Disseminated Acanthamoebiasis poses a grave threat that can potentially leads to fatal outcomes in immunosuppressed individuals. This study investigates the immunological and histopathological effects in immunosuppressed animals after a systemic infection by A. polyphaga strain isolated from contaminated contact lenses. Fifty male Swiss albino mice were divided into three groups: immunosuppressed control (CS), immunosuppressed A. polyphaga infected (AS), and negative controls (C). The CS and AS groups were sacrificed at 2, 7, and 15 days post-infection. The results showed that the AS mice displayed a range of clinical signs including emaciation, changes in coat color, ulcerations, enlargement of the testicular region, circular moving, bilateral eye lesions, diminished mobility, and alterations in posture. There was a highly significant increase in the cytokines IL-1β, TNF-α, and IL-10 levels, especially at the onset of infection in AS groups. The infected liver tissues demonstrated an inflammatory response, characterized by an intercellular distribution of parasite stages, multiple focal areas of inflammatory cell infiltration, diffuse vacuolar degeneration with sporadic hepatocyte necrosis, dilatation of blood sinusoids, and congestion in the blood vessels. Additionally, significant changes were observed in liver enzyme levels. These findings indicated the potential impact of systemic infection by A. polyphaga in immunosuppressed hosts, highlighting the need for awareness and intervention strategies to mitigate such life-threatening infections. Further research is crucial to better understand the disseminated Acanthamoebiasis by analyzing changes in gene and protein expressions in liver tissues.