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


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.


Osteosarcoma Tumor-infiltrating lymphocytes Tumor microenvironment Oral cancer Immunotherapy 



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.


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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

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