Comparing [³H] thymidine LPT and CFSE assay to assess lymphocyte proliferation in beryllium-exposed sarcoidosis...

The detection of antigen specific lymphocyte responses plays a vital role in the diagnosis of various diseases. Beryllium-specific [³H] thymidine lymphocyte proliferation test (LPT) is regarded as a gold standard in identifying chronic beryllium disease (CBD) cases. Alternatively, flow cytometric based carboxyfluorescein succinimidyl ester (CFSE) assay, has several benefits as opposed to LPT, since it further permits both phenotypical characterization and functional analysis of proliferating lymphocyte subsets. The suitability of both LPT and CFSE assay to therefore detect beryllium sensitivity in a group of Be-exposed sarcoidosis patients with suspected beryllium exposure, was evaluated in this study. The clinical relevance of the test responses, expressed as stimulation indices (SI), were additionally compared on a group and individual level. Agreement in clinical interpretation of the test responses between both methods was observed in 4 out of 5 recruited patients, when considering total lymphocyte population i.e., CD3⁺ and CD19⁺-cells combined, on day 7 and with CFSE-SI >1.5, when compared with LPT-SI >2.5. Variability in responses to beryllium was additionally evaluated in Be-exposed sarcoidosis patients and compared with healthy controls. To conclude, both LPT and CFSE assay are suitable assays to detect Be sensitivity in Be-exposed sarcoidosis patients. At the same time, flow cytometric based CFSE assay has the edge over LPT in identifying the relevant proliferating lymphocyte populations. As such, when comparing two or more methods, factors that contribute to assay variability such as timepoints, lymphocyte subsets and number of replicates should always be accounted for.