An in vitro evaluation of epigallocatechin gallate (eGCG) as a biocompatible inhibitor of ricin toxin

The catechin, epigallocatechin gallate (eGCG), found in green tea, has inhibitory activity against a number of protein toxins and was investigated in relation to its impact upon ricin toxin (RT) in vitro. The IC50 for RT was 0.08 +/- 0.004 ng/mL whereas the IC50 for RT + 100 mu M eGCG was 3.02 +/- 0.572 ng/mL, indicating that eGCG mediated a significant (p < 0.0001) reduction in ricin toxicity. This experiment was repeated in the human macrophage cell line THP-1 and IC50 values were obtained for RT (0.54 +/- 0.024 ng/mL) and RT + 100 mu M eGCG (0.68 +/- 0.235 ng/mL) again using 100 mu M eGCG and was significant (p = 0.0013). The documented reduction in ricin toxicity mediated by eGCG was found to be eGCG concentration dependent, with 80 and 100 mu g/mL (i.e. 178 and 223 mu M respectively) of eGCG mediating a significant (p = 0.0472 and 0.0232) reduction in ricin toxicity at 20 and 4 ng/ml of RT in Vero and THP-1 cells (respectively). When viability was measured in THP-1 cells by propidium iodide exclusion (as opposed to the MTT assays used previously) 10 ng/mL and 5 ng/mL of RT was used. The addition of 1000 mu M and 100 mu M eGCG mediated a significant (p = 0.0015 and <0.0001 respectively) reduction in ricin toxicity relative to an identical concentration of ricin with 1 mu g eGCG. Further, eGCG (100 mu M) was found to reduce the binding of RT B chain to lactose-conjugated Sepharose as well as significantly (p = 0.0039) reduce the uptake of RT B chain in Vero cells. This data suggests that eGCG may provide a starting point to refine biocompatible substances that can reduce the lethality of ricin. (C) 2016 The Authors. Published by Elsevier B.V.