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Cytokine screening identifies TNF to potentially enhance immunogenicity of pediatric sarcomas
Hendrik Gassmann1,2, Melanie Thiede1, Jennifer Weiß1
1Department of Pediatrics, Children's Cancer Research Center, Kinderklinik München Schwabing, TUM School of Medicine, Technical University of Munich, Munich, Germany.
Introduction:
Pediatric sarcomas, including osteosarcoma (OS), Ewing sarcoma (EwS) and rhabdomyosarcoma (RMS) carry low somatic mutational burden and low MHC-I expression, posing a challenge for T cell therapies. Our previous study showed that mediators of monocyte maturation sensitized the EwS cell line A673 to lysis by HLA-A*02:01/CHM1319-specific allorestricted T cell receptor (TCR) transgenic CD8+ T cells (CHM1319 CD8+ T cells).
Methods:
In this study, we tested a panel of monocyte maturation cytokines for their ability to upregulate immunogenic cell surface markers on OS, EwS and RMS cell lines, using flow cytometry. xCELLigence, SRB and ELISpot assays were used to assess whether TNF pretreatment increases CD8+ T cell cytotoxicity.
Results:
We observed that TNF and IL-1β upregulated MHC class I, ICAM-1 as well as CD83 and PD-L1 on the surface of pediatric sarcoma cell lines, while IL-4, GM-CSF, IL-6 and PGE2 failed to induce respective effects. Although pretreatment of pediatric sarcoma cell lines with TNF did not improve unspecific peripheral blood mononuclear cells (PBMCs) cytotoxicity, TNF enhanced specific lysis of 1/3 HLA-A2+ EwS cell lines by CHM1319 CD8+ T cells depending on MHC-I expression and ICAM-1 upregulation.
Discussion:
Our study supports utilization of TNF or TNF-inducing regimens for upregulation of MHC-I and costimulatory surface molecules on pediatric sarcoma cells and for enhancing recognition of responsive HLA-A2+ EwS tumor cells by antigen-specific CD8+ T cells.
Insights
Tumor necrosis factor (TNF) and IL-1β increase immune markers on pediatric sarcoma cells. This enhances T cell therapy effectiveness against specific Ewing sarcoma tumors expressing HLA-A2.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Pediatric sarcomas (osteosarcoma, Ewing sarcoma, rhabdomyosarcoma) have low mutation burden and MHC-I expression, hindering T cell therapies.
- Previous research indicated monocyte maturation mediators sensitize Ewing sarcoma to T cell-mediated lysis.
Purpose of the Study:
- To evaluate cytokines' ability to upregulate immunogenic markers on pediatric sarcoma cell lines.
- To determine if TNF pretreatment enhances CD8+ T cell cytotoxicity against these cells.
Main Methods:
- Flow cytometry was used to assess cell surface marker expression.
- xCELLigence, SRB, and ELISpot assays evaluated T cell cytotoxicity after TNF pretreatment.
- Tested cytokines included TNF, IL-1β, IL-4, GM-CSF, IL-6, and PGE2.
Main Results:
- TNF and IL-1β upregulated MHC class I, ICAM-1, CD83, and PD-L1 on pediatric sarcoma cell lines.
- Other tested cytokines did not induce these effects.
- TNF enhanced specific lysis of HLA-A2+ Ewing sarcoma cells by antigen-specific T cells, dependent on MHC-I and ICAM-1.
Conclusions:
- TNF or TNF-inducing regimens can upregulate key surface molecules on pediatric sarcoma cells.
- This approach enhances recognition of HLA-A2+ Ewing sarcoma cells by antigen-specific CD8+ T cells, improving T cell therapy potential.
