Hairy cell leukemia (HCL) exhibits a characteristic immunophenotypic profile that is strongly positive for pan-B-cell markers; positive for CD103, CD11c. and CD25; and usually negative for CD5, CD10, and CD23. We evaluated 35 HCL cases and identified atypical immunophenotypes in 12 cases (34%), including CD103- in 2 (6%), CD25- in 1 (3%), CD10+ in 5 (14%), and CD23+ in 6 (17%) cases. Among these cases one was CD103-/CD10+ and one was CD10+/CD23+. All available specimens from the 12 cases were reviewed and showed morphologic features characteristic for HCL. The initial clinical information was reviewed and showed no significant differences with that reported for typical HCL. Of the 12 cases, 11 patients received purine analogue therapy and achieved complete remissions. Our study indicates that it is not uncommon for HCL to display an unusual immunophenotype, including negativity for CD103 or CD25. Recognizing the variability of immunophenotype and correlating with morphologic and clinical features are essential for establishing an accurate diagnosis of HCL.