Advertisement

Virchows Archiv

, Volume 474, Issue 2, pp 201–207 | Cite as

Evaluation of tumor-infiltrating lymphocytes in osteosarcomas of the jaws: a multicenter study

  • Pollianna Muniz Alves
  • José Alcides Almeida de Arruda
  • Diego Antônio Costa Arantes
  • Sara Ferreira Santos Costa
  • Lucas Lacerda Souza
  • Hélder Antônio Rebelo Pontes
  • Felipe Paiva Fonseca
  • Ricardo Alves Mesquita
  • Cassiano Francisco Weege Nonaka
  • Elismauro Francisco Mendonça
  • Aline Carvalho BatistaEmail author
Original Article
  • 58 Downloads

Abstract

The aim of the present study was to investigate the profile of tumor-infiltrating lymphocytes (TIL) in osteosarcomas of the jaws (OSJ). A total of 21 OSJ samples were analyzed in a retrospective and cross-sectional multicenter study. Immunohistochemistry was performed to determine the recognition of TIL such as CD4+, CD8+, granzyme B+ (GrB), programmed cell death protein+ (PD-1), and cytotoxic T lymphocyte-associated antigen 4+ (CTLA-4) in intratumoral and peripheral (stromal) regions. Positivity was determined based on the percentage and density of TIL+ per square millimeter [1 = absent (< 25 cells/mm2), 2 = low (25 to 130 cells/mm2), and 3 = high (> 130 cells/mm2)]. The association of TIL density with clinicopathologic data was determined by the Mann-Whitney test (p < 0.05). OSJ were positive for CD8+ cells in 45% (n = 9) of cases, for CD4+ cells in 30% (n = 6) of cases, and for CTLA-4+ in 4.8% (n = 1) of cases, with a score of 2 (low TIL) in all cases. All cases were negative for GrB and PD-1 (score 1). No association was observed between immune infiltrate and clinicopathologic findings. OSJ showed a microenvironment with low TIL, including failure of effectiveness of the antitumor immune response (absence of GrB+ cells), and few cells exhibited immunotherapeutic targets, such as CTLA-4 and PD-1.

Keywords

Osteosarcoma Tumor-infiltrating lymphocytes Tumor microenvironment Oral cancer Immunotherapy 

Notes

Acknowledgements

The authors thank the Coordination for the Improvement of Higher Education Personnel (CAPES), the Brazilian National Council for Scientific and Technological Development (CNPq), and the Foundation for Research Support in the State of Goiás (FAPEG). ACB, RAM, EFM, and CFWN are research fellows at CNPq. The authors thank the Coordination for the Improvement of Higher Education Personnel (CAPES). JAAA and SFSC are the recipients of fellowships. Mrs. E. Greene provided English editing of the manuscript.

Authors’ contributions

PMA, DACA, LLS, SFSC, and JAAA conducted a literature review, organized the data of the clinical cases, and conducted the immunohistochemical reactions, and morphological and immunohistochemical analysis of samples. HARP, RAM, CFWN, and EFM contributed cases from their services, and reviewed and classified, morphologically, all cases. ACB, PMA, and JAAA contributed to the design of the work. FPF was responsible for data interpretation. ACB, PMA, RAM, and HARP contributed to the conception of the work. All authors gave final approval for publication.

Compliance with ethical standards

All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Conflict of interest

All authors declare that they have no conflict of interest.

