In vitro effects of PCDDs/Fs on NK-like cell activity of Eisenia andrei earthworms

Hayet Belmeskine,Michel Fournier,Louise Vandelac,Sami Haddad

doi:10.4081/xeno.2012.e1

Abstract

In this study, we assessed in vitro the effects of PCDD/Fs on the NK-like cell activity in Eisenia andrei earthworms using flow cytometry for analysis. NK-like coelomocytes isolated from E. andrei and used as effectors were exposed to various concentrations of PCDDs/Fs mixture, C1 (6.25x10-3 ng 2378- TCDD/mL), C2 (12.5x10-3 ng 2378-TCDD/mL) and C3 (25x10-3 ng 2378-TCDD/mL), before adding them to human tumoral cells (K562) used as targets. We evaluated the percentage of targets lysed by Nk-like cells. The results showed a significant stimulation of the NKlike activity at C3 when PCDD/Fs were not removed from effectors before contact with targets, while no effects were noted when the effectors were washed (PCDD/Fs removed) or fixed. Assessment of the viability of the targets (K562), exposed alone and separately from effectors, to the three concentrations of PCDD/Fs, C1, C2 and C3, showed that all these concentrations were cytotoxic for K562. Results suggest that PCDD/Fs concentrations tested in this assay may be considered too low to induce suppressive effects on the immune function such as the NK-like activity in E. andrei earthworms.

Highlights

The presence of these toxic contaminants in soils may have direct harmful effects to the earth’s ecosystem.The objective of our study was to investigate, in vitro, the effects of a mixture of PCDDs/Fs on EarthwormsThe E. andrei earthworms were initially purchased from the Carolina Biological Supply (Burlington, NC, USA) and were used to estab-Earthworms play an important role in soil formation processes and in maintaining soil structure and fertility
The flow cytometry was used to study the cell-mediated cytotoxicity of earthworm NK-like cells isolated from the cœlomic cavity of E. andrei and exposed in vitro to various concentrations of PCDDs/Fs mixture
Without washing the contaminated effector cells, the in vitro PCDDs/Fs exposure significantly stimulated NK-like cell activity at C3 (25x10-3 ng 2378-TCDD/mL) compared to controls, while no effect was shown at C1 and C2 (Figure 3)

Summary

Introduction

The presence of these toxic contaminants in soils may have direct harmful effects to the earth’s ecosystem.The objective of our study was to investigate, in vitro, the effects of a mixture of PCDDs/Fs on EarthwormsThe E. andrei earthworms were initially purchased from the Carolina Biological Supply (Burlington, NC, USA) and were used to estab-Earthworms play an important role in soil formation processes and in maintaining soil structure and fertility. The presence of these toxic contaminants in soils may have direct harmful effects to the earth’s ecosystem. The objective of our study was to investigate, in vitro, the effects of a mixture of PCDDs/Fs on Earthworms. The E. andrei earthworms were initially purchased from the Carolina Biological Supply (Burlington, NC, USA) and were used to estab-. Earthworms play an important role in soil formation processes and in maintaining soil structure and fertility. They are interesting soil bio-indicators due to their ability to ingest organic and inorganic particles.[1] Currently, Eisenia andrei is recommended in standardized acute toxicity protocols[2] because of their the NK-like cell activity of E. andrei earthworm cœlomocytes to lyse human tumor cells (K562), using flow cytometry for analysis

Objectives

Methods

Results

Conclusion