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An in vitro model demonstrates the potential of neoplastic human germ cells to influence the tumour microenvironment
Author(s) -
Klein B.,
Schuppe H.C.,
Bergmann M.,
Hedger M. P.,
Loveland B. E.,
Loveland K. L.
Publication year - 2017
Publication title -
andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.947
H-Index - 43
eISSN - 2047-2927
pISSN - 2047-2919
DOI - 10.1111/andr.12365
Subject(s) - peripheral blood mononuclear cell , immune system , seminoma , cytokine , biology , germ cell , immunology , t cell , cell culture , cancer research , in vitro , genetics , chemotherapy , gene
Summary Testicular germ cell tumours ( TGCT ) typically contain high numbers of infiltrating immune cells, yet the functional nature and consequences of interactions between GCNIS (germ cell neoplasia in situ) or seminoma cells and immune cells remain unknown. A co‐culture model using the seminoma‐derived TC am‐2 cell line and peripheral blood mononuclear cells ( PBMC , n = 7 healthy donors) was established to investigate how tumour and immune cells each contribute to the cytokine microenvironment associated with TGCT . Three different co‐culture approaches were employed: direct contact during culture to simulate in situ cellular interactions occurring within seminomas ( n = 9); indirect contact using well inserts to mimic GCNIS , in which a basement membrane separates the neoplastic germ cells and immune cells ( n = 3); and PBMC stimulation prior to direct contact during culture to overcome the potential lack of immune cell activation ( n = 3). Transcript levels for key cytokines in PBMC and TC am‐2 cell fractions were determined using RT ‐ qPCR . TC am‐2 cell fractions showed an immediate increase (within 24 h) in several cytokine mRNA s after direct contact with PBMC , whereas immune cell fractions did not. The high levels of interleukin‐6 ( IL 6) mRNA and protein associated with TC am‐2 cells implicate this cytokine as important to seminoma physiology. Use of PBMC s from different donors revealed a robust, repeatable pattern of changes in TC am‐2 and PBMC cytokine mRNA s, independent of potential inter‐donor variation in immune cell responsiveness. This in vitro model recapitulated previous data from clinical TGCT biopsies, revealing similar cytokine expression profiles and indicating its suitability for exploring the in vivo circumstances of TGCT . Despite the limitations of using a cell line to mimic in vivo events, these results indicate how neoplastic germ cells can directly shape the surrounding tumour microenvironment, including by influencing local immune responses. IL 6 production by seminoma cells may be a practical target for early diagnosis and/or treatment of TGCT.