Comparative analysis between CFSE flow cytometric and tritiated thymidine incorporation tests for beryllium sensitivity.
BACKGROUND
In this study, we evaluated alternative possibility for CFSE beryllium flow cytometric test against beryllium blood lymphocyte proliferation test (BeLPT) as a standard radioactive clinical screening method to identify sensitization to beryllium.


METHODS
Delta PD (the ratio of divided cell population to the total number of cells with subtracted counts of unstimulated cells) of specific beryllium-induced pathogenic CD3+ CD4+ T-lymphocytes and stimulation index (SI) in CFSE proliferation test was compared with delta counts per minute (mean test CPM minus mean control CPM) and SI in radioactive blood BeLPT.


RESULTS
Comparison analysis of CFSE and BeLPT demonstrated excellent agreement between delta PD and delta CPM (kappa = 0.845, P << 0.0001). We determined 6.8% positive subjects in the beryllium-exposed, Be-LPT-negative group. The decreased mean difference of these indexes to percentage of average and the long tail in the plot reflects increased sensitivity. CFSE/CD4+ T-cell proliferation assay has 100% specificity, significantly higher sensitivity and efficiency than BeLPT.


CONCLUSIONS
Both delta PD, measured by the precursor frequencies method in CFSE assay and delta CPM, defined by tritiated thymidine in BeLPT, can be used for the enumeration of beryllium specific CD4+ T-cell proliferation and may substantially improve the quality of the early diagnosis of beryllium hypersensitivity.
Milovanova
Tatyana N
TN
Department of Pathology and Laboratory Medicine, Flow Cytometry and Cell Sorting Facility, Abramson Cancer Center, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA. tmilovan@mail.med.upenn.edu
D001607:Berylliosis / Q000175:diagnosis* / Q000276:immunology / Q000503:physiopathology; D001608:Beryllium / Q000009:adverse effects*; D001681:Biological Assay / Q000379:methods* / Q000592:standards; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D042241:Early Diagnosis; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis* / Q000276:immunology / Q000503:physiopathology; D011237:Predictive Value of Tests; D013194:Staining and Labeling / Q000379:methods; D013234:Stem Cells / Q000187:drug effects / Q000276:immunology; D013936:Thymidine; D014316:Tritium
D014316:Tritium; D001608:Beryllium; D013936:Thymidine