The integration of nanofibers into conventional fabrics may open up new opportunities such as improving the comfort performance and thermal management properties of outdoor clothing. Nanofibers are able to form a highly porous mesh and their large surface-to-volume ratio improves performance for many applications. This study shows the possible utility of the nanofiber coating on conventional knitted fabrics for improving the wind-resistance and breathability properties. It was seen that nanofiber coating did not cause a significant effect on water vapor and thermal resistance of electrospun thermoplastic polyurethane nanofiber coated cotton (CO), modal (CMD), viscose (CV), and lyocell (CLY) single jersey fabrics, while resistance to air permeability was increased with the increased nanofiber coating. High level of air resistance was achieved with 30 min of coating. In terms of comfort properties, the nanofiber coating proved to be advantageous due to its lower air permeability with its water vapor permeable structure. However, thermal insulation level of these fabrics was still low and fragile nanofiber layer needed to be protected. Therefore, a multi-layered fabric form was derived from combination of cotton and lyocell fabrics with a nanofiber layer. The results showed that nanofibers could be used to improve the wind-resistance and comfort properties of multi-layered knitted structures.