Good job for checking your vehicle and trailer numbers for real rather than assuming things are OK!
Since you mention the possible removal of a bike, I'm guessing the weights were done at different times? I didn't see the confirmed re-weighing of the highlander alone, and it looks like a difference of 10 gallons of gas in the highlander's gas tank could change its weight by over 60 lbs, changing part of your equation.
Another alternative: I don't know how your hitch works, but on ours we have an approximate amount of tightening of the WDH so although it's close to the same every time, it's never identical.
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2003 3124KS
240 W solar (thx to previous owner)
Platinum cat (thx to previous owner)
Tacoma 4-door long-bed
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