Grantee Research Project Results
2006 Progress Report: Development of Molecular Biomarkers to Measure Environmentally Induced Immune Responses
EPA Grant Number: R832947Title: Development of Molecular Biomarkers to Measure Environmentally Induced Immune Responses
Investigators: Miller, Lisa A. , Gilliland, Frank D. , Margolis, Helene , Gern, James , Joad, Jesse , Abel, Kristina
Current Investigators: Miller, Lisa A. , Gilliland, Frank D. , Abel, Kristina , Margolis, Helene , Gern, James , Joad, Jesse
Institution: University of California - Davis
EPA Project Officer: Callan, Richard
Project Period: June 15, 2006 through June 14, 2009 (Extended to December 14, 2011)
Project Period Covered by this Report: June 15, 2006 through June 14, 2007
Project Amount: $712,423
RFA: Early Indicators of Environmentally Induced Disease (2004) RFA Text | Recipients Lists
Research Category: Human Health
Objective:
The objective of this study is to establish a panel of immune biomarkers that will be used to detect environmentally induced disease in humans, focusing on parameters of allergy and asthma. The panel of immune biomarkers will be developed by completion of the following two specific aims:
- Develop a high-throughput quantitative RT-PCR assay for leukocyte cluster of differentiation markers to measure key immune cell populations associated with allergy and asthma in small biological samples.
- Develop a high-throughput quantitative RT-PCR assay for key cytokine markers associated with allergy and asthma to measure immune cell function in small biological samples.
Progress Summary:
The goal for Year 1 was to complete 11 immune biomarkers for flow cytometry and RT-PCR. To date, we have successfully isolated and purified rhesus macaque CD3 (pan-T cell), CD4 (T helper), and CD8 (T cytotoxic) populations for mRNA analysis. Because of funding delays, molecular assessment has been limited to the following seven immune biomarker targets: CD3 epsilon, CD4 p55, CD8 alpha chain, CCL11, CCL24, CCL26 and CCL20.
To expedite the process of generating standard curves for RT-PCR analysis, we have purchased commercially prepared purified human plasmid targets. Because these data will be important for determination of key immune biomarkers for the National Children’s Study, we have decided to utilize primers and SYBR green as our methodology to rapidly assess gene expression in sample populations. This will allow us to evaluate rhesus monkey and human samples without technical delays in the design and testing of additional reagents. Further, it is likely that the National Children’s Study will redesign some of the reagents used for testing if mass spectrometry (MALDI-TOFF) equipment will be used for centralized analysis of samples collected at individual sites. Therefore, our primary goal will remain focused on providing data on specific immune biomarker targets that can be incorporated into the National Children's Study.
In addition to making some technical changes to facilitate rapid completion of the specific aims, we have shifted some of the gene targets from Year 2 to Year 1. This is primarily due to recent data obtained from rhesus monkeys that indicates an important role for the eosinophil and eosinophilic chemokines during the early stages of allergic airways disease and responsiveness to episodic ozone exposure. We have also recently discovered an important link between ozone exposure and the cytokine IL-17. As discussed in the original application, with feedback from all of the investigators on this project, we will make adjustments to our gene target list if recent findings from our work or that of others would support this change.
Future Activities:
In Year 2 of this project, we will continue work on completion of additional immune biomarker targets as defined in our original application. Our focus for this subsequent reporting period is to initiate our comparative analysis of immune biomarker expression in infant rhesus monkey samples and human asthmatic subjects.
Journal Articles:
No journal articles submitted with this report: View all 6 publications for this projectSupplemental Keywords:
RFA, Health, Scientific Discipline, PHYSICAL ASPECTS, Air, HUMAN HEALTH, particulate matter, Environmental Chemistry, Health Risk Assessment, Risk Assessments, Susceptibility/Sensitive Population/Genetic Susceptibility, Allergens/Asthma, Environmental Monitoring, Physical Processes, Health Effects, genetic susceptability, ambient air quality, atmospheric particulate matter, particulates, asthma triggers, sensitive populations, asthma, air toxics, atmospheric particles, chemical characteristics, ambient air monitoring, health risks, airborne particulate matter, asthma indices, environmental risks, exposure, second hand smoke, airway disease, airway inflammation, air pollution, aerosol composition, atmospheric aerosol particles, human exposure, airborne pollutants, inhalation, human susceptibility, allergic response, tobacco smokeProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.