Soluble RANKL Cleaved from Activated Lymphocytes by TNF-α-Converting Enzyme Contributes to Osteoclastogenesis in Periodontitis

Hiroyuki Kanzaki, Seicho Makihira, Maiko Suzuki, Takenobu Ishii, Alexandru Movila, Josefine Hirschfeld, Hani Mawardi, Xiaoping Lin, Xiaozhe Han, Martin A Taubman, Toshihisa Kawai

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Host immune responses play a key role in promoting bone resorption in periodontitis via receptor activator of NF-κB ligand (RANKL)-dependent osteoclastogenesis. Both membrane-bound RANKL (mRANKL) expressed on lymphocytes and soluble RANKL (sRANKL) are found in periodontal lesions. However, the underlying mechanism and cellular source of sRANKL release and its biological role in periodontitis are unclear. TNF-α-converting enzyme (TACE) is reported to cleave the following: 1) precursor TNF-α with release of mature, soluble TNF-α and 2) mRANKL with release of sRANKL. Both soluble TNF-α and sRANKL are found in the periodontitis lesion, leading to the hypothesis that TACE expressed on lymphocytes is engaged in RANKL shedding and that the resulting sRANKL induces osteoclastogenesis. In the current study, upon stimulating PBLs with mitogens in vitro, RANKL expression, sRANKL secretion, and TACE expression were all upregulated. Among the four putative mRANKL sheddases examined in neutralization assays, TACE was the only functional sheddase able to cleave mRANKL expressed on PBL. Moreover, PBL culture supernatant stimulated with mitogens in the presence of anti-TACE Ab or anti-RANKL Ab showed a marked reduction of osteoclastogenesis from osteoclast precursors, indicating that TACE-mediated sRANKL may possess sufficient osteoclastogenic activity. According to double-color confocal microscopy, B cells expressed a more pronounced level of RANKL and TACE expression than T cells or monocytes in periodontally diseased gingiva. Conditioned medium of patients' gingival lymphocyte culture increased in vitro osteoclastogenic activity, which was suppressed by the addition of anti-TACE Ab and anti-RANKL Ab. Therefore, TACE-mediated cleavage of sRANKL from activated lymphocytes, especially B cells, can promote osteoclastogenesis in periodontitis.

Original languageEnglish (US)
Pages (from-to)3871-3883
Number of pages13
JournalJournal of Immunology
Volume197
Issue number10
DOIs
StatePublished - Nov 15 2016
Externally publishedYes

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Periodontitis
Osteogenesis
Lymphocytes
Membranes
Enzymes
Mitogens
B-Lymphocytes
Gingiva
Osteoclasts
Bone Resorption
Conditioned Culture Medium
Confocal Microscopy
Monocytes
Color
Ligands
T-Lymphocytes
In Vitro Techniques

Keywords

  • ADAM17 Protein/genetics
  • B-Lymphocytes/drug effects
  • Bone Resorption
  • Cells, Cultured
  • Gingiva/cytology
  • Humans
  • Lipopolysaccharides/immunology
  • Lymphocyte Activation
  • Male
  • Monocytes/immunology
  • Osteoclasts/immunology
  • Osteogenesis
  • Periodontitis/immunology
  • RANK Ligand/genetics
  • Solubility
  • T-Lymphocytes/immunology

Cite this

Soluble RANKL Cleaved from Activated Lymphocytes by TNF-α-Converting Enzyme Contributes to Osteoclastogenesis in Periodontitis. / Kanzaki, Hiroyuki; Makihira, Seicho; Suzuki, Maiko; Ishii, Takenobu; Movila, Alexandru; Hirschfeld, Josefine; Mawardi, Hani; Lin, Xiaoping; Han, Xiaozhe; Taubman, Martin A; Kawai, Toshihisa.

In: Journal of Immunology, Vol. 197, No. 10, 15.11.2016, p. 3871-3883.

