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In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

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Extracellular Vesicle-associated GARP/TGFβ:LAP Mediates "Infectious" Allo-tolerance.

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|May 30, 2023

Summary

This summary is machine-generated.

Regulatory T cells release exosomes carrying latent transforming growth factor beta (TGFβ):latency-associated peptide (LAP)/glycoprotein A repetitions predominant (GARP) complexes. These exosomal complexes suppress immune responses via direct action or after uptake by bystander T cells.

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Isolation of Murine Lymph Node Stromal Cells

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Published on: August 19, 2014

Area of Science:

Immunology
Cell Biology
Extracellular Vesicles

Background:

Regulatory T cells (Tregs) are crucial for maintaining immune tolerance.
Small extracellular vesicles (sEVs), including exosomes, are key mediators of intercellular communication.
Latent transforming growth factor beta (TGFβ):latency-associated peptide (LAP)/glycoprotein A repetitions predominant (GARP) complexes are known to be immunosuppressive.

Purpose of the Study:

To investigate if TGFβ:LAP/GARP complexes are associated with lymphocyte-derived sEVs in allo-tolerized mice.
To determine if these sEV-associated complexes can suppress immune responses.
To explore the mechanism of TGFβ activation and suppression mediated by these sEVs.

Main Methods:

Induction of allo-tolerance in C57BL/6 mice.
Isolation and restimulation of lymphocytes to induce sEV production.
Ultracentrifugation to isolate sEVs from culture supernatants.
Enzyme-linked immunosorbent assay (ELISA) to detect TGFβ:LAP, GARP, and tetraspanins (CD81, CD63, CD9).
Assessment of TGFβ-dependent immunosuppressive function using a delayed-type hypersensitivity assay.

Main Results:

Lymphocytes from tolerized mice secreted GARP/TGFβ:LAP-coated extracellular vesicles.
GARP/TGFβ:LAP was primarily associated with CD81+ exosomes, similar to IL35.
sEV-bound GARP/TGFβ:LAP demonstrated immunosuppressive activity in both primary and secondary responses.
Secondary suppression required sEV uptake by bystander T cells and reexpression on their surface.

Conclusions:

Exosomal GARP/TGFβ:LAP produced by allo-specific Tregs can mediate immune suppression.
Activation occurs either immediately (primary suppression) or after internalization and reexpression by naive T cells (secondary suppression).
This implicates exosomal TGFβ:LAP and Treg-derived GARP in the network of infectious tolerance.