Publication: Identifying The Effect Of Biosynthesized Agnp From Garcinia Atroviridis Aqueous Extract On Induced T-regulatory Cells From Non-obese Resistance (Nor) Mouse Model
Loading...
Date
2018-08
Authors
MD Yusof, Nur Azirah
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
CD4+CD25+ Foxp3+ T regulatory cells (Tregs) are important for maintenance of immune homeostasis. Thus, harnessing their function as immune modulator may be coupled with the rapid advancement of nanotechnology development. In the current study, biosynthesized silver nanoparticle (AgNP) from aqueous extract of Garcinia Atroviridis (GA) was used to identify its synergistic effect on modulatory properties of induced Treg (iTreg) cells from Non-Obese Resistance (NOR) Mice Model. Conventional CD4+ T cells were isolated using magnetic separation from NOR mice spleen and then cultured. Next, 1x105cells/mL were stimulated with anti CD3/CD28, TGF-B and IL-2 to induce CD4+Foxp3+ cells (iTreg). The cells were treated with or without GA-AgNP and were cultured for 120 hours. Expression of extracellular and intracellular markers were evaluated by using fluorochrome dye to detect CD4+, Foxp3,IL-17R, ICOS, TIGIT, P38, STAT5 and ERK1/2. Cells were analysed using BD FACS DIVA Cell analysis system. Control group was set as CD4+IL17Rhigh and stimulated with anti-CD3/CD28 and IL-2 only. Treated groups were designated as following: Group 1. anti-CD3/CD28, IL-2, TGF- ẞ Group 2. anti-CD3/CD28, IL-2, TGF- ẞ, GAAgNP and 3. anti-CD3/CD28, IL-2, GA-AgNP. Current results showed that GA-AgNP modulated the expressions of several molecules on isolated CD4+IL17Rhigh T cell population. Levels of CD4+ expression were significantly reduced in Group and Group 3 which were treated with GA-AgNPs as compared to control group. Similarly, Group 1 expressed almost similar patterns of molecular expression with control group. In contrast, transcription factor Foxp3 was not significantly induced in all groups as compared to control group. The expression of ICOS and TIGIT markers in CD4+IL17Rhigh cells in all groups remained unchanged as compared to control group.In contrast, transcription factor Foxp3 in all groups was not significantly induced compared to control group. The phosphorylation of p38 protein and ERK1/2 of MAPК pathways did not significantly detected in all groups of cells as compared to control group. None of the cell groups showed significant STAT5 phosphorylation level compared to control group. These findings may suggest GA-AgNP in modulating CD4+IL-17Rhigh cell population and did not significantly induce the formation of CD4+Foxp3+ cells. Thus, the addition of GA-AgNP in TGF-B/IL2 environment may trigger the plasticity of CD4+ cells into different population. Further studies are necessary to identify the molecular mechanisms involved. Current findings may serve as a basis for immunotoxicity study of nanomaterial used in therapeutic bio-products.
Description
Keywords
immune modulator , immune regulation , NOR , GA-AgNP , iTreg