References

  1. 1.
    de Arruda JAA, Silva LVO, Kato CNAO, Schuch LF, Batista AC, Costa NL, Tarquinio SBC, Rivero ERC, Carrard VC, Martins MD, Sobral APV, Mesquita RA (2017) A multicenter study of malignant oral and maxillofacial lesions in children and adolescents. Oral Oncol 75:39–45CrossRefGoogle Scholar
  2. 2.
    Paparella ML, Olvi LG, Brandizzi D, Keszler A, Santini-Araujo E, Cabrini RL (2013) Osteosarcoma of the jaw: an analysis of a series of 74 cases. Histopathology 63(4):551–557Google Scholar
  3. 3.
    Chaudhary M, Chaudhary SD (2012) Osteosarcoma of jaws. J Oral Maxillofac Pathol 16(2):233–238CrossRefGoogle Scholar
  4. 4.
    Chen YM, Shen QC, Gokavarapu S, Ong HS, Cao W, Ji T (2016) Osteosarcoma of the mandible: a site-specific study on survival and prognostic factors. J Craniofac Surg 27(8):1929–1933CrossRefGoogle Scholar
  5. 5.
    Asioli S, Righi A, Rucci P, Tarsitano A, Marchetti C, Bacchini P, Balbi T, Bertoni F, Foschini MP (2017) p16 protein expression and correlation with clinical and pathological features in osteosarcoma of the jaws: experience of 37 cases. Head Neck 39(9):1825–1831CrossRefGoogle Scholar
  6. 6.
    Heymann MF, Lézot F, Heymann D (2017) The contribution of immune infiltrates and the local microenvironment in the pathogenesis of osteosarcoma. Cell ImmunolGoogle Scholar
  7. 7.
    Liu B, Huang Y, Sun Y, Zhang J, Yao Y, Shen Z, Xiang D, He A (2016) Prognostic value of inflammation-based scores in patients with osteosarcoma. Sci Rep 6:39862CrossRefGoogle Scholar
  8. 8.
    Liu T, Fang XC, Ding Z, Sun ZG, Sun LM, Wang YL (2015) Pre-operative lymphocyte-to-monocyte ratio as a predictor of overall survival in patients suffering from osteosarcoma. FEBS Open Bio 5:682–687CrossRefGoogle Scholar
  9. 9.
    Koirala P, Roth ME, Gill J, Piperdi S, Chinai JM, Geller DS, Hoang BH, Park A, Fremed MA, Zang X, Gorlick R (2016) Immune infiltration and PD-L1 expression in the tumor microenvironment are prognostic in osteosarcoma. Sci Rep 6:30093CrossRefGoogle Scholar
  10. 10.
    Fritzsching B, Fellenberg J, Moskovszky L, Sápi Z, Krenacs T, Machado I, Poeschl J, Lehner B, Szendrõi M, Bosch AL, Bernd L, Csóka M, Mechtersheimer G, Ewerbeck V, Kinscherf R, Kunz P (2015) CD8(+)/FOXP3(+)-ratio in osteosarcoma microenvironment separates survivors from non-survivors: a multicenter validated retrospective study. Oncoimmunology 4(3):e990800CrossRefGoogle Scholar
  11. 11.
    Pardoll DM (2012) The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12(4):252–264CrossRefGoogle Scholar
  12. 12.
    Ribas A, Wolchok JD (2018) Cancer immunotherapy using checkpoint blockade. Science 359(6382):1350–1355CrossRefGoogle Scholar
  13. 13.
    Schadendorf D, Hodi FS, Robert C, Weber JS, Margolin K, Hamid O, Patt D, Chen TT, Berman DM, Wolchok JD (2015) Pooled analysis of long-term survival data from phase II and phase III trials of ipilimumab in unresectable or metastatic melanoma. J Clin Oncol 33(17):1889–1894CrossRefGoogle Scholar
  14. 14.
    Zheng B, Ren T, Huang Y, Sun K, Wang S, Bao X, Liu K, Guo W (2018) PD-1 axis expression in musculoskeletal tumors and antitumor effect of nivolumab in osteosarcoma model of humanized mouse. J Hematol Oncol 11(1):16 Erratum in: J Hematol Oncol 2018;11(1):37CrossRefGoogle Scholar
  15. 15.
    van Erp AEM, Versleijen-Jonkers YMH, Hillebrandt-Roeffen MHS, van Houdt L, Gorris MAJ, van Dam LS, Mentzel T, Weidema ME, Savci-Heijink CD, Desar IME, Merks HHM, van Noesel MM, Shipley J, van der Graaf WTA, Flucke UE, Meyer-Wentrup FAG (2017) Expression and clinical association of programmed cell death-1, programmed death-ligand-1 and CD8(+) lymphocytes in primary sarcomas is subtype dependent. Oncotarget 8(41):71371–71384Google Scholar
  16. 16.
    Gomez-Brouchet A, Illac C, Gilhodes J, Bouvier C, Aubert S, Guinebretiere JM, Marie B, Larousserie F, Entz-Werlé N, de Pinieux G, Filleron T, Minard V, Minville V, Mascard E, Gouin F, Jimenez M, Ledeley MC, Piperno-Neumann S, Brugieres L, Rédini F (2017) CD163-positive tumor-associated macrophages and CD8-positive cytotoxic lymphocytes are powerful diagnostic markers for the therapeutic stratification of osteosarcoma patients: an immunohistochemical analysis of the biopsies fromthe French OS2006 phase 3 trial. Oncoimmunology 6(9):e1331193CrossRefGoogle Scholar
