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Ex vivo expansion and characterization of Tumor Infiltrating Lymphocytes from two distinct cancers – a common platform for future adoptive cell transfer therapy

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From Patients with Squamous Cell Carcinoma of the Head and Neck (SCCHN) and Malignant Melanoma (MM) it is possible to expand and preserve antitumor activity of tumor infiltrating lymphocytes (TIL) ex vivo. Clinical implementation has rendered significant success in patients with MM as published by Dudley et al. Here we show the results from the preclinical part of our project on expanding tumor active TIL from a patient with SCCHN and a patient with MM. Methods.

TIL bulk cultures are established from primary SCCHN tumors and dissiminated MM and grown in high dose IL-2. TILs are selected for high avidity and further rapidly expanded using antiCD3 and high dose IL-2.

The TIL cultures are examined for T-cell phenotype subsets, clonality and activity. Phenotyping is conducted using FACS and mAb against CD3, CD4, CD8, CD25, CD45RA, CD45RO, CCR7, CD62Ligand and CD57. Clonotypes are determined using a semiquantitative analysis of TCRBV gene segments by PCR in conjunction with T-cell receptor (TCR) clonotype mapping using denaturing gradient gel electrophoresis (DGGE). Tumor activity is tested against autologous tumor (if available), allogeneic tumor and a panel of tumor associated antigens (TAA) derived from Regulators of Apoptosis. Tumor activity is assessed with Elispot INFγ quantification. Results.

Expansion kinetics for TIL cultures showed similar results in both patients. A test rapid expansion generated up to a 3500 fold expansion of the selected TIL cultures in two to three weeks. The cultures contained mainly T effector memory cells with varying distributions of CD8+ and CD4+ subtypes among the cultures and patients. TIL from the SCCHN patient showed no response against allogeneic tumor but rather strong responses against TAAs. TIL from the MM patient showed significant allogeneic and remarkable high autologous tumor responses as well as strong TAA responses. Characterization of the clonotype composition is ongoing. Conclusions.

The procedures seems feasible for expanding tumor reactive TIL from SCCHN and MM. TIL from both patients show similar results in kinetics, phenotypes and tumor activity. Recruitment is ongoing and when further optimization of the methods has been carried out a clinical pilot study in patients with SCCHN and MM is planned.
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Document Type: Abstract

Affiliations: Center for Cancer ImmunTerapi, Hæmatologisk afd. Onkologisk afd. Herlev Hospital, Herlev Ringvej 75, 2730 Herlev

Publication date: May 1, 2008

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