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Updated: Aug 1, 2025
Imaging the Human Immunological Synapse
Published on: December 26, 2019
T Cell Immunological Synaptosomes: Definition and Isolation
Hye-Ran Kim1,2, Jeong-Su Park1, Na-Young Kim1
1School of Life Sciences, Immune Synapse and Cell Therapy Research Center, Gwangju Institute of Science and Technology (GIST), Gwangju, South Korea.
Abstract:
In addition to microvilli's role as structural scaffold for TCR clustering, we recently discovered a novel function as message senders. We found that microvilli are separated from the T cell body shortly upon TCR stimulation and vesiculated to form T cell microvilli particles (TMPs), a new type of membrane vesicles. TMPs and synaptic ectosomes, which bud from the synaptic cleft, constitute "T cell immunological synaptosomes (TISs)" and act as conveyors of T cell messages or traits to cognate antigen-presenting cells. In practice, it is almost impossible to distinguish between TMPs and synaptic ectosomes. Here, we describe a newly developed protocol to isolate TISs from activated T cells using antibody-immobilized agarose beads and density gradient ultracentrifugation. We further describe the methods for TIS quantification with flow cytometry and to evaluate TIS efficacy on dendritic cells.
Insights
Microvilli release T cell microvilli particles (TMPs) upon stimulation, forming T cell immunological synaptosomes (TISs). These TISs transfer messages to antigen-presenting cells, revealing a novel immune communication pathway.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Microvilli traditionally serve as structural scaffolds for T cell receptor (TCR) clustering.
- Recent findings indicate microvilli possess a novel function in intercellular communication.
Purpose of the Study:
- To investigate the role of microvilli in T cell signaling and intercellular communication.
- To describe a method for isolating and analyzing T cell immunological synaptosomes (TISs).
Main Methods:
- Activated T cells were stimulated to induce microvilli vesiculation.
- T cell microvilli particles (TMPs) and synaptic ectosomes were isolated to form TISs using antibody-immobilized agarose beads and density gradient ultracentrifugation.
- Flow cytometry was employed for TIS quantification and assessment of their effects on dendritic cells.
Main Results:
- Microvilli detach upon TCR stimulation, forming T cell microvilli particles (TMPs).
- TMPs and synaptic ectosomes combine to form T cell immunological synaptosomes (TISs).
- TISs function as conveyors of T cell messages to antigen-presenting cells.
Conclusions:
- TISs represent a newly identified mechanism for T cell communication.
- The developed protocol enables the isolation, quantification, and functional evaluation of TISs.
- This discovery sheds light on novel pathways of immune cell interaction and information transfer.
