Atlantic salmon (Salmo salar) is repeatedly exposed to and infected with ectoparasitic salmon lice (Lepeophtheirus salmonis) both in farms and in nature. However, this is not reflected in laboratory experiments where fish typically are infected only once. To investigate if a previous lice infection affects host response to subsequent infections, fish received 4 different experimental treatments; including 2 groups of fish that had previously been infected either with adult or infective salmon lice larvae (copepodids). Thereafter, fish in all treatment groups were infected with either a double or a single dose of copepodids originating from the same cohort. Fish were sampled when lice had developed into the chalimus, the pre-adult and the adult stage, respectively. Both the specific growth rate and cortisol levels (i.e. a proxy for stress) of the fish differed between treatments. Lice success (i.e. ability to infect and survive on the host) was higher in naïve than in previously infected fish (pre-adult stage). The expression of immune and wound healing transcripts in the skin also differed between treatments, and most noticeable was a higher upregulation early in the infection in the group previously infected with copepodids. However, later in the infection, the least upregulation was observed in this group, suggesting that previous exposure to salmon lice affects the response of Atlantic salmon towards subsequent lice infections.