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Thymic stromal lymphopoietin production in DN32.D3 invariant natural killer T (iNKT) cell line and primary mouse liver iNKT cells

Thymic stromal lymphopoietin production in DN32.D3 invariant natural killer T (iNKT) cell line and primary mouse liver iNKT cells
Thymic stromal lymphopoietin production in DN32.D3 invariant natural killer T (iNKT) cell line and primary mouse liver iNKT cells

Background: invariant natural killer T (iNKT) cells are known as the fast responder in allergic inflammation and the source of interleukin (IL)-4, IL-13, and interferon-gamma. Absence of iNKT cells down-regulated thymic stromal lymphopoietin (TSLP) production at the early stage of type 2 immune responses in the airway. However, it has not been reported whether iNKT cells are able to produce TSLP via stimulation of T-cell receptor (TCR).

Objective: we aimed to evaluate TSLP production from iNKT cells by TCR specific stimulations with anti-CD3/CD28 antibodies and α-galactoceramide (α-GalCer).

Methods: DN32.D3 iNKT cell line was stimulated with anti-CD3/CD28 antibodies, and TSLP production was measured in culture supernatants. Next, to confirm the TSLP production in primary mouse iNKT cells, the cells were sorted using α-GalCer-CD1d tetramer from mouse liver, and stimulated with anti-CD3/CD28 antibodies and α-GalCer. Then, cytokine productions were evaluated by enzyme-linked immunosorbent assay and quantitative polymerase chain reaction.

Results: TCR specific stimulation in DN32.D3 cells induced TSLP production as well as signature cytokines of iNKT cells. On the other hand, isolated primary mouse iNKT cells from liver did not show any induction of TSLP by TCR specific stimulations including anti-CD3/CD28 antibodies and α-GalCer, on the contrary to other cytokines.

Conclusion: this study suggested the possibility of TSLP production in iNKT cells, especially from DN32.D3 although primary mouse liver iNKT cells showed a different result.

2233-8276
e10
Choi, Jun-Pyo
93469659-3d0e-42fd-9457-60c614768dbb
Woo, Yeon Duk
e8e10f13-d47a-4b89-bae5-a71907bda3fc
Losol, Purevsuren
13912f45-98d6-4aba-b057-0027993de4c7
Kim, Sae-Hoon
81a4a809-ddb7-4291-897f-62f5177a49ac
Chang, Yoon-Seok
6c517880-794d-4b83-926d-1940310e2ce5
Choi, Jun-Pyo
93469659-3d0e-42fd-9457-60c614768dbb
Woo, Yeon Duk
e8e10f13-d47a-4b89-bae5-a71907bda3fc
Losol, Purevsuren
13912f45-98d6-4aba-b057-0027993de4c7
Kim, Sae-Hoon
81a4a809-ddb7-4291-897f-62f5177a49ac
Chang, Yoon-Seok
6c517880-794d-4b83-926d-1940310e2ce5

Choi, Jun-Pyo, Woo, Yeon Duk, Losol, Purevsuren, Kim, Sae-Hoon and Chang, Yoon-Seok (2021) Thymic stromal lymphopoietin production in DN32.D3 invariant natural killer T (iNKT) cell line and primary mouse liver iNKT cells. Asia Pacific Allergy, 11 (1), e10. (doi:10.5415/apallergy.2021.11.e10).

Record type: Article

Abstract

Background: invariant natural killer T (iNKT) cells are known as the fast responder in allergic inflammation and the source of interleukin (IL)-4, IL-13, and interferon-gamma. Absence of iNKT cells down-regulated thymic stromal lymphopoietin (TSLP) production at the early stage of type 2 immune responses in the airway. However, it has not been reported whether iNKT cells are able to produce TSLP via stimulation of T-cell receptor (TCR).

Objective: we aimed to evaluate TSLP production from iNKT cells by TCR specific stimulations with anti-CD3/CD28 antibodies and α-galactoceramide (α-GalCer).

Methods: DN32.D3 iNKT cell line was stimulated with anti-CD3/CD28 antibodies, and TSLP production was measured in culture supernatants. Next, to confirm the TSLP production in primary mouse iNKT cells, the cells were sorted using α-GalCer-CD1d tetramer from mouse liver, and stimulated with anti-CD3/CD28 antibodies and α-GalCer. Then, cytokine productions were evaluated by enzyme-linked immunosorbent assay and quantitative polymerase chain reaction.

Results: TCR specific stimulation in DN32.D3 cells induced TSLP production as well as signature cytokines of iNKT cells. On the other hand, isolated primary mouse iNKT cells from liver did not show any induction of TSLP by TCR specific stimulations including anti-CD3/CD28 antibodies and α-GalCer, on the contrary to other cytokines.

Conclusion: this study suggested the possibility of TSLP production in iNKT cells, especially from DN32.D3 although primary mouse liver iNKT cells showed a different result.

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More information

Accepted/In Press date: 24 January 2021
e-pub ahead of print date: 28 January 2021
Published date: January 2021

Identifiers

Local EPrints ID: 507152
URI: http://eprints.soton.ac.uk/id/eprint/507152
ISSN: 2233-8276
PURE UUID: fc8e34c1-1922-45b8-acbe-355b6b4cd5fc
ORCID for Purevsuren Losol: ORCID iD orcid.org/0000-0001-7620-1077

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Date deposited: 28 Nov 2025 17:31
Last modified: 29 Nov 2025 03:07

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Contributors

Author: Jun-Pyo Choi
Author: Yeon Duk Woo
Author: Purevsuren Losol ORCID iD
Author: Sae-Hoon Kim
Author: Yoon-Seok Chang

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