2000 Progress Report: Asthma Susceptibility to PM2.5EPA Grant Number: R827351C003
Subproject: this is subproject number 003 , established and managed by the Center Director under grant R827351
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: EPA NYU PM Center: Health Risks of PM Components
Center Director: N/A
Title: Asthma Susceptibility to PM2.5
Investigators: Thurston, George D. , Reibman, Joan
Institution: New York University School of Medicine
EPA Project Officer: Chung, Serena
Project Period: June 1, 1999 through May 31, 2005 (Extended to May 31, 2006)
Project Period Covered by this Report: June 1, 2000 through May 31, 2001
RFA: Airborne Particulate Matter (PM) Centers (1999) RFA Text | Recipients Lists
Research Category: Air Quality and Air Toxics , Particulate Matter , Air
The objectives of this project are to identify which particulate matter (PM) component(s) and mechanisms affect asthmatics most strongly, and to follow a cohort of non-smoker asthmatics to evaluate PM effects on their health status. The specific goals are to: (1) establish technical and operational feasibility for a combined epidemiological/clinical research study; (2) demonstrate associations between specific PM components and commonly occurring asthma exacerbations attributable to air pollution; and (3) develop hypotheses regarding the mechanisms of PM-health effects association that can be tested via toxicological studies by other researchers in the New York University (NYU)-Environmental Protection Agency (EPA) Research Center (e.g., via controlled exposure studies).
In the winter of 1999-2000, recruitment continued of patients for our cohort of adult non-smoking asthmatic subjects willing to be followed by prospective monitoring, on days following low versus high PM2.5 concentrations. Because of difficulties in the first summer (1999) in inducing sputum in asthma patients, we felt we needed to improve our induced sputum technique. Approval was obtained to induce sputum from normal volunteers, and 10 subjects were recruited and duplicate procedures performed on these subjects.
Subjects with asthma were recruited from the previous summer cohort, clinics, and local advertisements. Participants were asked to be "on call" for 1 day notice to come for four visits, two "high" and two "low" visits. These correspond to 2 day lag visits from the defined day. Subjects then underwent pulmonary function testing (PFT), blood draw, pre-medication with bronchodilator, followed by sputum induction. "High" and "low" PM days were defined based on analysis of previous data: "high" = PM10 40 micrograms (µg)/m3, while "low" = PM10 20 µg/m3.
Sputum induction was performed by increasing concentrations of hypertonic saline that were inhaled (3, 4, and 5 percent) for 7 minutes, via an ultrasonic nebulizer. Subjects underwent spirometry for measurement of forced expiratory volume in 1 second (FEV1) at the start of the procedure, and after each period of inhalation. If the FEV1 dropped 20 percent, the procedure was terminated. After each saline inhalation, subjects coughed into a sterile container. Sputum plugs were separated from saliva and examined within 2 hours. After weighing, sputum plugs were dissolved in dithiothreitol (0.1 percent) and phosphate buffered saline. The suspension was then filtered and a total nonsquamous cell count performed. Cell viability was determined by trypan blue exclusion. Cytospins were prepared, stained with Wright's stain, and a differential cell count of nonsquamous cell types was performed. Metachromatic cells were detected in preparations stained with toluidine blue. Cell pellets were also prepared for ribonucleic acid (RNA) analysis.
To date, sputum samples have been successfully collected on both normal subjects (n = 10) and from subjects with asthma (n = 11). In addition, some 44 blood serum samples were collected. This is not yet sufficient for the originally envisioned high versus low PM day comparisons. However, these samples can provide a basis for evaluating which biomarkers can be successfully used to assess PM-induced effects in the study population. For example, as shown in Figure 1, preliminary findings from several of these samples have already demonstrated the ability to detect and measure inflammatory cells in sputum samples, as well as the presence of elevated levels of eosinophils. In addition, sputum samples were analyzed for the presence of dendritic cells (CD1a+), and the quality of messenger RNA (mRNA) was tested in sputum cell pellets.
Figure 1. Percent of Total Inflammatory Cells
Overall, progress was made toward our study goals, but practical problems arose limiting our success in achieving the originally proposed results within the anticipated timeframe. The number of subjects that reliably participated was too limited, and very few days in the rainy summer of 2000 met our "high" PM pollution day criteria. Only 2 days in the summer of 2000 met the "high" pollution day criteria, as opposed to an expected 18 days. Furthermore, only 50 percent of our previous subjects agreed to return for the study. Forty subjects were screened for PFT and clinical parameters. Twenty of these subjects failed screening on PFT criteria even after modification of exclusion criteria; 13 patients agreed to participate in the screening. These factors conspired to significantly reduce the number of sample-days that could be collected.
