Abstract
BackgroundRhaphidophora korthalsii (Araceae) is a root-climber plant which has been widely used in Chinese traditional medicine for cancer and skin disease treatment. Previous reports have recorded its immunomodulatory effects on mice splenocyte and human peripheral blood. This study investigated the potential immunostimulatory effect of Rhaphidophora korthalsii on human PBMC enriched NK cell.MethodsPBMC was exposed to various concentrations of R. korthalsii extract and the T and NK cell population in the control and extract treated PBMC were identified by immunophenotyping. Intracellular perforin and granzyme B expressions were detected by flow cytometry and extra-cellular Granzyme B, IFN-γ and TNF-α production in the isolated NK cells were determined by ELISA. The cytotoxicity of effector NK cell towards target K562 cell was assessed by CytoTox 96 assay.ResultsRhaphidophora korthalsii methanol extract significantly increased PBMC NK cell population and intracellular perforin and granzyme B expressions. Moreover, the extract also enhanced the secretion of IFN-γ and TNF-α which subsequently enhanced the cytotoxicity of NK cell against the NK sensitive target K562 cell line. NK cell enriched with extract treated PBMC showed better activation than NK cell directly treated with the extract.ConclusionOur findings indicated a potential IL-2 free immunotherapy through direct and indirect R. korthalsii stimulation on NK cell activation.
Highlights
Rhaphidophora korthalsii (Araceae) is a root-climber plant which has been widely used in Chinese traditional medicine for cancer and skin disease treatment
Reagents and chemicals Fluoroisothiocyanate (FITC)- labeled CD16, CD3, perforin, granzyme B, and phycoerytherin (PE)-labeled CD56 and IL-2 neutralizing MQ1-17H12 monoclonal antibodies, GolgiStopTM solution and Perm/WashTM solution were purchased from BD, USA; isotype control mouse IgG1 and FITC goat anti-rabbit Ig were purchased from Abcam, USA; Enzyme Link Immunosorbent Assay (ELISA) kit for IFN-γ, TNF-α and Granzyme B cytokines from Bender MedSystems, Austria; NK cell and T cell isolation kits from Milteny Biotech, Germany; human recombinant human Interleukin 2, trypan blue solution, DMEM were brought from Sigma, USA
Searching for novel immunomodulator which can work synergistically with IL-2 to reduce or replace its working dosage has been proposed from time to time and this may contribute to the success of adoptive immunotherapy in cancer treatment
Summary
Rhaphidophora korthalsii (Araceae) is a root-climber plant which has been widely used in Chinese traditional medicine for cancer and skin disease treatment. This study investigated the potential immunostimulatory effect of Rhaphidophora korthalsii on human PBMC enriched NK cell. The focus on the study of biological activities such as immunostimulatory effect from plant extracts based on ethnopharmacology has been revived all over the world due to the low or absence of toxicity, complete biodegradability, Rhaphidophora korthalsii, Schott. The extract had showed to exert immunomodulatory effect on mice splenocyte and human peripheral blood mononuclear cell (PBMC) proliferation, increase of the NK cell population, cytokines secretion and cytotoxicity in vitro and in vivo [8,9,10,11]
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