USDOE Human Subjects Research Database, Fiscal Year 2000

University of Vermont

Public Information Contact:

Dr. Richard J. Albertini
Genetic Toxicology Laboratory
32 N. Prospect St.
Burlington, VT 05401-0508

Phone: 802-656-8346
Fax: 802-656-8333
E-mail: ralberti@zoo.uvm.edu

Institutional Review Board (IRB):

Human Subject Projects:

Number of Human Subjects projects reported: 1

UVT-97-DE-FG07-96ER62331

"A Novel Biomarker for Beryllium Sensitization in Humans"

Project Identifier: UVT-97-DE-FG07-96ER62331

"A Novel Biomarker for Beryllium Sensitization in Humans"

Principal Investigator: Dr. Richard J. Albertini, University of Vermont

Project started in: 1997

Project Funding Information:

Funding for Human Subjects Research: No Funding Sources Reported

Information on Use of Human Subjects:

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

Identifier or number: CHRMS 96-269

Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: University of Vermont
Most recent approval: 01/19/00
IRB approval number: 01XB
Explanation of IRB approval:
No cost one-year extension.

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 2
Reporting period for number of human subjects: Fiscal Year 2000 (10/1/99-9/30/2000)

Type(s) of Human Subjects Involvement:

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

• Using cells cultured in a laboratory.
• Using bodily materials collected 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])

This research project will determine the T-cell (TCR) gene usages of beryllium reactive T-lymphocytes isolated directly from the peripheral blood of individuals exposed at a U.S. Department of Energy site. The objective is to develop a sensitive and novel biomarker for identifying early human sensitization to environmental beryllium. This is a collaborative project involving the Genetic Toxicology Laboratory of the University of Vermont and both the Center for Epidemiological Research and the scientific staff of the Cytogenetics Program at the Oak Ridge Institute for Science and Education (ORISE). The >2000 beryllium exposed workers who have been contacted for participation in the ORISE study "Follow-up of Beryllium Workers at the Y-12 Plant/Efficacy of the Peripheral Blood Lymphocyte Proliferation (LPT) and Other Non-Invasive Procedures for Diagnosis of Chronic Beryllium Disease" will provide the pool of potential participants for the proposed study. Additional subjects may be obtained from the National Jewish Hospital, Denver, CO.

Beryllium reactive T-lymphocytes will be directly isolated from peripheral blood using a novel antigen-independent method of surrogate selection for in vivo arising hypoxanthine phosphoribosyltransferase (hprt) mutants as representatives of clones that are undergoing chronic proliferation. The T-cells undergoing chronic proliferation in beryllium sensitized individuals will be enriched for beryllium reactive cells. The TCR beta gene usage of these T-cell isolates will be determined and their junctional (CDR3) regions sequenced. Beryllium reactive T-cell clones will also be recovered following in vivo beryllium stimulation of peripheral blood lymphocytes from these same individuals and the TCR gene CDR3 region sequences similarly determined. The TCR beta genes used by the beryllium reactive isolates and their CDR3 region sequences will be compared within (in vivo vs. in vitro derived) and among individuals with attention to kinds and durations of beryllium exposure and HPA DPB Glu 69 status. A method for quantitating total body loads of these antigen reactive T-cells in individuals will be developed using quantitative polymerase chain reaction (Q-PCR) amplification of specific TCR gene sequences. Successful achievement of this overall objective will permit future studies aimed at the elucidation of the immunological mechanisms underlying sensitization, the comparison of cells involved in pulmonary and systemic sensitization and the definition of potential targets for immunotherapy. A method of mass beryllium stimulated PBL cultures from study subjects will be used to identify T-cell receptors (TCR) used by individuals to recognize the beryllium antigens. A QPCR method will be used for TCR identification.

Blood will be obtained with informed consent from persons previously exposed to beryllium. Cells isolated from this blood will be cultured in vitro. There is essentially no risk except for possible bruising at the site of blood drawing. This protocol has University of VT IRB approval and informed consent is obtained from all individuals studied. The results of these studies are confidential.