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Berberine (BBR) is a plant-derived alkaloid popularly used in a complementary context to treat splenomegaly and chronic disease states such as rheumatoid arthritis, diabetes, and cancer. Recent research has examined BBR as a regulator of lipid and glucose metabolism in association with the AMPK pathway; but little is known concerning its immunological impacts. It is also known that exercise is a potent AMPK activator exhibiting immunological benefits, but nothing is known about the combination of these complementary approaches for treating breast cancer. The purpose of this study is to determine if oral consumption of BBR when paired with physical activity will increase T lymphocyte activation while decreasing the presence of myeloid-derived suppressor cells (MDSC) within the tumor microenvironment, spleen, bone marrow, and blood of the immunocompetent BALB/c 4T1 mammary adenocarcinoma model. We hypothesize that subjects treated with this combination will have reduced MDSC counts, and increased T cell infiltration and activation in tissues of interest, effector, and regulatory subsets. Our data indicates no intervention specific change in NUR77 presence—a transcription factor expressed in antigen specific activation of T cells—in the spleen, lungs and tumor microenvironment of tumor bearing mice. However, T regulatory lymphocyte FOXP3 was also assessed and found to increase significantly in the lungs of tumor-bearing mice. These findings suggest BBR paired with physical activity may have significant immunological implications on T cell activation broadly, and infiltration into the tumor microenvironment. These findings directly inform those who practice complementary and alternative supplementation, an area lacking immunological analysis, including raising concerns for the impact on aberrant, potentially undesirable Treg function.