Immune System Responses Related to Environmental Uranium Exposure – DiNEH Project Results

1. Immune System Responses Related to Environmental Uranium Exposure – DiNEH Project Results E. Erdei & J. Lewis University of New Mexico, Health Sciences Center, College of Pharmacy, Community Environmental Health Program Navajo Nation Human Research Review Board Conference Window Rock, AZ November 16, 2011

3. Environmental Legacy Exposures Increase the Likelihood of Several Diseases: Autoimmune Disease Figures below show similar increases in risks for autoimmune disease (self-reported) based on an increase from 1 to 2 activities

4. DiNEH Survey Responses

5. Goal of this portion of the study • Direct response to community members’ requests for research on immune system function during the capacity building and environmental risk evaluation work • Address possible pathways within the human body in association with environmental uranium and other heavy metal exposures • Find early indicators of health effects

6. DiNEH biological sample collection • DiNEH project participants from 20 chapters • Samples collected from 267 individuals • A subset has been analyzed for immune biomarkers (N=65) • Early markers, showing alterations in immune cell distribution and activity

7. Flow cytometry measurements • Lymphocyte subpopulations from whole blood samples. • Becton Dickinson Simultest IMK Plus lymphocyte kit was used. • 6 cell populations were measured: • T cells (CD3+), T helpers (CD4+), T suppressors (CD8+); • B cells (CD19+); • HLA-DR+ cell activation in T cells and • B cells and other cell types; NK cells (CD3-/CD16+/CD56+).

8. UNM HSC Core Facility • Director: Dr. Bruce Edwards • Flow cytometry machine • FACScan 2 lasers, 2-color simultaneous detection of lymphocytes • Membrane markers, CD coding

9. Flow cytometry results I.

10. Flow cytometry results II. • Increased percentage of activated T cells • Decreased percentage of activated B cells • Decoupling of T cell and B cell activities suggest altered immune response among this subset of participants • Can lead to lower production of protective antibodies • Preliminary interpretation of data; more complex modeling incorporating other variables pending

11. Serum cytokines • Cytokines are small molecular weight proteins produced by immune cells and other cells through the human body • Their role is to promote communication, activation processes in the immune system • Cytokine productions show immune status and disease developmental pathways

12. Serum cytokine measurements • Applied xMAP multiplexing technology • UNM HSC Core Facility- Luminex 100™ detection (96-well format) • Detection of 10 human serum cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, INF-γ, TNF-α, and GM-CSF) high sensitivity assay • Uses only 50 μl of serum sample/ participant

13. Serum cytokine concentrations related to environmental uranium waste exposures

14. Serum cytokine concentrations (pg/ml) related to environmental uranium waste exposures (N=47) • Percent of variance by exposure for IL-4 and IL-1ß suggests a potential inflammatory process

15. Conclusions Based on our preliminary analysis • DiNEH participants with increased exposure to uranium waste had increased number of activated T cells and decreased activity of B cells and other antigen-producing cells • If consistent w/ other modeling results (e.g. water source) – indication of the decoupling of the immune response • Alterations in cytokine production indicative of the presence of an inflammatory response

16. Future directions • These results are based on our preliminary analyses – work in progress • To continue to complete immune assays for entire sample set • Further modeling works will allow us more detailed evaluation of suggestive inflammatory response • In connection with other pathway analyses –cardiovascular process

17. NIEHS, EPA and UNM for financial support Community Advisory Board Ed Carlisle, Jay DeGroat, Herbert Enrico, Thomas Manning,Sr., Lynnea Smith, Jean Whitehorse, UNM-HSC Community Environmental Health Program & Clinical and Translational Science Center Johnnye L. Lewis, PhD; Miranda Cajero, BCH; Matthew Campen, PhD; Jeremy DeGroat; Mallery Downs, RN; Eszter Erdei, PhD; Molly Harmon; Gabriel Huerta, PhD; Curtis Miller; Bernadette Pacheco; Glenn Stark; Mary Woodruff; research nursing support Crownpoint Service Unit, I H S Virgil Davis Navajo Area IHS Lisa Allee, CNM; John Hubbard; Ryan Johnson, MD; Doug Peter, MD UT-Houston Nephrology Donald Molony, MD Southwest Research Information Center Chris Shuey, MPH, Sarah Henio-Adeky, Teddy Nez, Sandy Ramone Students Jamie deLemos, PhD – Tufts Univ. Christine George – Stanford Univ. Tommy Rock, MA, UNM Health Policy Student Christine Samuel-Nakamura, PhD Candidate, UCLA Dartmouth Ben Bostick, PhD University of Arizona Cancer Center & Northern Arizona University, NACRP Jani Ingram, PhD, Margaret Briehl, PhD USEPA Region IX Harry Allen, Rich Bauer, Clancy Tenley State of New Mexico Diagnostic Laboratory Navajo Nation EPA Air Quality Division, Public Water Supply Supervision Program, Superfund Program Navajo Nation Division of Health Former Contributors: Bess Seschillie, Bernice Norton, Jerry Elwood, Harrison Gorman, Harris Arthur (in memoriam), Alta McCabe, Margaret Menache, PhD, Alexis Kaminsky, PhD; Eastern Navajo Health Board Thanks to the many others who’ve contributed DiNEH Acknowledgements

18. Project funding support • DiNEH project – supported through the following grants • NIEHS, RO1 ES014565; R25  ES013208; P30 ES-012072; • USEPA/ERRG pass through contract; with support from DHHS/NIH/NCRR #1UL1RR031977-01

19. Questions? THANK YOU!