Grantee Research Project Results
2007 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, 2007 through June 14,2008
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.
Approach:
In this proposal, we will develop a panel of oligonucleotide primers and probes for real-time quantitative PCR analysis of immune cell phenotype and function. The significance of each immune biomarker in the context of environmentally induced allergic airways disease will subsequently be determined by evaluating peripheral blood and airway cells obtained from an infant rhesus monkey model of environmentally induced asthma.
Progress Summary:
The original goal for year 2 was to have a total of 19 immune biomarkers completed for RT-PCR. Thus far, we have completed development and testing 24 immune biomarkers that are cross-reactive for rhesus monkey and human targets. Of the original list for the complete three year project, we have substituted IL-17, CCL24, and CCL20 for CD23 and Fc Epsilon receptor I targets. These new targets were selected in order to focus our efforts on T cell and eosinophil mediated responses, based on recent findings in the infant rhesus monkey. For our final year of funding, we will complete reagent development and testing for eosinophil cationic protein.
For the first specific aim in year 1, we successfully isolated and purified rhesus macaque CD3 (pan-T cell), CD4 (T helper), and CD8 (T cytotoxic) populations for mRNA analysis. Our goal was to determine if RT-PCR analysis could be used to accurately measure numbers of specific T cell populations in a peripheral blood sample. Initially, our plan was to isolate purified CD3, CD4, and CD8 cell populations from a small pool of monkeys; standard curves would be established for subsequent whole blood analysis. However, we discovered that mRNA from purified T cell populations did not correlate well with actual cell numbers as measured by flow cytometery. In contrast, mRNA from whole blood preparations do correlate well with cell numbers, as measured by flow cytometry. Although we do not know why there is a discrepancy between purified vs. whole blood populations, we suspect that the purification procedures may contribute to mRNA degradation, thereby altering the interpretation of cell numbers. To circumvent this issue, we have subsequently focused our efforts on whole blood analysis, using values obtained by flow cytometry to correlate cell number with mRNA levels.
In addition to development of biomarkers, we have begun to evaluate peripheral blood samples collected from an ongoing infant rhesus monkey study, in which animals are exposed to house dust mite allergen and ozone. Surprisingly, our preliminary analysis indicates that interferon gamma mRNA copy numbers in whole blood samples are much higher than IL-4 mRNA copy numbers. In addition, we have observed a striking increase in interferon gamma mRNA levels at approximately three months of age, suggesting a period of significant developmental shifts within the immune system.
Expected Results:
This study will generate a panel of sensitive molecular biomarkers to measure environmentally induced changes in systemic and local immune responses within small biological samples. Once tested and characterized, these reagents can be immediately incorporated as a part of the National Children’s Study to investigate how environmental exposure to air pollutants can modulate the immune system during early childhood development.
Future Activities:
In Year 3 of this project, we will complete the development of our proposed panel of immune biomarkers and focus our work on comparative analysis of immune biomarker expression in infant rhesus monkey samples and human asthmatic subjects.
Journal Articles on this Report : 1 Displayed | Download in RIS Format
| Other project views: | All 6 publications | 4 publications in selected types | All 4 journal articles |
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Chou DL, Gerriets JE, Schelegle ES, Hyde DM, Miller LA. Increased CCL24/eotaxin-2 with postnatal ozone exposure in allergen-sensitized infant monkeys is not associated with recruitment of eosinophils to airway mucosa. Toxicology and Applied Pharmacology 2011;257(3):309-318. |
R832947 (2007) R832947 (2009) R832947 (Final) |
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Supplemental Keywords:
immunology, ozone, indoor air, health effects, susceptibility, biomarkers, allergy, asthma, RFA, Health, Air, Scientific Discipline, PHYSICAL ASPECTS, HUMAN HEALTH, Susceptibility/Sensitive Population/Genetic Susceptibility, Health Risk Assessment, Physical Processes, Risk Assessments, particulate matter, genetic susceptability, Environmental Chemistry, Allergens/Asthma, Health Effects, Environmental Monitoring, sensitive populations, health risks, chemical characteristics, airway inflammation, asthma indices, asthma triggers, asthma, aerosol composition, airborne particulate matter, human exposure, ambient air monitoring, environmental risks, second hand smoke, atmospheric particles, atmospheric aerosol particles, particulates, exposure, atmospheric particulate matter, airborne pollutants, allergic response, ambient air quality, air pollution, human health risk, human susceptibility, air toxics, inhalation, airway disease
Progress 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.