Flow cytometric test for beryllium sensitivity.
BACKGROUND
Chronic beryllium disease (CBD) is an occupational granulomatous disorder characterized by hypersensitivity to beryllium, mediated by CD4+ T lymphocytes, and predominantly affects the lungs. In this disorder, lymphocyte proliferative responses to beryllium, measured by 3H thymidine incorporation, are used for diagnosis of CBD, for screening asymptomatic workers or former workers to detect unrecognized disease, and for surveillance as a bioassay to detect abnormal exposures. Problems with test variability and the use of radioactivity have recently led to the search for alternative methods.


METHODS
We applied a 5,6-carboxyfluorescein diacetate succinimidyl ester flow cytometric technique for measurement of mitogen- and antigen-induced T-lymphocyte proliferation to a group of beryllium-exposed sensitized individuals and beryllium-unexposed controls.


RESULTS
We detected mitogen and antigen proliferative responses in CD3+, CD4+, and CD8+ subpopulations. Phytohemagglutinin and Candida stimulated CD4+ and CD8+ T-cell responses, but beryllium appeared to stimulate only CD3+/CD4+ responses.


CONCLUSIONS
This technique may provide a sensitive, nonradioactive alternative to the traditional proliferation tests that measure beryllium sensitivity. It offers the added specificity of enabling phenotypic description of the responding cell type and may prove to be easier to standardize for clinical use.
Milovanova
Tatyana N
TN
Pulmonary, Allergy and Critical Care Division, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA.
Popma
Sicco H
SH
Cherian
Sindhu
S
Moore
Jonni S
JS
Rossman
Milton D
MD
D000395:Air Pollutants, Occupational / Q000633:toxicity*; D001607:Berylliosis / Q000276:immunology / Q000473:pathology*; D001608:Beryllium / Q000633:toxicity*; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000473:pathology; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D002908:Chronic Disease; D005434:Flow Cytometry; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D013388:Succinimides; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000473:pathology*; D013936:Thymidine / Q000378:metabolism
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000395:Air Pollutants, Occupational; D017252:CD3 Complex; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides; D001608:Beryllium; D013936:Thymidine