Innate Immune Response of an Aquatic Vertebrate Model to Manufactured Nanoparticles Assessed Using Genomic MarkersEPA Grant Number: R833319
Title: Innate Immune Response of an Aquatic Vertebrate Model to Manufactured Nanoparticles Assessed Using Genomic Markers
Investigators: Klaper, Rebecca , Goetz, Frederick , Chen, Jian
Institution: University of Wisconsin Madison
EPA Project Officer: Hahn, Intaek
Project Period: April 1, 2008 through April 15, 2011
Project Amount: $398,810
RFA: Exploratory Research: Nanotechnology Research Grants Investigating Environmental and Human Health Effects of Manufactured Nanomaterials: a Joint Research Solicitation-EPA, NSF, NIOSH, NIEHS (2006) RFA Text | Recipients Lists
Research Category: Health Effects , Nanotechnology , Health , Safer Chemicals
The innate immune system is one of the first physiological systems to interact with foreign materials and, therefore, will be key to understanding how organisms will be affected by exposure to nanomaterials. Recent studies have indicated that the innate immune system of fish responds to certain pathogen patterns differently than mammals. Therefore, the response of the mammalian immune system may not necessarily be representative of the immune reaction of aquatic vertebrates such as fish. Past cellular studies have concentrated on general cytotoxicity.
The overall objective of this project is to assess the innate immune reaction of an aquatic model, the rainbow trout, to manufactured nanomaterials of varying chemistries at levels not inducing cellular toxicity. This research will create a mechanism with which to test other nanomaterials, provide data to support ecological risk assessments, and ultimately inform decisions as to which materials will be the safest to industrialize and use with respect to aquatic environments. Our hypothesis is: nanomaterials of dissimilar chemical composition will stimulate different patterns of trout macrophage gene expression, and nanomaterials of similar chemical characteristics (e.g. charge, shape, functional group) may be grouped with respect to their bioactivity, expressed as a particular gene response pattern. Specifically, the chemical properties of nanomaterials will impact the genomic response of the immune system: nanomaterials of dissimilar chemical composition will stimulate different patterns of macrophage gene expression and the response will be dose-dependent.
A range of water-soluble C60 and carbon nanotubes with different chemical compositions and surface chemistries will be synthesized and tested for their effects on trout macrophages. A trout primary macrophage cell culture system will be used to determine the 1) dose versus cell viability for each synthesized nanomaterial type, 2) level of expression (by quantitative PCR) of marker genes associated with inflammatory, antiviral, and anti-inflammatory responses with respect to nanomaterial dose at levels that have no deleterious effect on cell viability, and 3) global patterns of gene expression for those materials that cause significant changes in marker genes using custom trout immune microarrays.
Methods developed here will improve risk assessment by creating a mechanism to test other nanoparticles prior to commercial release. The goal of this project will be to help identify nanomaterials with the least negative environmental impact for environmentally conscious manufacturing. Risk managers will use this data to identify particles for restricted release to limit harm to aquatic species.