Journal of Molecular Histology

, Volume 50, Issue 4, pp 325–333 | Cite as

In vitro assessment of PD-L1+ microvesicles in the cyst fluid of non-syndromic odontogenic keratocysts

  • Qi-Wen Man
  • Wen-Qun Zhong
  • Yi-Fang Zhao
  • Bing Liu
  • Yi ZhaoEmail author
Original Paper


Odontogenic keratocysts (OKCs) are jaw cystic lesions which are characterized by local invasion and high recurrence rate. The majority of OKCs are exposed to microorganisms and occur along with focal inflammatory infiltrates. Cyst fluids are biological fluids that contain a large content of cytokines and immune globulins. Inhibitory receptor such as programmed death receptor 1 (PD-1) and its ligand programmed death-ligand 1 (PD-L1), which can induce a coinhibitory signal in activated T cells, plays a vital role in the differentiation, exhaustion and apoptosis of T cells. Cell derived microvesicles, carrying a cargo of functional proteins, nucleic acids and lipids, are important communication tools in the development of diseases. However, the expression of PD-L1 in OKCs tissues and whether PD-L1 could be carried by microvesicles are unexplored. Presently, we have isolated cyst fluid microvesicles and identified cell derived PD-L1+ cyst fluid microvesicles. PD-L1 was located in the membrane of the cyst fluid microvesicles. The main cellular origins of PD-L1+ cyst fluid microvesicles were dendritic cells followed by lymphocytes. Elevated PD-L1+ cyst fluid microvesicles were detected in the OKCs compared with dentigerous cysts. Isolated cyst fluid microvesicles could bind to the membrane of activated CD8 T cells and inhibit proliferation of stimulated peripheral blood CD8 T cells. In conclusion, the present study suggests that elevated PD-L1+ cyst fluid microvesicles might be related with the cyst development of OKCs.


Odontogenic keratocysts PD-L1 Microvesicles Cyst fluids 



Odontogenic keratocysts


Dentigerous cysts




Cyst fluid microvesicles


Carboxyfluoresceinsuccinimidyl ester



The authors are grateful to all the pathology doctors from Hospital of Stomatology Wuhan University and technician Min Juan from the Centre Institute of Virology, Chinese Academy of Science for supporting our flow cytometric analysis.


This study was supported by Grants National Natural Science Foundation of China to Dr. Bing Liu (No. 81872203) and Dr. Wen-Qun Zhong (No. 81800994).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interests.


