Anti-Th17 and anti-Th2 responses effects of hydro-ethanolic extracts of Aframomum melegueta, Khaya senegalensis and Xylopia aethiopica in hyperreactive onchocerciasis individuals' peripheral blood mononuclear cells

Author summarySevere form of human onchocerciasis also called hyperreactive onchocerciasis (HO) is characterized by skin disorders such as dermatitis. Our previous study showed that concomitant Th2 and Th17 responses play a role in the pathophysiology of the disease. In Onchocerca volvulus endemic areas such as Togo, traditional healers (TH) used to treat this disease with medicinal plants such as Aframomum melegueta (AM), Xylopia aethiopica (XA) and Khaya senegalensis (KS). Mass drug administration including ivermectin and albendazole has been used over the past decade to control and eradicate helminths infections. Despite its successes, resistance to ivermectin has been reported and therefore, alternative drugs are urgently needed. We previously confirmed in vitro the anthelminthic effects of Aframomum melegueta (AM), Xylopia aethiopica (XA) and Khaya senegalensis (KS). However, their effect on severe onchocerciasis has not been demonstrated. Here, the anti-Th17 and anti-Th2 effects of hydro-ethanolic extracts of AM, XA and KS during HO were investigated. The data showed that in addition to their helminth-killing effects, plant-derived molecules AM, XA and KS downregulated Th2 and Th17 profiles and therefore, could be candidates for the development of new drugs not only for the treatment of helminth-induced inflammatory pathologies but also auto-immune Th2/Th17-dependent inflammatory diseases in general. Hyperreactive onchocerciasis (HO) is characterized by a severe skin inflammation with elevated Th17-Th2 combined responses. We previously demonstrated the anthelminthic activity of Aframomum melegueta (AM), Xylopia aethiopica (XA) and Khaya senegalensis (KS) used by traditional healers to treat helminthiasis in the endemic area of Togo. However, their effect on severe onchocerciasis is poorly investigated. The present study aimed to investigate the anti-Th17 and anti-Th2 effects of hydro-ethanolic extracts of AM, XA and KS during HO. Onchocerca volvulus-infected individuals were recruited in the Central region of Togo in 2018. Isolated peripheral blood mononuclear cells (PBMCs) from both generalized onchocerciasis (GEO) and HO forms were activated with anti-CD3 and anti-CD28 monoclonal antibodies in the presence or absence of the hydro-ethanolic extracts of AM, XA and KS as well as their delipidated, deproteinized and deglycosylated fractions. After 72 hours, cytokines were assayed from cell culture supernatants. Then, flow cytometry was used to investigate the effects of the extracts on cell activation, proliferation, intracellular cytokines and T cells transcription factors. The production of both Th17 and Th2 cytokines IL-17A and IL-5 were significantly inhibited upon T-cell receptor (TCR) activation in the presence of the hydro-ethanolic extracts of AM, XA and KS in HO individuals' PBMCs in vitro. AM and XA inhibited CD4(+)RORC2(+)IL-17A(+) and CD4(+)GATA3(+)IL-4(+) cell populations induction. This inhibition was not Th1 nor Treg-dependent since both IFN-gamma and IL-10 were also inhibited by the extracts. AM and XA did not interfere with T cell activation and proliferation for their inhibitory pathways. Lipid and protein compounds from AM and XA were associated with the inhibition of IL-17A. This study showed that in addition to their anthelminthic effects, hydro-ethanolic extracts of Aframomum melegueta, Xylopia aethiopica and Khaya senegalensis could downregulate both Th17 and Th2 responses and prevent the severe skin disorder observed.