Physical function limitations have been associated with poor health outcomes, which have a negative impact on quality of life of older individuals. This study examined the association between depression, viral load, and acculturation with physical function among Latino men living with HIV. A secondary data analysis was performed using a cross-sectional data of 146 Latino immigrant men living with HIV in New York City and Washington, DC. Physical function was measured using the Short-Form Health Survey (SF-12). Uncontrolled HIV infection and depression were associated with worse physical function, thus implying the importance of adequate health care to address these conditions. Preserving physical function should start during middle adulthood, particularly among people living with HIV because of their greater risk of developing age-related challenges such as depression, diabetes, cardiovascular diseases among others. This study informs future interventions to preserve physical function and achieve the goal of successful aging.