title: Measurement of T-cell CD69 expression: a rapid and efficient means to assess mitogen- or antigen-induced proliferative capacity in normals.
abstract: We have analyzed the expression of the activation antigen CD69 on normal human T cells by flow cytometry following stimulation with mitogens and recall antigen. These data were compared to parallel studies assessing the proliferative response using the 3H-thymidine (3H-TdR) incorporation assay. Three different mitogens (PHA, ConA, and CD2/CD2R) induced maximal expression of CD69 at 24 h, which remained stable throughout the 72 h culture period. The mitogen-stimulated cells initiated DNA synthesis as determined by the 3H-TdR assay (72 h) while nonstimulated cells failed to upregulate CD69 or incorporate 3H-TdR. We next compared T cell CD69 expression (n = 12) following stimulation with either CD2/CD2R (5 micrograms/ml) or the recall antigen, tetanus toxoid (1:1500). Cell proliferation was determined by the 3H-TdR assay at 72 h (CD2/CD2R) or 120 h (tetanus toxoid). Evaluation of CD69 expression at 6 h predicted CD2/CD2R but not tetanus responder status as defined by 3H-TdR incorporation. However, when 4 known tetanus responders (3H-TdR) were evaluated over time, it was found that at 48 h the fluorescence intensity (of CD69) on tetanus-stimulated CD3+ cells increased markedly compared with nonstimulated cells (range of increase 43-850%). One individual whose cells failed to respond to tetanus toxoid (3H-TdR and CD69) did respond normally to CD2/CD2R. These observations suggest that flow cytometric evaluation of T cell CD69 expression following mitogen (6 h) or antigen (48 h) stimulation may provide an accurate screen of T-cell responsiveness in normals.
authors.affiliation: Laboratory of Host Defenses, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: M
authors.initials: M
authors.lastname: Mardiney
authors.forename: M R
authors.initials: MR
authors.lastname: Brown
authors.forename: T A
authors.initials: TA
authors.lastname: Fleisher
mesh_terms: D015703:Antigens, CD / Q000032:analysis*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis*; D018801:CD2 Antigens / Q000276:immunology*; D002455:Cell Division; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007700:Kinetics; D037181:Lectins, C-Type; D008934:Mitogens / Q000494:pharmacology*; D010835:Phytohemagglutinins / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D013745:Tetanus Toxoid / Q000276:immunology*; D013997:Time Factors; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018801:CD2 Antigens; D003208:Concanavalin A; D037181:Lectins, C-Type; D008934:Mitogens; D010835:Phytohemagglutinins; D013745:Tetanus Toxoid; D000099192:CD69 Antigens