Michael Thomas

Introduction
Toll-like receptors (TLRs) are trans-membranous proteins characterised within the mammalian innate and adaptive immune responses [1] . TLR activation via extracellular lipopolysaccharides (LPS) initiates a pro-inflammatory cascade via the NF- K B transcription pathway resulting in the secretion of cytokine Interleukin -8 [2] and subsequent macrophage proliferation [3] allowing TLR2 ligation to be used as a indicator of immuno -modulation. Tropical plants contain highly evolved active metabolites that can provide novel treatments against many prevailing pathogens [4] , this study assess the immuno -modulatory effects of crude M.griffiana extracts derived from the Malaysian rainforest.

Materials and Methods

H E K 293T L R + Tr a n s f e c t i o n TLR2 protein was transfected into HEK 293 cell lines (HEK+) and isolated from TLR2 negative HEK 293 line (HEK-).
Cell Culture Cell lines were cultured (37 o C, 5% CO 2 ), passaged twice a week and maintained between 60-90% confluence. M.griffiana Induced HEK Cytotoxicity 48h exposure of crude M.griffiana extract preceded a manual cell viability count using Trypan Blue. Pro-inflammatory TLR2 Ligation Interleukin-8 characterisation utilised an IL-8 specific ELISA protocol in both cell lines (48h exposure).

Results
M.griffiana Induced HEK Cytotoxicity HEK 293 TLR- cell lines were exposed to crude M.griffiana plant extract for 48h.
Cell viability was not affected in response to the plant extract (P=0.65) (Fig1.1). M.griffiana does not induce cytotoxicity in HEK 293 cell lines.

Pro-inflammatory TLR2 Ligation HEKTLR- and HEKTLR+ cell lines were exposed to crude M.griffiana plant extract for 48h. Interleukin-8 was not expressed at any extract concentration within HEK- and HEK+ cell lines (Fig1.2).

Statistical Analysis Data was analysed using Anderson-Darling, ANOVA: Single Factor and Fishers LSD analysis.

M.griffiana does not induce pro-inflammatory TLR2 ligation in HEK 293 cell lines.

Conclusion
The potential for the discovery encompassing only 1% of the tumorigenesis microenvironment [6] , in tumour stroma of novel pharmaceuticals in tropical rainforests has long been established. To date, fifty ph armaceutical products have been developed total species available in these environments [4] .TLR2 mediated activation has been shown by a number of studies to be related to the the occurrence of a pro -inflammatory response inducing plant extract will open a gateway to potential pharmaceuticals in the t reatments emphasising the need for further research into the use of tropical flowering plants as a novel pharmaceutica l resource [5] .

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