bims-tuinly 2026-11-01 papers

bims-tuinly

Biomed News

on Tumor-infiltrating lymphocytes therapy

Issue of 2026–11–01
24 papers selected by
Pierpaolo Ginefra, Ludwig Institute for Cancer Research

LM103, an autologous tumor-infiltrating lymphocyte monotherapy, in patients with advanced melanoma resistant to standard therapy.
Human platelet lysate enables robust expansion of tumor infiltrating lymphocytes from endometrial cancer.
Tumour-infiltrating lymphocytes (TILs) in oral cavity squamous cell carcinoma: Histopathological analysis and its correlation with clinicopathological parameters.
Treatment-specific Association of H&E-based Stromal Tumor-infiltrating Lymphocytes With Survival in Pleural Mesothelioma Treated With Ipilimumab/Nivolumab or Chemotherapy.
Clinical-scale manufacturing of CD137+ enriched tumor-reactive T cells for glioblastoma in the CliniMACS Prodigy.
Histopathological and Clinical Perspectives on Prognostic Markers in Endometrial Cancer.
Cancer-associated fibroblasts shape features of lymphocyte aggregation and collagen deposition in hepatocellular carcinoma.
Integration of TRM-Cell Infiltration and PD-L1 Expression Improves Prognostic Stratification After Resection of NSCLC.
Predicting recurrence risk after neoadjuvant therapy in breast cancer: a single-center retrospective cohort study integrating residual cancer burden and immune microenvironment features.
hnRNPA2B1 sustains antitumor T cell immunity and enhances cancer immunotherapy through IDR-dependent stabilization of chromatin-regulatory transcripts.
Methods for Analyzing Immune Infiltrates and Spatial Organization Within the Melanoma Microenvironment Using Multiplex Immunofluorescence.
Reprogramming the Melanoma Tumor Microenvironment: From Cellular Crosstalk to Cellular Immunotherapy.
The Y chromosome gene KDM5D restrains CD8+ T cell antitumor immunity through TCR and cholesterol-exhaustion programs.
Tumor Immune Microenvironment and VAF-inferred BRCA1/2 Loss of Heterozygosity in Japanese Patients With Germline BRCA1/2-mutated Breast Cancer.
Programmable antigen-presenting scaffolds expand rare tumor-antigen-specific T cells from peripheral blood.
Generation of Murine Chimeric Antigen Receptor T Cells for Adoptive T Cell Therapy for Melanoma.
Time-Encoded Ligand Presentation via a Magnetically Actuated Hydrogel Mitigates T Cell Exhaustion.
Antibody Targeting Soluble NKG2D Ligand MIC Enhances Adoptive T-Cell Therapy in an NK Cell-Dependent Mechanism.
Immune- and Stromal-Associated Tissue-Context Profiles in Pretreatment Triple-Negative Breast Cancer: An Exploratory Institutional Cohort Study.
Clinicopathological, Molecular, and Immune Profiling in Colorectal Cancer: Associations Between KRAS/BRAF Alterations, dMMR/MSI-H Status, and Tumor-Infiltrating Lymphocytes: A Retrospective Cross-Sectional Study.
Experimental high blood glucose metabolically rejuvenates CD8+ T cells for improved control of tumor growth through an IGF1-IGF1R axis.
Overcoming the efficacy-safety dilemma of cytokine therapy for tissue-resident memory T cells in cancer immunotherapy: Intrinsic sensitization and extrinsic optimization.
A Method for Quantifying Immune Infiltrates and Tertiary Lymphoid Structures in Murine Melanoma Tumors by Multiplex Immunofluorescence.
Association of pretreatment immune microenvironment profile with boron neutron capture therapy efficacy in cutaneous tumors: results from an ancillary multiplex immunofluorescence study.

Cancer Immunol Res. 2026 Oct 23.

LM103, an autologous tumor-infiltrating lymphocyte monotherapy, in patients with advanced melanoma resistant to standard therapy.

Li Zhou, Xiaojun Cai, Haizhen Du, Xuan Wang, Yue Yang, Zhichao Tan, Jiayong Liu, Zhihong Chi, Lu Si, Lili Mao, Bin Lian, Hui Tian, Qian Guo, Hua Wang, Ming Ji, Lulu Wang, Yuan Yuan, Yongming Xue, Jun Guo, Yan Kong, Chuanliang Cui.

