Inflammatory myofibroblastic tumors are uncommon lesions composed of spindled myofibroblasts within a variable background of collagen and inflammatory cells. Although the true nature of these lesions is not fully elucidated, identification of consistent cytogenetic alterations in the anaplastic lymphoma kinase (ALK) gene suggests that they may be neoplastic. A small number of inflammatory myofibroblastic tumors have been reported to harbor human herpesvirus-8 (HHV-8), implicating the virus in its pathogenesis. In this study, 20 cases of pulmonary inflammatory myofibroblastic tumor were analyzed for the presence of HHV-8 with immunohistochemical and molecular methods. In all cases, antibodies to the latent nuclear antigen of the virus were applied. Four open reading frames (ORFs), including ORFs K2, 16, 26, and 72, were targeted utilizing real-time polymerase chain reaction (PCR). The cohort included 9 men and 11 women with a mean age of 37 years (range, 1-81). Microscopically, the tumors were composed of cytologically bland spindle cells with myofibroblastic differentiation. On immunohistochemical studies, 20% of cases (4/20) demonstrated diffuse cytoplasmic positivity with ALK. Immunohistochemical staining for the latent nuclear protein of the virus was negative in all cases (0/20). All tumors (100%, 20/20) tested with real-time PCR were negative for all four ORFs, whereas 100% (10/10) of positive control Kaposi sarcoma cases were positive. Her2 gene expression was present in all (20/20) inflammatory myofibroblastic tumors confirming the presence of amplifiable deoxyribonucleic acid in the tissue lysate. This study documents the absence of HHV-8 in pulmonary inflammatory myofibroblastic tumors, suggesting that further investigation is required to clarify the pathogenesis of this lesion.