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Updated: Aug 10, 2026
Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
Modulation of [Ca2+]i in freshly isolated mouse lymphocytes with in vivo priming
Insights
This study shows that calcium mobilization in lymphocytes increases after antigen exposure, correlating with immune response kinetics. This enhanced calcium signaling in sensitized lymphocytes may indicate heightened cellular activity.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Lymphocyte activation involves complex signaling pathways, including intracellular calcium ([Ca2+]i) mobilization.
- Understanding calcium dynamics is crucial for deciphering immune responses to specific antigens.
Purpose of the Study:
- To investigate the pattern of intracellular calcium ([Ca2+]i) mobilization in mouse lymphocytes following immunization with a T-dependent antigen.
- To correlate calcium mobilization with lymphocyte behavior and immunophenotype changes after antigen challenge.
Main Methods:
- In vivo sensitization of mouse lymphocytes with Keyhole Limpet Hemocyanin (KLH).
- In vitro challenge of lymphocytes with Concanavalin A (Con A), Phytohemagglutinin (PHA), or anti-CD3epsilon monoclonal antibody (mAb).
- Measurement of intracellular calcium ([Ca2+]i) levels and analysis of immunophenotypes.
Main Results:
- Baseline [Ca2+]i levels were significantly increased in lymphocytes from immunized mice.
- Calcium mobilization in response to receptor-dependent stimuli (anti-CD3epsilon, PHA, ConA) was enhanced in KLH-primed lymphocytes.
- Increased calcium mobilization correlated with changes in lymphocyte immunophenotype, including increased Ia(k+) B-cells and gammadeltaTCR+ T-cells.
Conclusions:
- Intracellular calcium ([Ca2+]i) mobilization patterns correlate with lymphocyte behavior and the kinetics of the immune response.
- Enhanced calcium signaling in sensitized lymphocytes is linked to an increased number of specific B-cells and gammadeltaTCR+ T-cells.
- Quantitative measurement of [Ca2+]i serves as a valuable biochemical parameter for studying cellular reactions to specific antigens.
Abstract:
Studied at the level of the individual cell, the pattern of [Ca2+]i mobilization of in vivo sensitized mouse lymphocytes by T-dependent antigen (KLH), challenged in vitro by Con A, PHA or anti-CD3epsilon mAb in different periods after immunization, was as follows. In the entire DLN lymphocyte population and in tested T cell subsets from immunized mice, baseline [Ca2+]i was significantly increased and cells were able to respond additionally to stimuli. In KLH-primed DLN lymphocytes, calcium mobilization in response to membrane receptor-dependent stimuli (anti-CD3epsilon, PHA, and ConA) was increased. Enhancement of Ca2+ mobilization is parallel with changed immunophenotype. These findings suggested that: (a) [Ca2+]i mobilization could correlate with lymphocyte behaviour during immunization and that mobilization clearly depended on kinetics of immune reaction; (b) the higher level of activity among sensitized lymphocytes was due to the increased number of specific B-cells (Ia(k+)) and gammadeltaTCR+ cells; (c) the quantitative measurement of [Ca2+]i could be an important biochemical parameter to study cellular reaction to a specific antigen.