We conducted a phase I study to evaluate the safety, clinical activity, and immune correlates of autologous tumor-infiltrating lymphocytes (TIL) therapy LM103 in Chinese patients with refractory metastatic melanoma. Fourteen patients with refractory metastatic melanoma received lymphodepleting chemotherapy followed by LM103 infusion (3.58-19.47×10¹⁰ cells) and high-dose interleukin-2. Characteristics of baseline tumor, TIL products and pre-/post-infusion peripheral blood were analyzed. LM103 therapy demonstrated manageable safety and promising efficacy. TEAEs included myelosuppression (100%), capillary leak syndrome (100%), fever (85.7%), and rash (71.4%). Grade 3-4 TEAEs included leukopenia (85.7%), neutropenia (78.6%), lymphopenia (78.6%), fever (71.4%), and thrombocytopenia (57.1%). The objective response rate was 35.7% (95% confidence interval [CI], 12.76 to 64.86), and the disease control rate was 85.7% (95% CI, 57.19 to 98.22). Median progression-free survival was 4.52 months (95% CI, 2.07 to 14.31), and median overall survival was 16.58 months (95% CI, 5.16 to not estimable). Baseline tumor immune infiltration and mutational burden were not associated with clinical outcome. LM103 infusion induced marked peripheral T cell receptor repertoire remodeling, with more than 60% of infused TIL clones detectable in peripheral blood at 6 months. Patients with superior clinical outcomes demonstrated greater expansion and persistence of dominant TIL clones. In addition, genes favoring T cells tumor infiltration were upregulated in TILs of patients with longer PFS. LM103 showed manageable safety and encouraging antitumor activity. Clinical benefit was associated with sustained in vivo clonal persistence and upregulated genes favoring tumor re-infiltration of infused TILs. Trial registration number CTR20233999.

DOI: https://doi.org/10.1158/2326-6066.CIR-26-0591
Front Immunol. 2026 ;17 1873651

Human platelet lysate enables robust expansion of tumor infiltrating lymphocytes from endometrial cancer.

Willemiek Iwema, Nienke van Rooij, Lena Rockstein, Annechien Plat, Refika Yigit, Joost Bart, Bahez Gareb, Hans W Nijman, Edwin Bremer, Marco de Bruyn.

   Background/objectives: Tumor infiltrating lymphocytes (TIL) therapy has yielded promising results in melanoma patients, with an approved product available. TIL therapy is of similar interest to a number of additional malignancies, including endometrial cancer (EC). For EC specifically, the tumor is well infiltrated with TILs and has accessible tumors as starting material for TIL isolation/expansion. However, the TIL manufacturing process currently relies on human serum (HS), which is difficult to source and has considerable batch-to-batch variation. Here, we explored the feasibility of using human platelet lysate (HPL) as a good manufacturing practice (GMP)-compatible alternative.
Methods: EC material was obtained following surgery and used to expand TILs in pre-rapid expansion protocols (pre-REP) and REP phases, comparing HPL- and HS-containing media. Tumor tissue from 10 EC patients, representing both high and low tumor mutational burden histotypes was used. Spatial TIL infiltration was assessed using immunohistochemistry. After expansion, TIL phenotype was determined using flow cytometry for CD3, CD56, CD8, CD39, and CD103. TILs from the REP were further characterized with spectral flow cytometry using markers for differentiation, activation, co-stimulatory receptors, and immune checkpoints.
Results: TILs expanded comparably using both HPL- and HS-media during pre-REP and REP in terms of cell yields. All tumors showed an excluded spatial immune phenotype. Immunophenotypes were consistent across media conditions. Spectral cytometry revealed patient-specific clustering unaffected by supplement type.
Conclusions: TILs can be successfully expanded from EC using a fully GMP-compatible HPL protocol. Our findings may serve as a foundation for further improvements of existing GMP-compliant TIL manufacturing protocols.

Keywords:  GMP manufacture; endometrial cancer; human platelet lysate; human serum; tumor infiltrating lymphocytes

DOI:  https://doi.org/10.3389/fimmu.2026.1873651
Indian J Pathol Microbiol. 2026 Jul 01. 69(3): 455-459

Tumour-infiltrating lymphocytes (TILs) in oral cavity squamous cell carcinoma: Histopathological analysis and its correlation with clinicopathological parameters.

Divya Sharma, Nishta Gupta, Anand Kumar Verma.