Research output: Contribution to journalArticle

Kanzaki, H, Makihira, S, Suzuki, M, Ishii, T, Movila, A, Hirschfeld, J, Mawardi, H, Lin, X, Han, X, Taubman, MA & Kawai, T 2016, 'Soluble RANKL Cleaved from Activated Lymphocytes by TNF-α-Converting Enzyme Contributes to Osteoclastogenesis in Periodontitis', Journal of Immunology, vol. 197, no. 10, pp. 3871-3883. https://doi.org/10.4049/jimmunol.1601114
Kanzaki, Hiroyuki ; Makihira, Seicho ; Suzuki, Maiko ; Ishii, Takenobu ; Movila, Alexandru ; Hirschfeld, Josefine ; Mawardi, Hani ; Lin, Xiaoping ; Han, Xiaozhe ; Taubman, Martin A ; Kawai, Toshihisa. / Soluble RANKL Cleaved from Activated Lymphocytes by TNF-α-Converting Enzyme Contributes to Osteoclastogenesis in Periodontitis. In: Journal of Immunology. 2016 ; Vol. 197, No. 10. pp. 3871-3883.
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abstract = "Host immune responses play a key role in promoting bone resorption in periodontitis via receptor activator of NF-κB ligand (RANKL)-dependent osteoclastogenesis. Both membrane-bound RANKL (mRANKL) expressed on lymphocytes and soluble RANKL (sRANKL) are found in periodontal lesions. However, the underlying mechanism and cellular source of sRANKL release and its biological role in periodontitis are unclear. TNF-α-converting enzyme (TACE) is reported to cleave the following: 1) precursor TNF-α with release of mature, soluble TNF-α and 2) mRANKL with release of sRANKL. Both soluble TNF-α and sRANKL are found in the periodontitis lesion, leading to the hypothesis that TACE expressed on lymphocytes is engaged in RANKL shedding and that the resulting sRANKL induces osteoclastogenesis. In the current study, upon stimulating PBLs with mitogens in vitro, RANKL expression, sRANKL secretion, and TACE expression were all upregulated. Among the four putative mRANKL sheddases examined in neutralization assays, TACE was the only functional sheddase able to cleave mRANKL expressed on PBL. Moreover, PBL culture supernatant stimulated with mitogens in the presence of anti-TACE Ab or anti-RANKL Ab showed a marked reduction of osteoclastogenesis from osteoclast precursors, indicating that TACE-mediated sRANKL may possess sufficient osteoclastogenic activity. According to double-color confocal microscopy, B cells expressed a more pronounced level of RANKL and TACE expression than T cells or monocytes in periodontally diseased gingiva. Conditioned medium of patients' gingival lymphocyte culture increased in vitro osteoclastogenic activity, which was suppressed by the addition of anti-TACE Ab and anti-RANKL Ab. Therefore, TACE-mediated cleavage of sRANKL from activated lymphocytes, especially B cells, can promote osteoclastogenesis in periodontitis.",
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T1 - Soluble RANKL Cleaved from Activated Lymphocytes by TNF-α-Converting Enzyme Contributes to Osteoclastogenesis in Periodontitis

AU - Kanzaki, Hiroyuki

AU - Makihira, Seicho

AU - Suzuki, Maiko

AU - Ishii, Takenobu

AU - Movila, Alexandru

AU - Hirschfeld, Josefine

AU - Mawardi, Hani

AU - Lin, Xiaoping

AU - Han, Xiaozhe

AU - Taubman, Martin A

AU - Kawai, Toshihisa

N1 - Copyright © 2016 by The American Association of Immunologists, Inc.

