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The objective of the study was to evaluate the immunomodulatory effect of isolate compound Kaempferol-5-O-β-D-glucopyranoside from methanol extract of aerial parts of Indigofera aspalathoides Vahl ex DC. on Swiss albino rats by Cyclophosphamide induced myelosuppression, In vivo phagocytic activity by carbon clearance assay, Plaque forming cell (PFC) assay, flow cytometric studies for the estimation of T lymphocytes (CD3 and CD19) and Th1 cytokines (IL-2, IFN-γ and IL-4). Levamisole (50 mg/kg b.w.p.o) and Cyclophosphamide (30mg/kg b.w.i.p.) were used as immunostimulatory and immunosuppressive standard drugs. Administration of Kaempferol-5-O-β-D-glucopyranoside at the doses of 10, 20, 40, 50 mg/kg b. w. p. o. showed significant increase in the phagocytic index and in Plaque forming cell assay, and an significant protection against cyclophosphamide induced neutropenia and showed dose dependent increase in the production of T lymphocytes (CD3 and CD19) and Th 1 cytokines (IL-2, IFN-γ and IL-4). The maximum increase in the production of CD3, CD19, IL-2, IFN-γ and IL-4 at 50 mg kg b.w of the test compound was observed. The above results suggest that Kaempferol-5-O-β-D-glucopyranoside from methanol extract of aerial parts of Indigofera aspalathoides Vahl ex DC. can be further explored for the development of potent immunomodulatory drug.