   BACKGROUND: Tumor-infiltrating lymphocytes (TILs) reflect the immune system's response to cancer and their significance in Oral cavity Squamous Cell Carcinoma (OCSCC) is being increasingly recognized.
AIM: This study is aimed to evaluate the histopathological presence and distribution of TILs in OCSCC and correlate them with clinicopathological parameters such as tumor grade, stage, perineural invasion (PNI) and lymphovascular invasion (LVI).
MATERIAL AND METHODS: A cross-sectional observational study was conducted on 41 patients with OCSCC using tissue samples stained with hematoxylin and eosin. TILs were quantified in stromal and intratumoral regions and statistically correlated with clinical parameters using SPSS version 26.
RESULTS: High stromal TILs were observed in 90.2% of patients, while only 17% showed high intratumoral TILs. A significant inverse association was observed between TIL scores and tumor stage. Also statistical significance was noted between PNI and low stromal TILs.
CONCLUSION: Morphological assessment of TILs using the H and E-stained sections can aid in immune-based classification and prognostication of OCSCC. TIL scoring can be incorporated into clinicopathological prognostic models and can be further explored for targeted immune therapies in OCSCC.

Keywords:  Oral cavity squamous cell carcinoma; tumor microenvironment; tumor-infiltrating lymphocytes (stromal and intra tumoral)

DOI:  https://doi.org/10.4103/ijpm.ijpm_782_25
Anticancer Res. 2026 Nov;46(11): 6193-6202

Treatment-specific Association of H&E-based Stromal Tumor-infiltrating Lymphocytes With Survival in Pleural Mesothelioma Treated With Ipilimumab/Nivolumab or Chemotherapy.

Takashi Yamasaki, Koji Yamanegi, Hiroaki Ota, Daichi Fujimoto, Mai Nakanishi, Soichiro Funaki, Takashi Kijima, Chisato Ohe.

   BACKGROUND/AIM: Pleural mesothelioma lacks robust biomarkers to guide personalized treatment selection. Although stromal tumor-infiltrating lymphocytes (sTILs) on hematoxylin and eosin (H&E)-stained sections represent a cost-effective biomarker in various cancers, their clinical significance in pleural mesothelioma remains unclear. This study evaluated the prognostic impact of sTILs in pleural mesothelioma patients treated with platinum-pemetrexed chemotherapy or dual immune checkpoint inhibition (ICI) using ipilimumab plus nivolumab (Ipi/Nivo).
PATIENTS AND METHODS: We retrospectively analyzed 179 patients with unresectable pleural mesothelioma who received either platinum-pemetrexed chemotherapy (n=127) or Ipi/Nivo (n=52). sTILs were assessed according to the International TILs Working Group guidelines and categorized as high (>10%) or low (≤10%). Overall survival (OS) and associations between sTIL status and clinicopathological characteristics were evaluated.
RESULTS: High and low sTIL status was observed in 82 (45.8%) and 97 (54.2%) patients, respectively. Within each treatment cohort, baseline clinicopathological characteristics were generally well-balanced according to sTIL status. In the entire cohort, sTIL status was not significantly associated with OS (p=0.559). Subgroup analyses according to treatment modality showed no significant association between sTIL status and OS in the platinum-pemetrexed cohort (p=0.664). In contrast, in the Ipi/Nivo cohort, OS was significantly longer in the low sTIL group than in the high sTIL group (p=0.027).
CONCLUSION: sTILs may have treatment-specific prognostic significance in pleural mesothelioma. Notably, low sTIL status was associated with longer survival among patients treated with dual ICI but not chemotherapy. Standardized H&E-based sTIL assessment may provide a simple and accessible approach for refining treatment selection in pleural mesothelioma.

Keywords:  Pleural mesothelioma; chemotherapy; immunotherapy; prognostic prediction; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.21873/anticanres.18449
Front Immunol. 2026 ;17 1866645

Clinical-scale manufacturing of CD137+ enriched tumor-reactive T cells for glioblastoma in the CliniMACS Prodigy.

Arina Riabinska, Elvira Criado-Moronati, Emre Balta, Denis Frison, Cristina Berganza-Irusqueta, Tümay Telatar, Donata Maciaczyk, Johanna Engeln, Laura Fischer, Marieke Domdey, Imran Siddiqui, Velita Miranda, Teresa Schmidt, Sied Kebir, Yahya Ahmadipour, Martin Glas, Ulrich Herrlinger, Matthias Schneider, Anna-Laura Potthoff, Marcus Czabanka, Thomas Broggini, Sofia Stenqvist, Lisa M Nilsson, Jonas A Nilsson, Martina Ott, Andrzej Dzionek, Ian R Hardy.