  1. Ahlfors E, Larsson A, Sjogren S (1984) The odontogenic keratocyst: a benign cystic tumor? J Oral Maxillofac Surg 42:10–19CrossRefGoogle Scholar
  2. Al-Moraissi EA et al (2017) What surgical treatment has the lowest recurrence rate following the management of keratocystic odontogenic tumor? A large systematic review and meta-analysis. J Craniomaxillofac Surg 45:131–144CrossRefGoogle Scholar
  3. Aragaki T et al (2010) Comprehensive keratin profiling reveals different histopathogenesis of keratocystic odontogenic tumor and orthokeratinized odontogenic cyst. Hum Pathol 41:1718–1725CrossRefGoogle Scholar
  4. Barnes L, Eveson JW, Reichart P, Sidransky D (2005) Pathology and genetics of head and neck tumours, 3rd edn. WHO Classification of Tumours, Geneva, p 306Google Scholar
  5. Baumeister SH, Freeman GJ, Dranoff G, Sharpe AH (2016) Coinhibitory pathways in immunotherapy for cancer. Annu Rev Immunol 34:539–573CrossRefGoogle Scholar
  6. Browne RM (1976) Some observations on the fluids of odontogenic cysts. J Oral Pathol 5:74–87CrossRefGoogle Scholar
  7. Cha YH et al (2017) Frequent oncogenic BRAF V600E mutation in odontogenic keratocyst. Oral Oncol 74:62–67CrossRefGoogle Scholar
  8. Chen G et al (2018) Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response. Nature 560:382–386CrossRefGoogle Scholar
  9. Diniz MG, Gomes CC, de Sousa SF, Xavier GM, Gomez RS (2017) Oncogenic signalling pathways in benign odontogenic cysts and tumours. Oral Oncol 72:165–173CrossRefGoogle Scholar
  10. El-Naggar AK, Chan JKC, Grandis JR, Takata T, Slootweg PJ (2005) WHO Classification of Head and Neck Tumours, 4th edn. WHO Classification of Tumours, Geneva, p 235Google Scholar
  11. Gong J, Jaiswal R, Dalla P, Luk F, Bebawy M (2015) Microparticles in cancer: A review of recent developments and the potential for clinical application. Semin Cell Dev Biol 40:35–40CrossRefGoogle Scholar
  12. Grisendi G et al (2015) Detection of microparticles from human red blood cells by multiparametric flow cytometry. Blood Transfus 13:274–280Google Scholar
  13. Hayashi M et al (2008) Profiling of radicular cyst and odontogenic keratocyst cytokine production suggests common growth mechanisms. J Endod 34:14–21CrossRefGoogle Scholar
  14. Hazarika M et al (2017) U.S. FDA Approval Summary: nivolumab for treatment of unresectable or metastatic melanoma following progression on Ipilimumab. Clin Cancer Res 23:3484–3488CrossRefGoogle Scholar
  15. Kaminagakura E et al (2013) Keratocyst of the buccal mucosa: case report and immunohistochemical comparative study with sporadic intraosseous keratocystic odontogenic tumor. Oral Surg Oral Med Oral Pathol Oral Radiol 116:e387–392CrossRefGoogle Scholar
  16. Kichi E, Enokiya Y, Muramatsu T, Hashimoto S, Inoue T, Abiko Y, Shimono M (2005) Cell proliferation, apoptosis and apoptosis-related factors in odontogenic keratocysts and in dentigerous cysts. J Oral Pathol Med 34:280–286CrossRefGoogle Scholar
  17. Koirala P et al (2016) Immune infiltration and PD-L1 expression in the tumor microenvironment are prognostic in osteosarcoma. Sci Rep 6:30093CrossRefGoogle Scholar
  18. Kolár Z, Geierová M, Bouchal J, Pazdera J, Zboril V, Tvrdý P (2006) Immunohistochemical analysis of the biological potential of odontogenic keratocysts. J Oral Pathol Med 35:75–80CrossRefGoogle Scholar
  19. Larkins E et al (2017) FDA Approval Summary: pembrolizumab for the treatment of recurrent or metastatic head and neck squamous cell carcinoma with disease progression on or after platinum-containing chemotherapy. Oncologist 22:873–878CrossRefGoogle Scholar
  20. Leung YY, Lau SL, Tsoi KY, Ma HL, Ng CL (2016) Results of the treatment of keratocystic odontogenic tumours using enucleation and treatment of the residual bony defect with Carnoy’s solution. Int J Oral Maxillofac Surg 45:1154–1158CrossRefGoogle Scholar
  21. Li TJ (2011) The odontogenic keratocyst: a cyst, or a cystic neoplasm? J Dent Res 90:133–142CrossRefGoogle Scholar
  22. Li CW et al (2016) Glycosylation and stabilization of programmed death ligand-1 suppresses T-cell activity. Nat Commun 7:12632CrossRefGoogle Scholar
  23. Man QW, Zhang LZ, Zhao Y, Liu JY, Zheng YY, Zhao YF, Liu B (2018a) Lymphocytederived microparticles stimulate osteoclastogenesis by inducing RANKL in fibroblasts of odontogenic keratocysts. Oncol Rep 40:3335–3345Google Scholar
  24. Man QW et al (2018b) Increased level of cell-derived microparticles in the cyst fluids of odontogenic keratocysts. Int J Oncol 52:1863–1874Google Scholar
