title: Evaluation of T and B lymphocyte function in clinical practice using a flow cytometry based proliferation assay.
abstract: The golden standard for functional evaluation of immunodeficiencies is the incorporation of [(3)H]-thymidine in a proliferation assay stimulated with mitogens. Recently developed whole blood proliferation assays have the advantage of parallel lymphocyte lineage analysis and in addition provide a non-radioactive alternative. Here we evaluate the Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) in a comparison with [(3)H]-thymidine incorporation in four patients with severe combined immunodeficiency. The threshold for the minimum number of lymphocytes required for reliable responses in FASCIA is determined together with reference values from 100 healthy donors when stimulated with mitogens as well as antigen specific stimuli. Finally, responses against PWM and SEA+SEB stimuli are conducted with clinically relevant immunomodulatory compounds. We conclude that FASCIA is a rapid, stable and sensitive functional whole blood assay that requires small amounts of whole blood that can be used for reliable assessment of lymphocyte reactivity in patients.
authors.affiliation: Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Per
authors.initials: P
authors.lastname: Marits
authors.affiliation: Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Ann-Charlotte
authors.initials: AC
authors.lastname: Wikström
authors.affiliation: Inst. of Microbiology and Immunology, University of Belgrade, Serbia.
authors.forename: Dusan
authors.initials: D
authors.lastname: Popadic
authors.affiliation: Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Ola
authors.initials: O
authors.lastname: Winqvist
authors.affiliation: Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Oncology and Pathology, Karolinska Institutet, Stockholm, Sweden. Electronic address: sara.thunberg@karolinska.se.
authors.forename: Sarah
authors.initials: S
authors.lastname: Thunberg
mesh_terms: D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D049109:Cell Proliferation; D003907:Dexamethasone / Q000276:immunology / Q000494:pharmacology; D004768:Enterotoxins / Q000276:immunology / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007166:Immunosuppressive Agents / Q000276:immunology / Q000494:pharmacology; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D020381:Interleukin-17 / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D018655:Lymphocyte Count; D011043:Pokeweed Mitogens / Q000276:immunology / Q000494:pharmacology; D015203:Reproducibility of Results; D016511:Severe Combined Immunodeficiency / Q000175:diagnosis / Q000276:immunology*; D020123:Sirolimus / Q000276:immunology / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D016559:Tacrolimus / Q000276:immunology / Q000494:pharmacology
chemical_list: D004768:Enterotoxins; D007166:Immunosuppressive Agents; D020381:Interleukin-17; D011043:Pokeweed Mitogens; C026066:enterotoxin A, Staphylococcal; C031818:enterotoxin B, staphylococcal; D003907:Dexamethasone; D007371:Interferon-gamma; D020123:Sirolimus; D016559:Tacrolimus
keywords: Flow cytometry; Lymphocyte proliferation assay; Primary immune deficiency