Background: Boeravinone B (BB), a therapeutic biomarker isolated from roots of Boerhavia diffusa, possesses immunomodulatory properties. Objective: In the present study, we investigated cytotoxicity and immunomodulatory effect of BB on human dendritic cells (DCs). Materials and Methods: In this study, buffy coat of human healthy volunteers was collected and the monocytes were separated. Effect of various concentrations of BB on DCs from induced differentiation of monocytes was studied on day 7 using ELISA microplate reader. The effect of BB on the immature DCs generated in the presence and absence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin4 (IL4) was also noted. The binding of BB with the DC maturation markers CD80, CD83, and CD86 including antigen uptake were evaluated using fluorescent-activated cell sorter (FACS). Results: Viability of DCs was not affected at BB concentration of 100 μg/ml. Phytohemagglutinin (PHA), BB showed increase in DC expression of CD80 expression up to 6.7% (P < 0.001) and 7.27% (P < 0.001) and increase in antigen uptake up to 7.67% (P < 0.001) and 8.87% (P < 0.001) when compared with dimethyl sulfoxide control. PHA and Boeravinone B supplemented with GM-CSF and IL4 showed increase of CD83 expression up to 6.86% (P < 0.01) and 4.63% (P < 0.05). However, PHA and BB showed increase of CD86 expression up to 6.6% (P < 0.01) and 6.73% (P < 0.01) when compared with control. Conclusion: The results justify that BB induced differentiation of monocytes into mature DCs.