Tumor-infiltrating lymphocytes demonstrate potent anti-tumor efficacy and synergize with PD-1 blockade in bladder cancer

Sep 22, 2026

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Bladder cancer is a prevalent urological malignancy with high recurrence and limited options for advanced disease. While immune checkpoint inhibitors provide benefit, response rates remain modest. Adoptive cell therapy using tumor-infiltrating lymphocytes (TIL) can be highly effective in other solid tumors, but its potential in bladder cancer is underexplored. Patient-derived organoids support ex vivo functional testing that preserves tumor heterogeneity.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages19

Reading Time48 min

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Abstract

Background

Methods

Results

Conclusions

Introduction

Huang et al. Journal of Translational Medicine (2026) 24:455 [https://doi.org/10.1186/s12967-026-07924-6](https://doi.org/10.1186/s12967-026-07924-6) Journal of Translational Medicine RESEARCH Open Access Tumor-infiltrating lymphocytes demonstrate potent anti-tumor efficacy and synergize with PD-1 blockade in bladder cancer Guan-Kai Huang 1,2†, Xin-Xin Zhang 1†, Lin-Yuan Huang3†, Hio-Cheng Un 1, Ren-Xuan Lin 1, Jun-Xing Chen 1*, Zong-Ren Wang 1* and Ling-Li Long4* Abstract Background Bladder cancer (BCa) is one of the most prevalent urological malignancies globally with substantial clinical challenges characterized by high recurrence rates and limited treatment options for advanced disease. Although immune checkpoint inhibitors (ICIs) provide clinical benefit, their efficacy is restricted, with objective response rates of only 20–25% . Adoptive cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) has shown remarkable success in other solid tumors but remains largely unexplored in BCa. The emergence of patient-derived organoids (PDO) offers a physiologically relevant ex vivo platform for functionally evaluating TIL by preserving tumor heterogeneity. Methods TIL were isolated from BCa patient specimens and robustly expanded ex vivo using interleukin-2 (IL-2) anda rapid expansion protocol (REP) . Expansion efficiency was quantified, and TIL phenotypes were characterized via flow cytometry. Transcriptional and clonal dynamics were profiled using single-cell RNA sequencing (scRNA-seq) coupled with T-cell receptor (TCR) sequencing. The anti-tumor efficacy of TIL, both as monotherapy and in combination with the PD-1 inhibitor Nivol umab, was assessed against BCa cell lines and autologous PDO through viability (ATP), cytokine release (IFN-γ, ELISA), and apoptosis (caspase 3/7) assays. In vivo validation was performed in a 5637 cell-linederived xenograft (CDX) mouse model. Results TIL were successfully expanded from 33 of 48 BCa samples with high yields of viable cells. Expanded TIL were predominantly CD8 +cytotoxic T lymphocytes. Integrated scRNA and TCR-seq analysis revealed that REP enriched for cytotoxic effector clones and reduced regulatory T-cell populations. TIL mediated potent tumor-killing efficacy against BCa cell lines and autologous PDO in vitro and significantly suppressed tumor growth in vivo. Critically, combining TIL †Guan-Kai Huang, Xin-Xin Zhang and Lin-Yuan Huang contributed equally to this work. *Correspondence: Jun-Xing Chen[chenjunx@mail.sysu.edu.cn](chenjunx@mail.sysu.edu.cn)[ ](chenjunx@mail.sysu.edu.cn)Zong-Ren Wang [wangzr27@mail.sysu.edu.cn](wangzr27@mail.sysu.edu.cn)[ ](wangzr27@mail.sysu.edu.cn)Ling-Li Long [longll@mail.sysu.edu.cn](longll@mail.sysu.edu.cn) Full list of author information is available at the end of the article © The Author(s) 2026. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/l](http://creativecommons.org/l)icenses/by/4.0/. Huang et al. Journal of Translational Medicine (2026) 24:455 Page 2 of 19 with Nivol umab synergistically enhanced tumor cell death in PDO and resulted in near-complete tumor suppression in the CDX model, significantly outperforming TIL monotherapy. C

FAQs

Why is bladder cancer treatment challenging despite immune checkpoint inhibitors?

Bladder cancer has substantial clinical challenges with high recurrence and limited advanced-disease options. Immune checkpoint inhibitors provide benefit but have restricted efficacy, with objective response rates around 20–25%.

How were tumor-infiltrating lymphocytes (TIL) prepared and evaluated?

TIL were isolated from bladder cancer patient specimens and expanded ex vivo using interleukin-2 with a rapid expansion protocol. Expansion efficiency and TIL phenotypes were assessed by flow cytometry, with transcriptional and clonal profiling using single-cell RNA sequencing plus T-cell receptor sequencing.

What is the effect of combining TIL with PD-1 blockade (Nivolumab)?

TIL showed potent tumor-killing activity against bladder cancer cell lines and autologous organoids in vitro and suppressed tumor growth in vivo. Combining TIL with Nivolumab synergistically enhanced tumor cell death and achieved near-complete tumor suppression in the CDX model, outperforming TIL monotherapy.

Sep 22, 2026

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