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Expansion of tumor specific Tumor-Infiltrating Lymphocytes (TIL) from sarcoma and the potential benefit of anti-CD137 stimulation: a prerequisite for Adoptive Cell Transfer (ACT) immunotherapy (14PD)

Date

09 Dec 2017

Session

Poster Discussion session

Presenters

Morten Nielsen

Citation

Annals of Oncology (2017) 28 (suppl_11): xi3-xi5. 10.1093/annonc/mdx710

Authors

M. Nielsen1, A. Krarup-Hansen2, D. Hovgaard3, M.M. Petersen3, A.C. Loya4, M.C.W. Westergaard1, I.M. Svane1, N. Junker2

Author affiliations

  • 1 Center For Cancer Immune Therapy, Department Of Haematology And Department Of Oncology, Herlev and Gentofte Hospital, 2730 - Herlev/DK
  • 2 Department Of Oncology, Herlev and Gentofte Hospital, 2730 - Herlev/DK
  • 3 Department Of Orthopedic Surgery, Rigshospitalet, University of Copenhagen, 2100 - Copenhagen/DK
  • 4 Department Of Pathology, Rigshospitalet, University of Copenhagen, 2100 - Copenhagen/DK
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Background

Tumor specific TIL can be in vitro expanded and have the ability to induce complete and durable tumor regression in some patients with melanoma following ACT. In this preclinical study, we investigated the feasibility of expanding TIL from sarcomas, as well as performing functional in vitro analyses on these.

Methods

Fresh tumor samples were obtained, and TIL were isolated and expanded in growth medium containing IL-2. In a sub study, we investigated the effect of adding an agonistic CD137 antibody (Urelumab, BMS) and/or an anti-CD3 antibody (OKT3) to the medium. Phenotype and functional analyses was performed using flow cytometry and IFNγ-Elispot. Cytotoxicity analyses were performed using Xcelligence.

Results

Tumor samples from 28 patients with 8 different sarcoma subtypes were obtained, and we were able to expand a minimum of 40 million TILs from 25 of these. Mean expansion times were 32 days (14 - 61). 87,7% (36,4 – 99,1) of these cells were CD3+, and of these, 66,7% (16,3 – 99,1) were CD4+, and 21,8% (0,1 – 50,6) were CD8+. Adding anti-CD137 and/or OKT3 increased total yield of TILs; anti-CD137 skewed the phenotype significantly towards more CD8+ TILs and in some cases NK cells and γδ cells. TILs from 11 of 22 tested tumor samples from 7 of 8 different sarcoma subtypes demonstrated reactivity against autologous tumor cells using IFNγ-Elispot. These results were verified in an intracellular cytokine release assay using flow cytometry, and in cytotoxicity assays. In some cases the fraction of reactive cells was more than 20%. In TILs stimulated with anti-CD137 the reactivity increased in four of four tested samples.

Conclusions

We were able to expand TIL from 90% of tested tumor samples. Expanded TIL were a mix of CD4+ and CD8+ with CD4+ being predominant. Half of the TIL cultures showed in vitro tumor reactivity, which in some cases was as high as previously only seen in melanoma samples. Early analyses suggest that addition of anti-CD137 could influence expansion time, phenotype, and functional capacity of the expanded TIL. Based on these results, we plan to initiate clinical testing of TIL based ACT in sarcoma patients.

Clinical trial identification

Legal entity responsible for the study

Center for Cancer Immune Therapy, Herlev Hospital

Funding

Center for Cancer Immune Therapy, Herlev Hospital

Disclosure

All authors have declared no conflicts of interest.

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