   Background: Most tumor-infiltrating lymphocyte (TIL) studies and therapies have focused on tumors with high T-cell infiltration and mutational burden. Glioblastoma (GBM) represents a challenging indication for TIL therapy because of its limited T-cell infiltration and low mutational burden. We investigated whether targeted enrichment of tumor-reactive T cells using CD137-based selection could enable robust, clinical-scale production of TIL products from GBM.
Methods: Ultrasonic aspirates (UAs), routinely collected during GBM resections, were evaluated as a source of TILs for clinical manufacturing. TILs were generated from GBM UAs under GMP-like conditions at clinical scale using the closed, automated TRT Process on the CliniMACS Prodigy. Matched TRT products that were enriched for CD137+ cells, depleted of CD137+ cells, or left unselected were generated in parallel at full manufacturing scale. Products were characterized by cellular and transcriptomic analyses and evaluated in functional assays for cytotoxicity against autologous and non-autologous patient-derived GBM cell lines.
Results: Cellular and transcriptomic analyses revealed that UA-derived CD137+ (4-1BB/TNFRSF9) T cells were clonally expanded, exhibited activated and memory phenotypes, and displayed transcriptional profiles associated with tumor reactivity. The closed manufacturing process preserved phenotypic features previously associated with clinical benefit in other TIL indications. Functional assays demonstrated potent, tumor-reactive cytotoxicity of CD137-enriched TRT products against autologous GBM cell lines compared with TILs generated in parallel without selection.
Discussion/Conclusions: These findings establish the feasibility of a clinical-scale TIL manufacturing approach using the CliniMACS Prodigy that enables production of clinically relevant quantities of TILs from GBM while enriching for cells with tumor-reactive characteristics. This approach may broaden the application of TIL therapy to immunologically cold tumors with limited T-cell infiltration or low mutational burden that have historically been challenging to manufacture using conventional TIL production methods.

Keywords:  CD137+ T cells; CliniMACS Prodigy®; T cell therapy; glioblastoma; tumor infiltrating lymphocyte (TIL)

DOI:  https://doi.org/10.3389/fimmu.2026.1866645
Int J Gynecol Pathol. 2026 Oct 28.

Histopathological and Clinical Perspectives on Prognostic Markers in Endometrial Cancer.

Figen Aslan, Gözde Kurtuluş, Demet Açıkalın, Hülya Kaçar, Duygu Kavak, Nadi Keskin.

  The microcystic, elongated, and fragmented (MELF) invasion pattern, tumor budding (TB), and tumor-infiltrating lymphocytes (TILs) may have prognostic value in endometrial carcinoma; however, their relationships are not fully defined. This retrospective study included 216 endometrioid endometrial carcinomas. MELF pattern, TB (BD1-3), and TIL levels were evaluated on hematoxylin-eosin sections. Clinicopathologic variables and immunohistochemical markers (HER2, p53, mismatch repair proteins) were analyzed statistically, including survival analysis.MELF pattern of invasion (present in 32.4%) was significantly associated with larger tumor size, low-to-intermediate grade, deep myometrial invasion, lymphovascular invasion (LVI), and nodal metastasis; on multivariate analysis, it remained independently associated with LVI (odds ratio=38.15) and nodal metastasis (odds ratio=12.82). TB was significantly associated with high-grade, deep invasion, cervical involvement, advanced FIGO stage, and mortality after Bonferroni correction; associations with tumor size, LVI, and nodal metastasis were significant only at the conventional threshold and considered trends. TB did not retain independent prognostic significance in the Cox regression model. TIL showed no significant association with any parameter, survival, or the other 2 patterns. MELF pattern of invasion is a clinically important marker independently associated with LVI and nodal metastasis. Notably, MELF was more common in low-to-intermediate grade tumors, suggesting that it may identify aggressive invasive behavior in tumors otherwise considered lower risk by grade alone. TB reflects aggressive tumor behavior without independent survival significance. TIL had limited prognostic value by routine assessment. Combined evaluation of MELF and TB may improve risk stratification in endometrioid endometrial carcinoma, particularly where molecular profiling is unavailable.

