The Outcome of Ex Vivo TIL Expansion Is Highly Influenced by Spatial Heterogeneity of the Tumor T-Cell Repertoire and Differences in Intrinsic In Vitro Growth Capacity between T-Cell Clones.

PURPOSE: During our efforts to develop tumor-infiltrating lymphocyte (TIL) therapy to counter the devastating recurrence rate in patients with primary resectable pancreatic ductal adenocarcinoma (PDA), we found that PDA TILs can readily be expanded in vitro and that the majority of resulting TIL cultures show reactivity against the autologous tumor. However, the fraction of tumor-reactive T cells is low. We investigated to which extent this was related to the in vitro expansion. EXPERIMENTAL DESIGN: We compared the clonal composition of TIL preparations before and after in vitro expansion using T-cell receptor (TCR) deep sequencing. Our findings for PDA were benchmarked to experiments with melanoma TILs. RESULTS: We found that the TIL TCR repertoire changes dramatically during in vitro expansion, leading to loss of tumor- dominant T-cell clones and overgrowth by newly emerging T-cell clones that are barely detectable in the tumor. These changes are primarily driven by differences in the intrinsic in vitro expansion capacity of T-cell clones. Single-cell experiments showed an association between poor proliferative capacity and expression of markers related to antigen experience and dysfunction. Furthermore, we found that spatial heterogeneity of the TIL repertoire resulted in TCR repertoires that are greatly divergent between TIL cultures derived from distant tumor samples of the same patient. CONCLUSIONS: Culture-induced changes in clonal composition are likely to affect tumor reactivity of TIL preparations. TCR deep sequencing provides important insights into the factors that govern the outcome of in vitro TIL expansion and thereby a path toward optimization of the production of TIL preparations with high therapeutic efficacy.See related commentary by Lozano-Rabella and Gros, p. 4177.

Non-contrast enhanced diagnosis of acute myocarditis based on the 17-segment heart model using 2D-feature tracking magnetic resonance imaging.

PURPOSE: The aim of this study was to investigate the diagnostic value of myocardial deformation analysis based on the 17-segment heart model using non-contrast enhanced (CE) 2D tissue feature tracking (2D-FT) technique. MATERIAL AND METHODS: Seventy patients with suspected myocarditis underwent a cardiovascular magnetic resonance (CMR) examination at 1.5 Tesla. A contrast-agent-free part of this CMR protocol was additionally performed in forty healthy volunteers (HV). Besides standard CMR data sets, 2D-FT derived segmental and global longitudinal, radial, and circumferential deformation parameters were analyzed. The 2D-FT results were compared to the combined findings from CMR imaging and endomyocardial biopsy (EMB). RESULTS: Patients were assigned to three groups depending on their ejection fraction (EF) (<40%, 40-55%, ≥55%). Compared to HV, impaired EF (<55%) was significantly correlated to reduced segmental and global strain and strain rate values. The circumferential deformation analysis was more sensitive to myocardial changes than longitudinal and radial analysis. The segmental strain/strain rate had an accuracy of 84.3%/70.0% for the diagnosis of an acute myocarditis, stated by EMB and CMR in 42 of 70 patients. In patients with preserved EF, acute myocarditis could be ruled out using only segmental strain analysis with a negative predictive value of 87.5%. CONCLUSION: In patients with suspected myocarditis, the deformation analysis based on the 17-segment heart model provides valuable information about functional myocardial inhomogeneity. This quantitative approach could be used in addition to the clinical standard CMR protocol and represents a promising tool in the framework of a prospective automatized multiparametric CMR imaging analysis.