This study investigated the psychometric properties of the Humor Styles Questionnaire (HSQ) and the relation between humor and psychological well-being within the context of Italy. A total of 293 (178 females, 115 males) Italian high school and undergraduate university students – whose ages ranged from 14 to 25 years – completed the Italian versions of the HSQ and the Ryff’s Psychological Well-Being scales (RPWB). The HSQ scale reliabilities were generally acceptable, and intercorrelations among the scales were rather low; the confirmatory factor analysis supported the four-factor structure. Males reported significantly more use of Aggressive humor than did females; no differences were found between adolescents and young adults in the use of humor styles. Affiliative and Self-enhancing humor styles were positively associated with the six dimensions of the RPWB, whereas Self-defeating humor was negatively correlated with the RPWB scales. SEM analysis showed a significant and positive relationship between humor as measured by the HSQ and psychological well-being as assessed by the RPWB. Overall, the findings supported the theoretical structure and usefulness of the HSQ in an Italian context and the differential role of humor components in the various dimensions of psychological well-being.