Eliciting Differentiation of Human T-Cells into TH1 or TH2 Cells with Inorganic Arsenic
Loading...
Links to Files
Permanent Link
Collections
Author/Creator
Author/Creator ORCID
Date
2019
Department
Biological Sciences
Program
Citation of Original Publication
Rights
Abstract
A known carcinogen at high levels, inorganic arsenic is naturally found in groundwater and soil because of its biogeochemical cycle. Populations across the world, including in the United States, are exposed to it. However, its effects at low doses are not as well known. It has been reported that inorganic arsenic, in the form of arsenite (Asm) causes immunosuppression via inflammation. However, the exact cellular mechanism in human T-cells is not well known. This study is to develop a method by which to identify whether arsenite causes T-cells to differentiate to a T-helper I (THI) inflammatory response or a T-helper 2 (TH2) anti-inflammatory response by cytokine expression secretion analysis. The following cytokines were studied: IL-2 for cell activation, proliferation and differentiation; IFN-y for the pro-inflammatory THI response; and IL-4 for the anti-inflammatory TH2 response. Using the Jurkat cell line, the cell cultures were treated and incubated for 24 hours with I ng/mL of the activator phytohemagglutinin (PHA) and 20, 50, 100, 200, or 500 nM of arsenite in the form of arsenic trioxide (As2O3). ELISA color-change assays were used for cytokine protein secretion analysis, and endpoint RT-PCR was used for gene expression analysis. No secretion of lL-2, IFN-y, or IL-4 was detected. Because IL-2 is reported to be expressed and secreted by Jurkat cells upon activation, its gene expression was analyzed to determine whether the cells were successfully activated by PHA or if 24 hours had not been enough time for the cells to secrete the cytokine protein at detectable levels. However, due to contamination, the IL-2 gene expression analysis was inconclusive.