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Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus

BACKGROUND: In systemic lupus erythematosus (SLE), autoreactive B cells are thought to develop by-passing immune checkpoints and contribute to its pathogenesis. Toll-like receptor (TLR) 7 and 9 signaling have been implicated in their development and differentiation. Although some B cell subpopulatio...

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Autores principales: Yasaka, Ken, Yamazaki, Tomohide, Sato, Hiroko, Shirai, Tsuyoshi, Cho, Minkwon, Ishida, Koji, Ito, Koyu, Tanaka, Tetsuhiro, Ogasawara, Kouetsu, Harigae, Hideo, Ishii, Tomonori, Fujii, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10577954/
https://www.ncbi.nlm.nih.gov/pubmed/37840148
http://dx.doi.org/10.1186/s13075-023-03186-5
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author Yasaka, Ken
Yamazaki, Tomohide
Sato, Hiroko
Shirai, Tsuyoshi
Cho, Minkwon
Ishida, Koji
Ito, Koyu
Tanaka, Tetsuhiro
Ogasawara, Kouetsu
Harigae, Hideo
Ishii, Tomonori
Fujii, Hiroshi
author_facet Yasaka, Ken
Yamazaki, Tomohide
Sato, Hiroko
Shirai, Tsuyoshi
Cho, Minkwon
Ishida, Koji
Ito, Koyu
Tanaka, Tetsuhiro
Ogasawara, Kouetsu
Harigae, Hideo
Ishii, Tomonori
Fujii, Hiroshi
author_sort Yasaka, Ken
collection PubMed
description BACKGROUND: In systemic lupus erythematosus (SLE), autoreactive B cells are thought to develop by-passing immune checkpoints and contribute to its pathogenesis. Toll-like receptor (TLR) 7 and 9 signaling have been implicated in their development and differentiation. Although some B cell subpopulations such as T-bet + double negative 2 (DN2) cells have been identified as autoreactive in the past few years, because the upregulated surface markers of those cells are not exclusive to them, it is still challenging to specifically target autoreactive B cells in SLE patients. METHODS: Our preliminary expression analysis revealed that phospholipase D4 (PLD4) is exclusively expressed in plasmacytoid dendritic cells (pDCs) and B cells in peripheral blood mononuclear cells (PBMCs) samples. Monoclonal antibodies against human PLD4 were generated, and flow cytometry analyses were conducted for PBMCs from 23 healthy donors (HDs) and 40 patients with SLE. In vitro cell culture was also performed to study the conditions that induce PLD4 in B cells from HDs. Finally, recombinant antibodies were synthesized from subpopulations of PLD4 + B cells from a patient with SLE, and their antinuclear activity was measured through enzyme-linked immunosorbent assay. RESULTS: pDCs from both groups showed comparable frequency of surface PLD4 expression. PLD4 + B cells accounted for only a few percent of HD B cells, whereas they were significantly expanded in patients with SLE (2.1% ± 0.4% vs. 10.8% ± 1.2%, P < 0.005). A subpopulation within PLD4 + B cells whose cell size was comparable to CD38 + CD43 + plasmablasts was defined as “PLD4 + blasts,” and their frequencies were significantly correlated with those of plasmablasts (P < 0.005). PLD4 + blasts phenotypically overlapped with double negative 2 (DN2) cells, and, in line with this, their frequencies were significantly correlated with several clinical markers of SLE. In vitro assay using healthy PBMCs demonstrated that TLR7 or TLR9 stimulation was sufficient to induce PLD4 on the surface of the B cells. Finally, two out of three recombinant antibodies synthesized from PLD4 + blasts showed antinuclear activity. CONCLUSION: PLD4 + B cells, especially “blastic” ones, are likely autoreactive B cells undergoing TLR stimulation. Therefore, PLD4 is a promising target marker in SLE treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13075-023-03186-5.
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spelling pubmed-105779542023-10-17 Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus Yasaka, Ken Yamazaki, Tomohide Sato, Hiroko Shirai, Tsuyoshi Cho, Minkwon Ishida, Koji Ito, Koyu Tanaka, Tetsuhiro Ogasawara, Kouetsu Harigae, Hideo Ishii, Tomonori Fujii, Hiroshi Arthritis Res Ther Research BACKGROUND: In systemic lupus erythematosus (SLE), autoreactive B cells are thought to develop by-passing immune checkpoints and contribute to its pathogenesis. Toll-like receptor (TLR) 7 and 9 signaling have been implicated in their development and differentiation. Although some B cell subpopulations such as T-bet + double negative 2 (DN2) cells have been identified as autoreactive in the past few years, because the upregulated surface markers of those cells are not exclusive to them, it is still challenging to specifically target autoreactive B cells in SLE patients. METHODS: Our preliminary expression analysis revealed that phospholipase D4 (PLD4) is exclusively expressed in plasmacytoid dendritic cells (pDCs) and B cells in peripheral blood mononuclear cells (PBMCs) samples. Monoclonal antibodies against human PLD4 were generated, and flow cytometry analyses were conducted for PBMCs from 23 healthy donors (HDs) and 40 patients with SLE. In vitro cell culture was also performed to study the conditions that induce PLD4 in B cells from HDs. Finally, recombinant antibodies were synthesized from subpopulations of PLD4 + B cells from a patient with SLE, and their antinuclear activity was measured through enzyme-linked immunosorbent assay. RESULTS: pDCs from both groups showed comparable frequency of surface PLD4 expression. PLD4 + B cells accounted for only a few percent of HD B cells, whereas they were significantly expanded in patients with SLE (2.1% ± 0.4% vs. 10.8% ± 1.2%, P < 0.005). A subpopulation within PLD4 + B cells whose cell size was comparable to CD38 + CD43 + plasmablasts was defined as “PLD4 + blasts,” and their frequencies were significantly correlated with those of plasmablasts (P < 0.005). PLD4 + blasts phenotypically overlapped with double negative 2 (DN2) cells, and, in line with this, their frequencies were significantly correlated with several clinical markers of SLE. In vitro assay using healthy PBMCs demonstrated that TLR7 or TLR9 stimulation was sufficient to induce PLD4 on the surface of the B cells. Finally, two out of three recombinant antibodies synthesized from PLD4 + blasts showed antinuclear activity. CONCLUSION: PLD4 + B cells, especially “blastic” ones, are likely autoreactive B cells undergoing TLR stimulation. Therefore, PLD4 is a promising target marker in SLE treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13075-023-03186-5. BioMed Central 2023-10-16 2023 /pmc/articles/PMC10577954/ /pubmed/37840148 http://dx.doi.org/10.1186/s13075-023-03186-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Yasaka, Ken
Yamazaki, Tomohide
Sato, Hiroko
Shirai, Tsuyoshi
Cho, Minkwon
Ishida, Koji
Ito, Koyu
Tanaka, Tetsuhiro
Ogasawara, Kouetsu
Harigae, Hideo
Ishii, Tomonori
Fujii, Hiroshi
Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title_full Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title_fullStr Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title_full_unstemmed Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title_short Phospholipase D4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic T-bet + B cells in systemic lupus erythematosus
title_sort phospholipase d4 as a signature of toll-like receptor 7 or 9 signaling is expressed on blastic t-bet + b cells in systemic lupus erythematosus
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10577954/
https://www.ncbi.nlm.nih.gov/pubmed/37840148
http://dx.doi.org/10.1186/s13075-023-03186-5
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