Detection of beryllium sensitivity using a flow cytometric lymphocyte proliferation test: the Immuno-Be-LPT.
Measurement of lymphocyte proliferation to detect hypersensitivity to beryllium (Be-LPT) in vitro is done presently using a method based on tritiated thymidine incorporation. Although this method is sensitive it gives no information on cell viability or responding lymphocyte subsets. We have developed reliable and simple flow cytometric assays for lymphocyte proliferation testing (Immuno-Be-LPT) by combining immunophenotyping with bromodeoxyuridine (BrdU) incorporation or DNA content using propidium iodide (PI) or 4'6'-diimidazolin-2-phenylindole (DAPI). Evaluation of beryllium-induced lymphocyte proliferation in blood cells from seven patients with chronic beryllium disease (CBD) and 120 beryllium workers by both the Bc-LPT and the Immuno-Be-LPT showed agreement between the tests. The Immuno-Bc-LPT provided additional information about the specific type of lymphocytes responding. CD4+ lymphocytes proliferated in response to beryllium in blood samples from all seven CBD individuals and CD8+ lymphocytes proliferated in six of the seven. Four beryllium workers without CBD had positive responses to beryllium primarily in the CD8+ cells. The use of the individual's own plasma supported a greater beryllium or tetanus-induced proliferation of CD4+ lymphocytes when compared to commercial human serum. The response of CD4+ lymphocytes measured in the Immuno-Be-LPT may provide a new marker for the diagnosis of CBD.
Farris
G M
GM
Cytometry Group, BioScience Division, Los Alamos National Laboratory, NM 87545, USA.
Newman
L S
LS
Frome
E L
EL
Shou
Y
Y
Barker
E
E
Habbersett
R C
RC
Maier
L
L
Smith
H N
HN
Marrone
B L
BL
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000096:biosynthesis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D001608:Beryllium / Q000633:toxicity*; D015415:Biomarkers; D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002908:Chronic Disease; D004247:DNA / Q000096:biosynthesis / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D037181:Lectins, C-Type; D008027:Light; D008214:Lymphocytes / Q000187:drug effects*; D009784:Occupational Diseases / Q000276:immunology; D010641:Phenotype; D012542:Scattering, Radiation; D006377:T-Lymphocytes, Helper-Inducer / Q000187:drug effects / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects / Q000276:immunology; D013745:Tetanus Toxoid / Q000494:pharmacology; D000099192:CD69 Antigens
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D001608:Beryllium; D015415:Biomarkers; D001973:Bromodeoxyuridine; D004247:DNA; D005456:Fluorescent Dyes; D037181:Lectins, C-Type; D013745:Tetanus Toxoid; D000099192:CD69 Antigens