Keywords:  Endometrioid endometrial carcinoma; Lymphovascular invasion; MELF invasion pattern; Survival; Tumor budding; Tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.1097/PGP.0000000000001224
J Transl Med. 2026 Jul 30. pii: 1353. [Epub ahead of print]24(1):

Cancer-associated fibroblasts shape features of lymphocyte aggregation and collagen deposition in hepatocellular carcinoma.

Yuan'e Lian, Zehong Zhang, Dongsheng Xia, Zhongchang Weng, Jiaying Ye, Siqin Zhang, Xingxing Huang, Lianhuang Li, Jie Lin, Songqiang Zhou, Shi Chen, Yannan Bai.

BACKGROUND: The progression of hepatocellular carcinoma (HCC) is significantly influenced by the tumor microenvironment, in which cancer-associated fibroblasts (CAFs), collagen fibers (CFs), and tumor-infiltrating lymphocytes (TILs) interact to regulate tumor growth and immune infiltration. Understanding the spatial and functional coordination among these components is essential for prognostic assessment.
METHODS: This retrospective study included 335 patients from two tertiary hospitals. Early-stage HCC tissue samples were analyzed using artificial intelligence-based algorithms to quantify TIL fraction and aggregation, while collagen architecture was characterized using multiphoton microscopy (MPM). Moreover, single-cell RNA sequencing was used for functional profiling of CAFs, and multiplex immunofluorescence was applied for spatial validation. The relationship between TILs and collagen features was associated with clinical outcomes. Prognostic models were developed using Cox regression and validated internally and externally.
RESULTS: MPM demonstrated strong co-localization of CFs and TILs, with lymphocytes spatially aligned following collagen orientation within the tumor core. CAF subtypes exhibited heterogeneous expression of extracellular matrix (ECM)-related and lymphocyte-modulatory genes, with ECM-mediated signaling dominating CAF-T/B cell communication. TIL aggregation and CF orientation independently predicted disease-free and overall survival. The multifactorial Fiber-TIL-Clinical model (FTC model) achieved superior predictive performance (area under the curve = 0.902 for early recurrence) by integrating these features with clinical variables.
CONCLUSIONS: The FTC model integrates CF features and TIL characteristics, demonstrating strong performance in predicting HCC recurrence.

DOI: https://doi.org/10.1186/s12967-026-08686-x
Anticancer Res. 2026 Nov;46(11): 6307-6318

Integration of TRM-Cell Infiltration and PD-L1 Expression Improves Prognostic Stratification After Resection of NSCLC.

Takuya Tanimura, Masanori Shiohara, Ayaho Ueno, Kazuya Kishimoto, Hirotaka Kinoshita, Satoshi Suzuki, Hiroaki Nagamine, Kantaro Hara, Hidetoshi Inoue, Shinjiro Mizuguchi, Hiroyasu Kaneda, Masakazu Yashiro, Kenichi Kohashi, Junichi Soh.

   BACKGROUND/AIM: Tumor-infiltrating lymphocytes, including tissue-resident memory T (TRM) cells, are associated with prognosis in patients after resection of non-small-cell lung cancer (NSCLC). However, their clinical significance in the context of tumor immune suppression, such as through expression of programmed cell death ligand-1 (PD-L1), remains unclear. We hypothesized that integrating TRM-cell infiltration with PD-L1 expression would improve prognostic stratification.
PATIENTS AND METHODS: We retrospectively analyzed 316 patients with NSCLC who underwent curative resection between 2017 and 2020 without perioperative immunotherapy. TRM cells were quantified by CD103 immunohistochemistry using tissue microarrays and digitally analyzed with QuPath. PD-L1 expression was assessed using the 22C3 assay. Recurrence was evaluated using competing-risks analysis. Recurrence-free survival (RFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox proportional hazards models.
RESULTS: The median TRM density was 54.8 cells/mm2 and was used to define TRM-high and TRM-low groups. TRM-high tumors were associated with non-adenocarcinoma histology and PD-L1 expression ≥1%. Patients with TRM-high tumors showed a significantly lower cumulative incidence of recurrence (p=0.039), whereas PD-L1 expression ≥1% was associated with worse RFS and OS. When combined, TRM-high/PD-L1 expression <1% demonstrated lowest cumulative incidence of recurrence (p=0.017) and significantly improved RFS and OS (p=0.0035 and p=0.0031, respectively). In multivariable analysis, TRM-high/PD-L1 expression <1% was independently associated with lower recurrence (hazard ratio=0.36) and improved RFS (hazard ratio=0.47).
CONCLUSION: The combined assessment of TRM-cell infiltration and PD-L1 expression provides robust prognostic stratification in resected NSCLC and may serve as a useful biomarker for predicting postoperative outcomes.

