Assessment of lymphocyte proliferation for diagnostic purpose: Comparison of CFSE staining, Ki-67 expression and 3H-thymidine incorporation
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- 作者：Jan La&scaron ; ťovička ; jan.lastovicka@lfmotol.cuni.cz ; Michal Rataj ; Jiřina Bartůňková
- 关键词：PHA ; phytohemagglutinin ; PBMC ; peripheral blood mononuclear cells ; CFSE ; carboxy fluorescein succinimidyl ester ; BrdU ; 5-bromo-2-deoxyuridine ; CPM ; counts per minute ; SI ; stimulation index ; FASCIA ; Flow-cytometric assay for specific cell-mediated immune-response in activated whole blood ; 7-AAD ; 7 amino-actinomycin D
- 刊名：Human Immunology
- 出版年：2016
- 出版时间：December 2016
- 年：2016
- 卷：77
- 期：12
- 页码：1215-1222
- 全文大小：1825 K
- 卷排序：77
文摘
The capability of lymphocytes to respond to antigenic or mitogenic stimulation is an important feature in the diagnosis of various immunodeficiencies and immune disorders. We used large cohorts of both immune compromised patients and healthy controls to measure lymphocyte proliferations by means of three methods: CFSE staining, Ki-67 expression and 3H-thymidine incorporation. The advantages and disadvantages of each method was then evaluated for use in routine clinical diagnostic. The statistical analysis was performed between the outcomes and the correlation between all three methods was computed. CFSE and Ki-67 assay correlated well with the r = 0.767, correlation between Ki-67 expression and 3H-thymidine incorporation was 0.546 and correlation between CFSE staining and 3H-thymidine incorporation was 0.337. The differences between these three methods concerning complexity, sensitivity and reliability as well as the financial aspects are discussed hereafter. CFSE and its analogues provide the cheapest and reasonable choice for measuring lymphocyte proliferation, while Ki-67 represents a more expensive, but more sensitive and robust method. The original 3H-thymidine assay does not bring any advantages and cannot compare to the competition presented by modern flow cytometric methods available today.