The limitations in our ability to collect and analyze samples forced us to re-examine our results and adjust our approach in order to better work towards achieving our goals. We plan to analyze the paired sputum and serum samples that we have collected from both normal and asthma subjects to date, in order to assess those specific indicators that can be used successfully as asthma biomarkers. In particular, we propose to follow up on some recent animal studies by Dr. J. Zelikoff (NYU-EPA PM Center), which demonstrate that inhalation of concentrated ambient PM alters circulating immune blood cell profiles in a manner indicative of a stress response. Thus, we plan to analyze this limited number of sputum and serum samples for assessing acute inflammatory cytokine markers, including IL-1,6,8,10 and tumor necrosis factor. In addition, metallo-proteinases derived from inflammatory and structural cells that have been reported in serum (e.g., MMP-9 and TIMP) will be evaluated in these samples.
Based upon our findings from the already collected samples, new samples will be collected weekly on a limited number of asthma subjects during the summer of 2001 (e.g., 10 weekly, at 2 per day during Monday-Friday). This scheduled design is expected to avoid past problems experienced in trying to bring in subjects on short notice. Those cytokines that we will, by that time, have shown can be meaningfully evaluated in these samples, as well as changes in immune cellular responses (including lymphoproliferation), will be determined in both serum and sputum media. Moreover, if effects upon neutrophil emigration in human subjects are found to be similar to those previously observed by Dr. Zelikoff in her separate animal model studies, additional endpoints will be examined in the sera, potentially including soluble, tumor necrosis factor (TNF) receptor and adhesion molecules. This new approach, therefore, will utilize results from the data that we have collected to date, as well as address past data collection problems in the original study design.
Journal Articles:No journal articles submitted with this report: View all 4 publications for this subproject
Supplemental Keywords:particulate matter, PM, exposure, epidemiology, clinical, toxicology, asthma, smoker, non-smoker, sputum., RFA, Health, Scientific Discipline, PHYSICAL ASPECTS, Air, ENVIRONMENTAL MANAGEMENT, HUMAN HEALTH, particulate matter, Environmental Chemistry, Health Risk Assessment, Risk Assessments, Susceptibility/Sensitive Population/Genetic Susceptibility, Allergens/Asthma, Environmental Monitoring, Health Effects, Physical Processes, genetic susceptability, Atmosphere, Risk Assessment, ambient air quality, atmospheric particulate matter, particulates, asthma, asthma triggers, sensitive populations, air toxics, atmospheric particles, chemical characteristics, toxicology, ambient air monitoring, health risks, airborne particulate matter, ozone, asthma indices, environmental risks, exposure, second hand smoke, airway disease, airway inflammation, air pollution, aerosol composition, atmospheric aerosol particles, human exposure, airborne pollutants, inhalation, ozone monitoring, human susceptibility, allergic response, tobacco smoke, exposure assessment
Progress and Final Reports:Original Abstract
Main Center Abstract and Reports:R827351 EPA NYU PM Center: Health Risks of PM Components
Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R827351C001 Exposure Characterization Error
R827351C002 X-ray CT-based Assessment of Variations in Human Airway Geometry: Implications for Evaluation of Particle Deposition and Dose to Different Populations
R827351C003 Asthma Susceptibility to PM2.5
R827351C004 Health Effects of Ambient Air PM in Controlled Human Exposures
R827351C005 Physicochemical Parameters of Combustion Generated Atmospheres as Determinants of PM Toxicity
R827351C006 Effects of Particle-Associated Irritants on the Cardiovascular System
R827351C007 Role of PM-Associated Transition Metals in Exacerbating Infectious Pneumoniae in Exposed Rats
R827351C008 Immunomodulation by PM: Role of Metal Composition and Pulmonary Phagocyte Iron Status
R827351C009 Health Risks of Particulate Matter Components: Center Service Core
R827351C010 Lung Hypoxia as Potential Mechanisms for PM-Induced Health Effects
R827351C011 Urban PM2.5 Surface Chemistry and Interactions with Bronchoalveolar Lavage Fluid (BALF)
R827351C012 Subchronic PM2.5 Exposure Study at the NYU PM Center
R827351C013 Long Term Health Effects of Concentrated Ambient PM2.5
R827351C014 PM Components and NYC Respiratory and Cardiovascular Morbidity
R827351C015 Development of a Real-Time Monitoring System for Acidity and Soluble Components in Airborne Particulate Matter
R827351C016 Automated Real-Time Ambient Fine PM Monitoring System