PY - 2016/11/15

Y1 - 2016/11/15

N2 - Host immune responses play a key role in promoting bone resorption in periodontitis via receptor activator of NF-κB ligand (RANKL)-dependent osteoclastogenesis. Both membrane-bound RANKL (mRANKL) expressed on lymphocytes and soluble RANKL (sRANKL) are found in periodontal lesions. However, the underlying mechanism and cellular source of sRANKL release and its biological role in periodontitis are unclear. TNF-α-converting enzyme (TACE) is reported to cleave the following: 1) precursor TNF-α with release of mature, soluble TNF-α and 2) mRANKL with release of sRANKL. Both soluble TNF-α and sRANKL are found in the periodontitis lesion, leading to the hypothesis that TACE expressed on lymphocytes is engaged in RANKL shedding and that the resulting sRANKL induces osteoclastogenesis. In the current study, upon stimulating PBLs with mitogens in vitro, RANKL expression, sRANKL secretion, and TACE expression were all upregulated. Among the four putative mRANKL sheddases examined in neutralization assays, TACE was the only functional sheddase able to cleave mRANKL expressed on PBL. Moreover, PBL culture supernatant stimulated with mitogens in the presence of anti-TACE Ab or anti-RANKL Ab showed a marked reduction of osteoclastogenesis from osteoclast precursors, indicating that TACE-mediated sRANKL may possess sufficient osteoclastogenic activity. According to double-color confocal microscopy, B cells expressed a more pronounced level of RANKL and TACE expression than T cells or monocytes in periodontally diseased gingiva. Conditioned medium of patients' gingival lymphocyte culture increased in vitro osteoclastogenic activity, which was suppressed by the addition of anti-TACE Ab and anti-RANKL Ab. Therefore, TACE-mediated cleavage of sRANKL from activated lymphocytes, especially B cells, can promote osteoclastogenesis in periodontitis.

AB - Host immune responses play a key role in promoting bone resorption in periodontitis via receptor activator of NF-κB ligand (RANKL)-dependent osteoclastogenesis. Both membrane-bound RANKL (mRANKL) expressed on lymphocytes and soluble RANKL (sRANKL) are found in periodontal lesions. However, the underlying mechanism and cellular source of sRANKL release and its biological role in periodontitis are unclear. TNF-α-converting enzyme (TACE) is reported to cleave the following: 1) precursor TNF-α with release of mature, soluble TNF-α and 2) mRANKL with release of sRANKL. Both soluble TNF-α and sRANKL are found in the periodontitis lesion, leading to the hypothesis that TACE expressed on lymphocytes is engaged in RANKL shedding and that the resulting sRANKL induces osteoclastogenesis. In the current study, upon stimulating PBLs with mitogens in vitro, RANKL expression, sRANKL secretion, and TACE expression were all upregulated. Among the four putative mRANKL sheddases examined in neutralization assays, TACE was the only functional sheddase able to cleave mRANKL expressed on PBL. Moreover, PBL culture supernatant stimulated with mitogens in the presence of anti-TACE Ab or anti-RANKL Ab showed a marked reduction of osteoclastogenesis from osteoclast precursors, indicating that TACE-mediated sRANKL may possess sufficient osteoclastogenic activity. According to double-color confocal microscopy, B cells expressed a more pronounced level of RANKL and TACE expression than T cells or monocytes in periodontally diseased gingiva. Conditioned medium of patients' gingival lymphocyte culture increased in vitro osteoclastogenic activity, which was suppressed by the addition of anti-TACE Ab and anti-RANKL Ab. Therefore, TACE-mediated cleavage of sRANKL from activated lymphocytes, especially B cells, can promote osteoclastogenesis in periodontitis.

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KW - B-Lymphocytes/drug effects

KW - Bone Resorption

KW - Cells, Cultured

KW - Gingiva/cytology

KW - Humans

KW - Lipopolysaccharides/immunology

KW - Lymphocyte Activation

KW - Male

KW - Monocytes/immunology

KW - Osteoclasts/immunology

KW - Osteogenesis

KW - Periodontitis/immunology

KW - RANK Ligand/genetics

KW - Solubility

KW - T-Lymphocytes/immunology

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DO - 10.4049/jimmunol.1601114

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VL - 197

SP - 3871

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JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

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