Isolation of tumor-resident CD8+ T cells from human lung tumors

Corgnac, Stéphanie; Mami‐Chouaib, Fathia

doi:10.1016/j.xpro.2020.100267

Cited by 6 publications

(6 citation statements)

References 5 publications

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“…To exclude an artifact due to allogenic reaction, we aimed to confirm this finding in fully autologous ToC co-cultures. We developed a strategy to freshly isolate the three cell types (cancer cells, CD8 + CTLs, and CAFs) from surgical NSCLC tumor samples by sequential magnetic cell sorting (MACS) 33 and to shortly amplify them ex vivo before ToC generation ( Figure 5 A). Each cell type was expanded for 3 weeks in its appropriate medium and growth support: CD8 + cells in suspension in V-bottom multiwell plates, cancer cells as spheroids in ultra-low attachment flasks, and adherent CAFs in Petri dishes.…”

Section: Resultsmentioning

confidence: 99%

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Veith,

Nurmik,

Mencattini

et al. 2024

Cell Reports Medicine

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Section: Resultsmentioning

confidence: 99%

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Veith,

Nurmik,

Mencattini

et al. 2024

Cell Reports Medicine

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“…Stained cells were analyzed by flow cytometry using a BD FACS Fortessa flow cytometer (BD Biosciences). Data were processed using FlowJo software (Tree Star Inc..) (Corgnac, Lecluse et al, 2021). This study was approved by the Institutional Review Board of Gustave Roussy (Commission scientifique des Essais thérapeutiques [CSET]) and informed consent was obtained.…”

Section: Methodsmentioning

confidence: 99%

Differential predictive value of resident memory CD8⁺T cell subpopulations in non-small-cell lung cancer patients treated by immunotherapy

Paolini,

Tran,

Corgnac

et al. 2024

Preprint

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A high density of resident memory T cells (TRM) in tumors correlates with improved clinical outcomes in immunotherapy-treated patients. However, in preclinical models, only some subpopulations of TRM are associated with cancer vaccine efficacy. We identified two main TRM subpopulations in tumor-infiltrating lymphocytes derived from non-small cell lung cancer (NSCLC) patients: one co-expressing CD103 and CD49a (DP), and the other expressing only CD49a (MP); both exhibiting additional TRM surface markers like CD69. DP TRM exhibited greater functionality compared to MP TRM. Analysis of T-cell receptor (TCR) repertoire and of the stemness marker TCF-1 revealed shared TCRs between populations, with the MP subset appearing more progenitor-like phenotype. In two NSCLC patient cohorts, only DP TRM predicted PD-1 blockade response. Multivariate analysis, including various biomarkers (CD8, TCF1+CD8+T cells, and PD-L1) associated with responses to anti-PD(L)1, showed that only intra-tumoral infiltration by DP TRM remained significant. This study highlights the non-equivalence of TRM populations and emphasizes the importance of distinguishing between them to better define their role in antitumor immunity and as a biomarker of response to immunotherapy.

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“…The tumor was collected and processed in our laboratory. Tumor dissociation was adapted from Corgnac et al [ 9 ]. Upon arrival, the tumor tissues were weighed and washed with phosphate-buffered saline.…”

Section: Methodsmentioning

confidence: 99%

“…We sorted with magnetic beads tumor cells (Miltenyi, 130-108-339) and CD8+ lymphocytes (Miltenyi, 130-121-560) from the primary tumor. Then we loaded tumor cells at 2 × 10 6 cells/mL in the chip, left them self-organize for 48 h, and then added a second droplet including the CD8+ population according to the protocol [ 9 ]. We cultured the CD8+ cells during that 48 h with the RPMI 1640 (Gibco, 61870-010) with 10% serum human group AB (Biotechnology Institute Jacques Boy, France, 201021334), 1% sodium pyruvate (Gibco, 11360-039), 0.2% penicilline/streptavidine filtered with Millipore 0.22 μm size (Sigma, Saint-Quentin-Fallavier France, S2GPT01RE) supplemented with 50 units of interleukin-2 (IL-2, Miltenyi Biotec, 130-097-744).…”

Section: Methodsmentioning

confidence: 99%

Evaluating immune response in vitro in a relevant microenvironment: a high-throughput microfluidic model for clinical screening

Doffe

Fuoco

Michels

et al. 2022

Exploration of Targeted Anti-Tumor Therapy

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Aim: Functional screening of new pharmaceutical compounds requires clinically relevant models to monitor essential cellular and immune responses during cancer progression, with or without treatment. Beyond survival, the emergence of resistant tumor cell clones should also be considered, including specific properties related to plasticity, such as invasiveness, stemness, escape from programmed cell death, and immune response. Numerous pathways are involved in these processes. Defining the relevant ones in the context of a specific tumor type will be key to designing an appropriate combination of inhibitors. However, the diversity and potential redundancy of these pathways remain a challenge for therapy. Methods: A new microfluidic device developed by Okomera was dedicated to the screening of drug treatment for breast cancer. This microchip includes 150 droplet-trapping microwells, offering multi-chip settings and multiple treatment choices. Results: After validating the system with established cell lines and a panel of drugs used clinically at Gustave Roussy, preclinical experiments were initiated including patient-derived xenograft (PDX) and primary tumor cells-derived tumoroids with the collaboration of Gustave Roussy clinicians. Tumor-isolated lymphocytes were also added to the tumoroids, using secondary droplets in proof-of-concept experiments. Conclusions: These results show the relevance of the methodology for screening large numbers of drugs, a wide range of doses, and multiple drug combinations. This methodology will be used for two purposes: 1) new drug screening from the compound library, using the high throughput potential of the chip; and 2) pre-clinical assay for a two-weeks response for personalized medicine, allowing evaluation of drug combinations to flag an optimized treatment with potential clinical application.

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Isolation of tumor-resident CD8+ T cells from human lung tumors

Cited by 6 publications

References 5 publications

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Differential predictive value of resident memory CD8⁺T cell subpopulations in non-small-cell lung cancer patients treated by immunotherapy

Evaluating immune response in vitro in a relevant microenvironment: a high-throughput microfluidic model for clinical screening

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Isolation of tumor-resident CD8+ T cells from human lung tumors

Cited by 6 publications

References 5 publications

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Assessing personalized responses to anti-PD-1 treatment using patient-derived lung tumor-on-chip

Differential predictive value of resident memory CD8+T cell subpopulations in non-small-cell lung cancer patients treated by immunotherapy

Evaluating immune response in vitro in a relevant microenvironment: a high-throughput microfluidic model for clinical screening

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Differential predictive value of resident memory CD8⁺T cell subpopulations in non-small-cell lung cancer patients treated by immunotherapy