Evaluation of T and B lymphocyte function in clinical practice using a flow cytometry based proliferation assay |
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| Authors: | Per Marits,Ann-Charlotte Wikströ m,Dusan Popadic,Ola Winqvist,Sarah Thunberg |
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| Affiliation: | 1. Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden;2. Dept. of Medicine, Karolinska Institutet, Stockholm, Sweden;3. Dept. of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden;4. Inst. of Microbiology and Immunology, University of Belgrade, Serbia;5. Dept. of Oncology and Pathology, Karolinska Institutet, Stockholm, Sweden |
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| Abstract: | The golden standard for functional evaluation of immunodeficiencies is the incorporation of [3H]-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 [3H]-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. |
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| Keywords: | Con A, concanavalin A CPM, counts per minute CV, coefficient of variation FASCIA, flow-cytometric assay for specific cell-mediated immune-response in activated whole blood MMF, mycophenolate mofetil PBMC, peripheral blood mononuclear cells PHA, phytohemagglutinin PID, primary immunodeficiency PPD, purified protein derivative PWM, pokeweed mitogen SEA and SEB, staphylococcal enterotoxin A and B SCID, severe combined immunodeficiency |
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