Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Medicine, Karolinska Institutet, Stockholm, Sweden.
Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden.
Clin Immunol. 2014 Aug;153(2):332-42. doi: 10.1016/j.clim.2014.05.010. Epub 2014 Jun 6.
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.
评估免疫缺陷功能的金标准是使用有丝分裂原刺激进行 [(3)H]-胸苷掺入的增殖测定。最近开发的全血增殖测定法具有平行淋巴细胞谱系分析的优势，并且提供了一种非放射性替代方法。在这里，我们将流式细胞术评估激活全血中的特异性细胞介导免疫反应（FASCIA）与 [(3)H]-胸苷掺入在四名严重联合免疫缺陷患者中的比较。与用有丝分裂原以及抗原特异性刺激物刺激的 100 名健康供体的参考值一起，确定了 FASCIA 中进行可靠反应所需的最少淋巴细胞数的阈值。最后，针对 PWM 和 SEA+SEB 刺激物进行了临床相关免疫调节化合物的反应。我们得出结论，FASCIA 是一种快速、稳定和敏感的全血功能测定法，需要少量全血，可用于可靠评估患者的淋巴细胞反应性。
