title: In vitro T lymphocyte proliferation by carboxyfluorescein diacetate succinimidyl ester method is helpful in diagnosing and managing primary immunodeficiencies.
abstract: BACKGROUND
  Functional studies besides routine laboratory tests for the definitive diagnosis of T lymphocyte disorders with isolated T or combined T/B-cell immunodeficiencies are important. We hereby summarized our experience with a carboxyfluorescein diacetate succinimidyl ester (CFSE)-based assay for the assessment of mitogenic T-cell proliferation responses in primary immunodeficiency (PID) patients who have not been diagnosed yet or genetically analyzed, but classified as probably having T-cell defects.
  
  
  METHODS
  Unclassified patients (n=46) and controls (n=25) were evaluated for T-cell disorders with CFSE-based assay.
  
  
  RESULTS
  CD3+blast cells after PHA-L stimulation were significantly lower in patients (31.1±28.8) than controls (67.9±8.79; P<.001). Nine patients with low and four patients with normal CD3 values had severely decreased blastic transformation. The proliferation response decreased mostly in combined immunodeficiency group. Sixteen of them had impaired proliferation responses. Appropriate molecular genetical analyses were planned after thorough evaluation of each patient.
  
  
  CONCLUSIONS
  In vitro lymphocyte cell proliferation analysis by CFSE method is a reliable and practical choice for the assessment of mitogenic T lymphocyte responses in yet unclassified PID patients for targeting further genetical analyses.
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Elif
authors.identifier: http://orcid.org/0000-0002-2887-7029
authors.initials: E
authors.lastname: Azarsiz
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Neslihan
authors.initials: N
authors.lastname: Karaca
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Birgul
authors.initials: B
authors.lastname: Ergun
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Durmuscan
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Necil
authors.initials: N
authors.lastname: Kutukculer
authors.affiliation: Department of Pediatric Immunology, Ege University, Izmir, Turkey.
authors.forename: Guzide
authors.initials: G
authors.lastname: Aksu
mesh_terms: D016022:Case-Control Studies; D049109:Cell Proliferation / Q000502:physiology*; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry; D005452:Fluoresceins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000175:diagnosis*; D007223:Infant; D008297:Male; D013388:Succinimides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013601:T-Lymphocytes / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism / Q000502:physiology
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides
keywords: T lymphocyte proliferation; carboxyfluorescein diacetate succinimidyl ester; immunodeficiency