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Updated: Mar 14, 2026
Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry
Published on: December 16, 2022
Calcium Influx Characteristics During T Lymphocyte Activation Measured with Flow Cytometry
Enikő Birő1, Barna Vásárhelyi2, Gergely Toldi1
1First Department of Pediatrics, Semmelweis University , Budapest H-1083, Hungary.
Insights
This study introduces a novel flow cytometry method for simultaneous calcium influx measurement in T lymphocytes, revealing common patterns in autoimmune diseases like multiple sclerosis and rheumatoid arthritis.
Area of Science:
- Immunology
- Cellular Biology
- Biochemistry
Background:
- T lymphocytes play a crucial role in immune responses and inflammation.
- Traditional single-cell techniques limited simultaneous calcium influx analysis during T lymphocyte activation.
Purpose of the Study:
- To present a novel flow cytometry approach for simultaneous calcium influx recording in multiple lymphocyte subsets.
- To introduce the FacsKin algorithm for robust data analysis of calcium influx.
- To summarize findings on calcium influx characteristics in autoimmune disorders and physiological conditions.
Main Methods:
- Development of a novel flow cytometry technique for simultaneous calcium influx measurement.
- Application of the FacsKin algorithm for quantitative analysis of fluorescent marker data.
- Investigation of calcium influx in multiple sclerosis, rheumatoid arthritis, type 1 diabetes, ankylosing spondylitis, and preeclampsia.
Main Results:
- Identification of dominant calcium influx features common across several investigated autoimmune diseases.
- Characterization of calcium influx patterns under various physiological conditions, including neonatal and pregnancy states.
- Demonstration of the utility of the novel method and algorithm in immunological research.
Conclusions:
- The developed flow cytometry method and FacsKin algorithm enable simultaneous, quantitative assessment of T lymphocyte calcium influx.
- Common calcium influx characteristics are present in multiple autoimmune disorders, suggesting sharedpathogenic mechanisms.
- This approach provides valuable insights into T lymphocyte function in health and disease.
Abstract:
T lymphocytes are of paramount importance in many intercellular reactions, such as the regulation of the inflammatory response and immune reactivity. Until the recent past, single-cell techniques were used for the investigation of calcium influx during T lymphocyte activation. Therefore, over the recent years we have created a novel approach that allows simultaneous recording of calcium influx in several lymphocyte subsets using flow cytometry. Our research group developed a robust algorithm (FacsKin) for the evaluation of the acquired data that fits functions to median values of the fluorescent marker of interest and calculates relevant parameters describing each function. Over the recent years, we have investigated calcium influx characteristics applying this method in a number of autoimmune disorders and under different physiological conditions (such as the neonatal period and pregnancy). In this review, we aim to give a brief summary of our findings and of the common characteristics of calcium influx in the investigated disorders, namely: multiple sclerosis (MS), rheumatoid arthritis (RA), type 1 diabetes mellitus (T1DM), ankylosing spondylitis (AS), and preeclampsia (PE). Based on our results, a number of dominant features were identified that were present in most of the investigated autoimmune diseases.
