Title: Amelioration of Cyclophosphamide-Induced Immunosuppression after Treatment with Polyherbal Formulation Extract Running Title: Effect of PHF on CP-induced immunosuppression

The study evaluated the therapeutic potential of aqueous extract of polyherbal formulation (PHF) in cyclophosphamide (CP)-induced immunosuppression in rats. A total of 20 Wistar albino rats, 5 per group, were used. The animal was grouped into four: Normal control received 10 ml/kg BW of normal saline orally for 28 days. Negative control received 10 mg/kg BW of CP subcutaneously for three days. Treatment control received 1500 mg/kg BW of PHF orally for 28 days. The experiment group received 10 mg/kg BW of CP subcutaneously for three days, followed by immediate oral administration of 1500 mg/kg BW of PHF for 28 days. The rats were primed and challenged with 0.1 ml normal saline containing 100 μg/kg BW of KLH for delayed-type hypersensitivity (DTH) response. DTH response was assessed via paw edema using a vernier caliper. Keyhole limpet Hemocyanin (KLH)-IgG Antibody level and serum cytokines (IFN-γ, IL-4, and IL-10) were measured using sandwich enzyme-linked immunosorbent assay (ELISA). Histology of lymphoid organs was examined and reported. There is a significant difference between experimental and control groups in DTH (p<0.001), and the experimental group shows an increase in KLH-IgG antibodies compared to normal control groups. In experimental group, serum IFN-γ (p = 0.002) and IL-4 (p = 0.001) significantly increase after PHF-treatment compared to CP-treatment. Histology for normal control and experimental group shows a similar pattern. The PHF has demonstrated promising therapeutic ability in cyclophosphamide-induced immunosuppression in Wistar albino rats.

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