title: Assessment of lymphocyte proliferation for diagnostic purpose: Comparison of CFSE staining, Ki-67 expression and3H-thymidine incorporation.
abstract: 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 and3H-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 and3H-thymidine incorporation was 0.546 and correlation between CFSE staining and3H-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 original3H-thymidine assay does not bring any advantages and cannot compare to the competition presented by modern flow cytometric methods available today.
authors: Department of Immunology, 2nd Medical School and University Hospital Motol, Prague, Czech Republic. Electronic address: jan.lastovicka@lfmotol.cuni.cz.
authors: Jan
authors: J
authors: Lašťovička
authors: Department of Immunology, 2nd Medical School and University Hospital Motol, Prague, Czech Republic.
authors: Michal
authors: M
authors: Rataj
authors: Department of Immunology, 2nd Medical School and University Hospital Motol, Prague, Czech Republic.
authors: Jiřina
authors: J
authors: Bartůňková
mesh_terms: D049109:Cell Proliferation; D002478:Cells, Cultured; D015331:Cohort Studies; D005434:Flow Cytometry; D005452:Fluoresceins; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D016867:Immunocompromised Host; D019394:Ki-67 Antigen / Q000378:metabolism; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D015166:Monitoring, Immunologic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013388:Succinimides; D013936:Thymidine
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D019394:Ki-67 Antigen; D013388:Succinimides; D013936:Thymidine
keywords: (3)H-thymidine incorporation; CFSE staining; Flow cytometry; Ki-67 expression; Lymphocyte proliferation; Phytohemagglutinin