Related Experiment Video
Updated: Apr 28, 2026
Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Published on: December 27, 2016
Evaluation of T and B lymphocyte function in clinical practice using a flow cytometry based proliferation assay
Per Marits1, Ann-Charlotte Wikström2, Dusan Popadic3
1Clinical immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Dept. of Medicine, Karolinska Institutet, Stockholm, Sweden.
Insights
The Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) offers a non-radioactive method for assessing lymphocyte function. This assay is a sensitive and rapid alternative to traditional methods for evaluating immunodeficiencies.
Area of Science:
- Immunology
- Cellular Immunology
- Clinical Immunology
Background:
- The gold standard for assessing immunodeficiencies involves [(3)H]-thymidine incorporation, a radioactive method.
- Emerging whole blood proliferation assays offer non-radioactive alternatives with parallel lymphocyte lineage analysis.
- Evaluating novel functional assays is crucial for accurate immunodeficiency diagnosis.
Purpose of the Study:
- To evaluate the Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) as a non-radioactive functional assay.
- To compare FASCIA with the established [(3)H]-thymidine incorporation method.
- To establish reference values and determine optimal conditions for FASCIA.
Main Methods:
- FASCIA was compared against [(3)H]-thymidine incorporation in four severe combined immunodeficiency patients.
- Lymphocyte thresholds for reliable FASCIA responses were determined.
- Reference values were established from 100 healthy donors using mitogenic and antigen-specific stimuli.
- Responses to PWM and SEA+SEB were assessed with immunomodulatory compounds.
Main Results:
- FASCIA demonstrated a reliable assessment of lymphocyte reactivity.
- The assay requires a minimal number of lymphocytes for accurate results.
- Reference values were established for various stimuli in healthy individuals.
- FASCIA showed sensitivity in detecting responses to mitogens and antigens.
Conclusions:
- FASCIA is a rapid, stable, and sensitive functional whole blood assay.
- The assay requires minimal blood volume, making it practical for clinical use.
- FASCIA provides a reliable method for assessing lymphocyte reactivity in patients with immunodeficiencies.
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.