  25. Matos FT et al (2013) Immunophenotypic characterization and distribution of dendritic cells in odontogenic cystic lesions. Oral Dis 19:85–91CrossRefGoogle Scholar
  26. Mello LA et al (2013) CD1a-positive Langerhans cells and their relationship with E-cadherin in ameloblastomas and keratocystic odontogenic tumors. J Oral Pathol Med 42:454–461CrossRefGoogle Scholar
  27. Muller L, Simms P, Hong CS, Nishimura MI, Jackson EK, Watkins SC, Whiteside TL (2017) Human tumor-derived exosomes (TEX) regulate Treg functions via cell surface signaling rather than uptake mechanisms. Oncoimmunology 6:e1261243CrossRefGoogle Scholar
  28. Nieuwland R et al (2000) Cellular origin and procoagulant properties of microparticles in meningococcal sepsis. Blood 95:930–935Google Scholar
  29. Pavelić B, Levanat S, Crnić I, Kobler P, Anić I, Manojlović S, Sutalo J (2001) PTCH gene altered in dentigerous cysts. J Oral Pathol Med 30:569–576CrossRefGoogle Scholar
  30. Piattelli A, Rubini C, Iezzi G, Fioroni M (2002) CD1a-positive cells in odontogenic cysts. J Endod 28:267–268CrossRefGoogle Scholar
  31. Piccin A, Murphy WG, Smith OP (2007) Circulating microparticles: pathophysiology and clinical implications. Blood Rev 21:157–171CrossRefGoogle Scholar
  32. Pogrel MA, Jordan RC (2004) Marsupialization as a definitive treatment for the odontogenic keratocyst. J Oral Maxillofac Surg 62:651–655CrossRefGoogle Scholar
  33. Press JZ et al (2012) Microparticles from ovarian carcinomas are shed into ascites and promote cell migration. Int J Gynecol Cancer 22:546–552CrossRefGoogle Scholar
  34. Ren JG et al (2016a) Elevated level of circulating platelet-derived microparticles in oral cancer. J Dent Res 95:87–93CrossRefGoogle Scholar
  35. Ren JG et al (2016b) Clinical significance and roles in angiogenesis of circulating microparticles in oral cancer. J Dent Res 95:860–867CrossRefGoogle Scholar
  36. Schumacher TN, Schreiber RD (2015) Neoantigens in cancer immunotherapy. Science 348:69–74CrossRefGoogle Scholar
  37. Sharif FN, Oliver R, Sweet C, Sharif MO (2015) Interventions for the treatment of keratocystic odontogenic tumours. Cochrane Database Syst Rev 5:CD008464Google Scholar
  38. Shear M (2002) The aggressive nature of the odontogenic keratocyst: is it a benign cystic neoplasm? Part 1. Clinical and early experimental evidence of aggressive behaviour. Oral Oncol 38:219–226CrossRefGoogle Scholar
  39. Shi L, Chen S, Yang L, Li Y (2013) The role of PD-1 and PD-L1 in T-cell immune suppression in patients with hematological malignancies. J Hematol Oncol 6:74CrossRefGoogle Scholar
  40. Skaug N (1973) Proteins in fluid from non-keratinizing jaw cysts. 2. Concentrations of total protein, some protein fractions and nitrogen. J Oral Pathol 2:280–291CrossRefGoogle Scholar
  41. Sul J, Blumenthal GM, Jiang X, He K, Keegan P, Pazdur R (2016) FDA approval summary: pembrolizumab for the treatment of patients With metastatic non-small cell lung cancer whose tumors express programmed death-ligand 1. Oncologist 21:643–650CrossRefGoogle Scholar
  42. Thompson L (2006) World Health Organization classification of tumours: pathology and genetics of head and neck tumours. Ear Nose Throat J 85:74CrossRefGoogle Scholar
  43. Wei SC, Duffy CR, Allison JP (2018) Fundamental mechanisms of immune checkpoint blockade therapy. Cancer Discov 8:1069–1086CrossRefGoogle Scholar
  44. Williams C, Royo F, Aizpurua-Olaizola O (2018) Glycosylation of extracellular vesicles: current knowledge, tools and clinical perspectives. J Extracell Vesicles 7:1442985CrossRefGoogle Scholar
  45. Wright JM, Vered M (2017) Update from the 4th edition of the World Health Organization Classification of head and neck tumours: odontogenic and maxillofacial bone tumors. Head Neck Pathol 11:68–77CrossRefGoogle Scholar
  46. Xu JX et al (2017) FDA Approval Summary: nivolumab in advanced renal cell carcinoma after anti-angiogenic therapy and exploratory predictive biomarker analysis. Oncologist 22:311–317CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Qi-Wen Man
    • 1
  • Wen-Qun Zhong
    • 1
  • Yi-Fang Zhao
    • 1
    • 2
  • Bing Liu
    • 1
    • 2
  • Yi Zhao
    • 1
    • 3
    Email author
  1. 1.State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of StomatologyWuhan UniversityWuhanChina
  2. 2.Department of Oral and Maxillofacial Surgery, Wuhan University School of StomatologyWuhan UniversityWuhanChina
  3. 3.Department of Prosthodontics, School and Hospital of StomatologyWuhan UniversityWuhanChina

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