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Abstract B045: EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma

2023· article· en· W4389244706 on OpenAlexaboutno aff
Greta Jaschkowitz, Elfriede Noeßner, Angela Zaccagnino, Michael Stöckle, Kerstin Junker, Philip Zeuschner

Bibliographic record

VenueCancer Immunology Research · 2023
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicExtracellular vesicles in disease
Canadian institutionsnot available
Fundersnot available
KeywordsImmune systemCancer researchRenal cell carcinomaFlow cytometryCD8ImmunotherapyCytotoxic T cellBiologyDownregulation and upregulationCell cultureImmunologyMedicineChemistryPathologyIn vitro

Abstract

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Abstract Introduction: Treatment regimens for advanced renal cell carcinoma (RCC) involve immune checkpoint inhibition targeting the PD-(L)1 pathway. PD-L1 is often overexpressed on tumor cells in advanced disease and plays a crucial role in regulating tumor-directed immune responses. It can also be expressed on tumor-secreted extracellular vesicles (EVs), which are an essential part of intercellular communication. PD-L1 carrying EVs have been shown to have immunomodulatory effects in several tumor entities, however their role in RCC is still unknown. The aim of this study was the evaluation of putative immunomodulatory effects of RCC-derived small EVs on CD8+ T cells. Materials and Methods: EVs of 5 untreated as well as interferon γ (IFNγ)-treated RCC cell lines were isolated through serial ultracentrifugation. Quality and quantity of the EVs were verified using Western Blotting (WB), Nano Particle Tracking Analysis and Transmission Electron microscopy, their PD-L1 expression was analyzed semi quantitatively through WB. CD8+ T cells were isolated from the blood of healthy donors, activated and subsequently co-cultured with RCC-derived EVs for a duration of 3 days. Proliferation and PD-1 expression of the T cells was assessed using flow cytometry. ELISAs were performed to analyze cyto- and chemokine secretion. Results: All RCC cell lines expressed PD-L1, albeit at varying low levels. Four out of 5 treated cell lines responded to IFNγ treatment with an upregulation of PD-L1 expression on both cells and secreted EVs. Co-cultures of CD8+ T cells and RCC-derived EVs, particularly PD-L1+ EVs isolated from IFNγ-stimulated cells, lead to a reduction in T cell PD-1 expression. Additionally, PD-L1-expressing EVs of RCC cell line RCC53 had an antiproliferative effect on CD8+ T cells. EVs also increased the secretion of IL-2, IL-10, CXCL10 and CCL5 by the CD8+ T cells. Conclusions: PD-L1 is expressed at low levels in RCC cell lines. However, repetitive IFNγ stimulation can lead to upregulation of PD-L1 on both cells and EVs enabling us to use this model to further investigate EV-mediated, PD-L1 dependent effects. In co-cultures, RCC-derived EVs impede early T cell activation and proliferation patterns of CD8+ T cells. Furthermore, early results indicate a shift towards a more pro-inflammatory cyto- and chemokine production by the T cells induced by RCC-derived EVs. The results support the hypothesis of a tumor-EV-mediated immune modulation in RCC. Citation Format: Greta Jaschkowitz, Elfriede Noessner, Angela Zaccagnino, Michael Stöckle, Kerstin Junker, Philip Zeuschner. EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B045.

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How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.239
Threshold uncertainty score0.649

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0010.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.001
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.035
GPT teacher head0.336
Teacher spread0.301 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations0
Published2023
Admission routes1
Has abstractyes

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