USDOE Human Subjects Research Database, Fiscal Year 2004

Medical University of South Carolina, Environmental Biosciences Program

Public Information Contact:

Dr. Janardan P. Pandey
Dept. of Microbiology and Immunology
Med. Univ. of SC
173 Ashley Ave.
Charleston, SC 29425

Phone: 843 792 4360
Fax: 843 792 2464
E-mail: pandeyj@musc.edu

Institutional Review Board (IRB):

Human Subject Projects:

Number of Human Subjects projects reported: 1

MUSCBP-95-6241

"Trichloroethylene Exposure and Host Genetic Factors in Autoimmune Diseases"

Project Identifier: MUSCBP-95-6241

"Trichloroethylene Exposure and Host Genetic Factors in Autoimmune Diseases"

Principal Investigator: Dr. Janardan P. Pandey, Medical University of South Carolina, Environmental Biosciences Program

Project started in: 1995
This project ended in fiscal year 2004.

Project Funding Information:

Funding for Human Subjects Research:

DOE: Environmental Management (EM)

$307,673.00 (Est.) for: Fiscal Year 2004

Information on Use of Human Subjects:

This project does not involve the use of multiple protocols/subprojects.

Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Medical University of South Carolina, Environmental Biosciences Program
Most recent approval: 12/06/02
IRB approval number: 6241
Explanation of IRB approval:
This protocol was terminated on 10/24/03 and subsequently approved as Exempt Research (#9671).

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 355
Reporting period for number of human subjects: Fiscal Year 2004

Type(s) of Human Subjects Involvement:

Collection of personally identifiable bodily materials (blood or blood products, urine, cells, tissue, teeth, organs, excreta, etc):

• Using bodily materials collected specifically for this project.

Use of personally identifiable data from questionnaires, surveys, or epidemiological studies:

• Using data collected from subjects specifically for this project.

(a. Objectives, b. Methodology, c. Ionizing Radiation, Radioactive Substances, or Chemical Substances to which human subjects are exposed, d. Involvement of Human Subjects [d.1. procedures used, d.2. risks if any])

The overall long-term goal of this project is to identify the genetic and environmental factors that contribute to the pathways to autoimmunity. In particular, experiments are underway to determine how certain genes of the immune system and those involved in the bioactivation of particular environmental toxicants interact in causing autoimmune diseases. The murine experiments in progress are designed to delineate the biological mechanisms underlying the observed association between trichloroethylene (TCE) exposure and the risk of developing Systemic Sclerosis (SSc). Apoptosis of endothelial cells is thought to be the initial pathogenic event in SSc. Experiments are in progress to determine whether molecular mimicry between certain viral and endothelial cell antigens contributes to the generation of autoantibodies that cause apoptosis of endothelial cells in SSc patients.

The specific aims of the project are: (1) to determine whether exposure of mice to TCE causes activation of microchimeric cells and the appearance of dermal inflammation and fibrosis similar to that observed in SSc patients, (2) to determine whether exposure of mice to TCE induces the production of autoantibodies associated with SSc (anti-topoisomerase I, anti-centromere, and anti-RNAP I and III) and Systemic Lupus Erthyematosus (SLE) (anti-Sm and anti-ds DNA), (3) to determine whether antibodies to human cytomegalovirus protein UL 94 produced in SSc patients crossreact with endothelial cell antigens, (4) to compare the distribution of particular genetic markers (HLA, TNF-alpha, TNF-beta, 1L-1, IL-1RA, IL-10, CTLA-4, cytochrome P450IIE1, GM, and KM) among TCE/silica exposed SSc, SLE, and myositis patients with (a) non-exposed patients and (b) non-autoimmune controls, (5) to delineate the mechanisms underlying the association between immunogenetic markers (especially HLA and GM) and SSc/SLE associated autoimmune responses, and (6) to continue investigating whether the TCE-SSc association observed in the U.S. subjects holds for other world populations as well.

All genetic markers will be characterized either by direct DNA sequencing or by the polymerase chain reaction (PCR)-RFLP methods devised by us or previously described by other investigators. Disease-specific autoantibodies and antibodies to endothelial cell and viral epitopes will be measured by Enzyme-Linked Immunosorbent Assay (ELISA), western blot, indirect immunofluorescence, double immunodiffusion, and RNA and protein immunoprecipitation assays. Synthetic peptides, recombinant proteins, or commercially available antigens will be employed in these experiments. In murine experiments, microchimeric cells in peripheral blood will be quantitated by incubation with monoclonal antibodies to H-2 molecules, which differ between BALBc/J and C57 mice, followed by FACS analysis. Paraffin embedded tissues will be used for histological assessment of collagen deposition and cellular infiltration, and also for fluorescent in situ hybridization to quantitatively measure microchimeric (male) cells using X and Y chromosome specific probes. Collagen content of hydrolyzed skin samples will be determined by a colorimetric assay. Job exposure matrices and an adapted version of expert assessment methods will be used to quantify occupational exposure to TCE and silica, respectively. Logistic regression, log-linear analyses, chi-square, and Fisher's two-tailed exact tests will be used to analyze the data, employing the SAS software (SAS Institute Inc., Cary, NC) or other programs available on the Internet.