he can come off as being somewhat Tsundere towards Spidey.
- Spider-Man once referred to Nova as "My little light bulb." Yes, really.
- Spider-Man's reaction to meeting Iron Man. That is all.
- While it was clearly meant to be platonic the fact that Hulk can be characterized with the Tsundere trope in regards to Spidey arguable qualifies as this.
- Spider-Man seems oddly drawn to older male figures during the first two seasons. While this is a likely result of losing Uncle Ben (and therefore he may have subconsciously been looking for a male role model) Peter can get oddly touchy-feely with some of these characters. To the point of having No Sense of Personal Space in a couple of instances.
- Honestly Spider-Man has this going with most of his male teammates (and a few of his villains) on varying levels throughout the show's run.