As mentioned earlier in the responses, lugging to low while cruising will drop you below the efficiency range of the engine and eliminate an furl savings. BUT... since the stock engine makes peak torque at 1800 rpm, you'd be hard pressed to be keeping your rpm below that on the highway. Unless maybe you were just taking a nap on the shoulder?
The biggest fuel mileage gain I have personally seen was years ago with a customer years ago who drove like an old lady and was meticulous about logging his fuel fills/mileage and everything else. After "power build" which included a compression bump, headers, intake and small 4bbl carb AND most importantly a longer duration/higher lift camshaft, he saw his highway fuel consumption for his (stock geared on stock tires FJ60) drop from 13mpg to just over 16mpg. A 25% gain!
I attribute that primarily to the change in camshaft. The factory camshaft is well beyond it's efficiency range when cruising at North American highway speeds. The camshaft bumped the efficiency range upward into a more appropriate area for how the rigs gets used here. Mr Toyoda and friends expected the old F engines to spend their lives creeping around backroads, fields and village streets in the third world, not running down multilane highways in the US. To the best of my knowledge they never changed the camshaft specs over the years. if they did it was still not enough.
To your original question... around town You will almost certainly not see any improvement. For pure highway runs if you drive gentle, just relaxing in the right hand lane, I would hopefully expect to see about 10% or so gains in the long term. In flat country. In the hills and mountains, all bets are off. If you push the rig in terms of speed/acceleration/"sportiness", you will never see any difference with the 5 speed.
Mark...