Analysis of Store-Operated Ca²⁺ Entry in Primary T Cells

Area of Science:

• Immunology
• Cell Biology
• Biochemistry

Background:

• Calcium ions (Ca²⁺) are vital second messengers in cellular signal transduction.
• In T cells, Ca²⁺ signaling via Store-Operated Ca²⁺ Entry (SOCE) is critical for immune responses.
• SOCE involves endoplasmic reticulum (ER) Ca²⁺ store depletion and subsequent extracellular Ca²⁺ influx through Ca²⁺ release-activated Ca²⁺ (CRAC) channels.

Purpose of the Study:

• To review general concepts of Ca²⁺ signaling in T cells.
• To discuss the use of ratiometric Ca²⁺-sensitive dyes for monitoring SOCE in primary murine T cells.
• To present detailed protocols for measuring and quantifying SOCE in T cells.

Main Methods:

• Utilizing ratiometric Ca²⁺-sensitive chemical dyes.
• Employing flow cytometry for SOCE measurement.
• Applying fluorescent microplate reader and single-cell imaging techniques.

Main Results:

• Detailed protocols for measuring SOCE in mouse T cells are provided.
• Advantages and disadvantages of different cytosolic Ca²⁺ dynamics detection methods are discussed.
• Quantification guidelines for SOCE are offered.

Conclusions:

• The presented protocols are adaptable for human T cells and other cell types.
• Accurate measurement of SOCE is essential for understanding T cell function.
• This work facilitates research into T cell-mediated adaptive immunity.