  17. 17.
    Saraf AJ, Fenger JM, Roberts RD (2018) Osteosarcoma: accelerating progress makes for a hopeful future. Front Oncol 8:4CrossRefGoogle Scholar
  18. 18.
    Costa Arantes DA, Gonçalves AS, Jham BC, Duarte ECB, de Paula ÉC, de Paula HM, Mendonça EF, Batista AC (2017) Evaluation of HLA-G, HLA-E, and PD-L1 proteins in oral osteosarcomas. Oral Surg Oral Med Oral Pathol Oral Radiol 123(6):188–196CrossRefGoogle Scholar
  19. 19.
    El-Naggar AK, Chan JKC, Grandis JR, Takata T, Slootweg PJ (2017) World Health Organization classification of head and neck tumours. IARC Press, LyonGoogle Scholar
  20. 20.
    Teng MW, Ngiow SF, Ribas A, Smyth MJ (2015) Classifying cancers based on T-cell infiltration and PD-L1. Cancer Res 75(11):2139–2145CrossRefGoogle Scholar
  21. 21.
    Webb JR, Milne K, Kroeger DR, Nelson BH (2016) PD-L1 expression is associated with tumor-infiltrating T cells and favorable prognosis in high-grade serous ovarian cancer. Gynecol Oncol 141(2):293–302CrossRefGoogle Scholar
  22. 22.
    Sundara YT, Kostine M, Cleven AH, Bovée JV, Schilham MW, Cleton-Jansen AM (2017) Increased PD-L1 and T-cell infiltration in the presence of HLA class I expression in metastatic high-grade osteosarcoma: a rationale for T-cell-based immunotherapy. Cancer Immunol Immunother 66(1):119–128CrossRefGoogle Scholar
  23. 23.
    Palmerini E, Agostinelli C, Picci P, Pileri S, Marafioti T, Lollini PL, Scotlandi K, Longhi A, Benassi MS, Ferrari S (2017) Tumoral immune-infiltrate (IF), PD-L1 expression and role of CD8/TIA-1 lymphocytes in localized osteosarcoma patients treated within protocol ISG-OS1. Oncotarget 8(67):111836–111846CrossRefGoogle Scholar
  24. 24.
    Ratti C, Botti L, Cancila V, Galvan S, Torselli I, Garofalo C, Manara MC, Bongiovanni L, Valenti CF, Burocchi A, Parenza M, Cappetti B, Sangaletti S, Tripodo C, Scotlandi K, Colombo MP, Chiodoni C (2017) Trabectedin overrides osteosarcoma differentiative block and reprograms the tumor immune environment enabling effective combination with immune checkpoint inhibitors. Clin Cancer Res 23(17):5149–5161CrossRefGoogle Scholar
  25. 25.
    Takahashi Y, Yasui T, Tamari K, Minami K, Otani K, Isohashi F, Seo Y, Kambe R, Koizumi M, Ogawa K (2017) Radiation enhanced the local and distant anti-tumor efficacy in dual immune checkpoint blockade therapy in osteosarcoma. PLoS One 12(12):e0189697CrossRefGoogle Scholar
  26. 26.
    Lussier DM, Johnson JL, Hingorani P, Blattman JN (2015) Combination immunotherapy with α-CTLA-4 and α-PD-L1 antibody blockade prevents immune escape and leads to complete control of metastatic osteosarcoma. J Immunother Cancer 3:21CrossRefGoogle Scholar
  27. 27.
    Lussier DM, O'Neill L, Nieves LM, McAfee MS, Holechek SA, Collins AW, Dickman P, Jacobsen J, Hingorani P, Blattman JN (2015) Enhanced T-cell immunity to osteosarcoma through antibody blockade of PD-1/PD-L1 interactions. J Immunother 38(3):96–106CrossRefGoogle Scholar
  28. 28.
    Burgess M, Gorantla V, Weiss K, Tawbi H (2015) Immunotherapy in sarcoma: future horizons. Curr Oncol Rep 17(11):52CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Pollianna Muniz Alves
    • 1
    • 2
  • José Alcides Almeida de Arruda
    • 3
  • Diego Antônio Costa Arantes
    • 2
  • Sara Ferreira Santos Costa
    • 3
  • Lucas Lacerda Souza
    • 4
  • Hélder Antônio Rebelo Pontes
    • 4
  • Felipe Paiva Fonseca
    • 3
  • Ricardo Alves Mesquita
    • 3
  • Cassiano Francisco Weege Nonaka
    • 1
  • Elismauro Francisco Mendonça
    • 2
  • Aline Carvalho Batista
    • 2
    • 5
    Email author
  1. 1.Department of Dentistry, School of DentistryUniversidade Estadual da ParaíbaCampina GrandeBrazil
  2. 2.Department of Stomatology (Oral Pathology), School of DentistryUniversidade Federal de GoiásGoiâniaBrazil
  3. 3.Department of Oral Surgery and Pathology, School of DentistryUniversidade Federal de Minas GeraisBelo HorizonteBrazil
  4. 4.Service of Oral Pathology, Hospital Universitário João de Barros BarretoUniversidade Federal do ParáBelémBrazil
  5. 5.Departamento de Estomatologia (Patologia Oral), Faculdade de OdontologiaUniversidade Federal de GoiásGoiâniaBrazil

Personalised recommendations