EFFECTS OF COMBRETUM HEREROENSE   AND CANTHIUM MUNDIANUM   water EXTRACTS ON PRODUCTION AND EXPRESSION OF INTERLEUKIN-4

Background: Combretum hereroense and Canthium mundianum are two plants commonly used by traditional healers in the Northern region of Limpopo, South Africa for the treatment of diarrhea and inflammation. In the present study, the effects of their water extracts on the production and expression of interleukin-4 by peripheral blood mononuclear cells (PBMC’S) from HIV positive and negative individuals was evaluated.
Materials and Methods: Blood samples were collected from both HIV positive and HIV negative volunteers and were used for the purification of Peripheral blood mononuclear cells (PBMC). The PBMCs were cultured together with the water extracts after activation with phytohemagglutinin (PHA) for three days. Solid-phase sandwich ELISA (MABTECH) kit was used to detect IL-4 on un-stimulated and stimulated PBMC’S with phytohemaglutinin (PHA) and plant extracts, followed by the isolation of RNA using RNAesy Qiagen mini kit from the cells. Reverse transcriptase real time PCR was used to evaluate IL-4 gene expression by the cells.
Results: Combretum hereroense showed higher production of IL-4 at three different concentrations and a significant expression of mRNA with 4-fold amplification increase at 300µg/ml and 2-fold amplification increase at 20µg/ml. Canthium mundianum also showed increased production of IL-4 at 300µg/ml, but inhibited its production at 20µg/ml. Both extracts showed no expression at 50µg/ml. The response of the PBMCs from HIV negative individuals was more pronounced than that of HIV positive individuals who mostly increased production of IL4 at smaller concentrations unlike their HIV negative counterparts. Although in vitro studies do not necessarily predict in vivo outcomes, the plant extracts modulated the immune system by enhancing the production and expression of IL-4 in both HIV- and HIV+ individuals at different concentrations.
Conclusions: For the first time we have shown that the immunomodulatory effect of medicinal plants may depend on the clinical status of the individual. The present study revealed that the effect of the water extracts from the two plants on IL-4 expression and production is dependent on the microbiological state of the individual and is dose dependent. Further studies are needed to identify the active components in the extracts and also characterize the patients further for a better understanding of the mechanisms of action of these extracts.

Keywords
interleukin-4; peripheral mononuclear cells; HIV; ELISA; gene expression