Article Text

Download PDFPDF

Extended report
Semaphorin 4D inhibits neutrophil activation and is involved in the pathogenesis of neutrophil-mediated autoimmune vasculitis
  1. Masayuki Nishide1,2,3,
  2. Satoshi Nojima2,3,4,
  3. Daisuke Ito2,3,5,
  4. Hyota Takamatsu1,3,2,
  5. Shohei Koyama1,3,2,
  6. Sujin Kang2,3,6,
  7. Tetsuya Kimura1,3,2,
  8. Keiko Morimoto1,3,2,
  9. Takashi Hosokawa1,3,2,
  10. Yoshitomo Hayama1,3,2,
  11. Yuhei Kinehara1,3,2,
  12. Yasuhiro Kato1,3,2,
  13. Takeshi Nakatani1,3,2,
  14. Yoshimitsu Nakanishi1,3,2,
  15. Takeshi Tsuda2,3,7,
  16. Jeong Hoon Park2,
  17. Toru Hirano1,
  18. Yoshihito Shima1,
  19. Masashi Narazaki1,
  20. Eiichi Morii4,
  21. Atsushi Kumanogoh1,3,2
  1. 1 Department of Respiratory Medicine and Clinical Immunology, Osaka University Graduate School of Medicine, Suita City, Osaka, Japan
  2. 2 Laboratory of Immunopathology, World Premier International Immunology Frontier Research Center, Suita City, Osaka, Japan
  3. 3 The Japan Agency for Medical Research and Development–Core Research for Evolutional Science and Technology (AMED–CREST), Japan
  4. 4 Department of Pathology, Osaka University Graduate School of Medicine, Suita City, Osaka, Japan
  5. 5 Department of Nephrology, Osaka University Graduate School of Medicine, Suita City, Osaka, Japan
  6. 6 Department of Clinical Application of Biologics, Osaka University Graduate School of Medicine, Suita City, Osaka, Japan
  7. 7 Department of Otorhinolaryngology–Head and Neck Surgery, Osaka University Graduate School of Medicine, Suita City, Osaka, Japan
  1. Correspondence to Assistant Professor Satoshi Nojima, Department of PathologyOsaka University Graduate School of MedicineSuita City, OsakaJapan; s_nojima{at} and Professor Atsushi Kumanogoh, Department of Respiratory Medicine, Allergy and Rheumatic Diseases, Suita City 565-0871, Osaka, Japan; kumanogo{at}


Objectives Inappropriate activation of neutrophils plays a pathological role in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). The aim of this study was to investigate the functions of semaphorin 4D (SEMA4D) in regulation of neutrophil activation, and its involvement in AAV pathogenesis.

Methods Serum levels of soluble SEMA4D were evaluated by ELISA. Blood cell-surface expression of membrane SEMA4D was evaluated by flow cytometry. To determine the functional interactions between neutrophil membrane SEMA4D and endothelial plexin B2, wild-type and SEMA4D −/− mice neutrophils were cultured with an endothelial cell line (MS1) stained with SYTOX green, and subjected to neutrophil extracellular trap (NET) formation assays. The efficacy of treating human neutrophils with recombinant plexin B2 was assessed by measuring the kinetic oxidative burst and NET formation assays.

Results Serum levels of soluble SEMA4D were elevated in patients with AAV and correlated with disease activity scores. Cell-surface expression of SEMA4D was downregulated in neutrophils from patients with AAV, a consequence of proteolytic cleavage of membrane SEMA4D. Soluble SEMA4D exerted pro-inflammatory effects on endothelial cells. Membranous SEMA4D on neutrophils bound to plexin B2 on endothelial cells, and this interaction decreased NET formation. Recombinant plexin B2 suppressed neutrophil Rac1 activation through SEMA4D’s intracellular domain, and inhibited pathogen-induced or ANCA-induced oxidative burst and NET formation.

Conclusions Neutrophil surface SEMA4D functions as a negative regulator of neutrophil activation. Proteolytic cleavage of SEMA4D as observed in patients with AAV may amplify neutrophil-mediated inflammatory responses. SEMA4D is a promising biomarker and potential therapeutic target for AAV.

  • Autoimmune Diseases
  • Granulomatosis with polyangiitis
  • Systemic vasculitis
  • Inflammation

This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.


  • Contributors AK designed and supervised the project. MN carried out most of experiments. SN, DI, TT and YN conducted some of experiments. HT, SK, SK, TK, KM, EM and AK contributed to preparation of materials and provided advice on project planning and data interpretation. MN, TH, YH, YK, YK, TN, TH, YS and MN recruited and clinically characterised patients. MN, SN, JP and AK wrote the manuscript. SN and AK provided funding for the study. All authors contributed to the discussion of the results and edited and approved the final version.

  • Competing interests None declared.

  • Patient consent All human samples were obtained after informed consent was provided by the subjects, in accordance with the Declaration of Helsinki and with approval from the ethical review board of the Graduate School of Medicine, Osaka University, Japan (no. 10237).

  • Ethics approval The ethical review board of the Graduate School of Medicine, Osaka University, Japan (no. 10237). This study was registered in the University Hospital Medical Information Network Clinical Trials Registry (UMIN000013076).

  • Provenance and peer review Not commissioned; externally peer reviewed.