Keywords:  CD103; NSCLC; PD-L1; TRM cells; Tissue-resident memory T-cells; non-small-cell lung carcinoma; programmed cell death ligand-1; tumor immune microenvironment

DOI:  https://doi.org/10.21873/anticanres.18459
Front Oncol. 2026 ;16 1925326

Predicting recurrence risk after neoadjuvant therapy in breast cancer: a single-center retrospective cohort study integrating residual cancer burden and immune microenvironment features.

Zhihua Zhao, Rui Deng, Liangliang Bai, Xiaogang Duan, Jingyuan Yi.

   Background: Residual cancer burden (RCB) after neoadjuvant therapy (NAT) provides prognostic information, but its combination with immune microenvironment features for recurrence prediction remains underexplored.
Methods: This single-center retrospective cohort study included 420 consecutive patients with breast cancer who underwent neoadjuvant therapy followed by definitive surgery between January 2015 and June 2024. Residual cancer burden (RCB) and post-neoadjuvant immune features, including tumor-infiltrating lymphocytes (TILs), CD8, FOXP3, and the proportion of CD163-positive among CD68-positive macrophages, were assessed. Patients were stratified by calendar era and randomly partitioned within strata into a training cohort (n=294) and an internal hold-out validation cohort (n=126). A LASSO-penalized Cox workflow was used for predictor selection, followed by unpenalized Cox refitting. Model performance was evaluated using C-statistics, time-specific Brier scores, calibration, bootstrap internal validation, and 5-year time-dependent decision curve analysis.
Results: During a median follow-up of 61 months, 126 first recurrence events were observed (88 training and 38 validation). The final combined model retained RCB category, triple-negative breast cancer (TNBC), CD8 status, and CD163-positive macrophage proportion. The combined model achieved C-statistics of 0.79 (95% CI 0.75-0.83) in the training cohort and 0.75 (95% CI 0.69-0.81) in the internal validation cohort, with corresponding 5-year Brier scores of 0.142 and 0.158. Bootstrap internal validation yielded an optimism-corrected C-statistic of 0.772 and calibration slope of 0.92. The combined model provided the greatest net benefit among the evaluated models over 5-year threshold probabilities of approximately 15%-45%.
Conclusions: Integration of RCB with TNBC status and post-neoadjuvant CD8 and CD163-positive macrophage features may provide incremental prognostic information for recurrence after neoadjuvant therapy. The model showed acceptable internal discrimination and calibration, but external validation is required before clinical implementation or treatment-directed use.

Keywords:  breast cancer; immune microenvironment; neoadjuvant therapy; residual cancer burden; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3389/fonc.2026.1925326
Cancer Discov. 2026 Oct 26.

hnRNPA2B1 sustains antitumor T cell immunity and enhances cancer immunotherapy through IDR-dependent stabilization of chromatin-regulatory transcripts.

Shi Liu, Wenyan Gao, Menghan Hu, Lingyun Long, Hongyan Chen, Yun Zhang, Xuetao Cao, Zhihua Liu.

T cell exhaustion within the tumor microenvironment poses a major barrier to effective cancer immunotherapy, yet the epigenetic mechanisms that preserve functional states of tumor-infiltrating T cells remain incompletely understood. Here, using an in vivo CRISPR screen, we identify heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2B1) as a regulator of CD8+ T cell fitness. Transcriptomic analyses of cancer patient cohorts reveal that hnRNPA2B1 is preferentially enriched in a proliferative, intermediate exhausted T cell subset associated with favorable clinical outcomes. Genetic ablation of hnRNPA2B1 in CD8+ T cells accelerates terminal exhaustion, impairs antitumor immunity, and diminishes responsiveness to immune checkpoint blockade. Mechanistically, hnRNPA2B1 forms nuclear puncta and stabilizes RNAs involved in chromatin organization, thereby maintaining chromatin accessibility that restrains exhaustion-associated programs while preserving proliferative and cytotoxic potential. Together, these findings establish hnRNPA2B1 as an RNA-mediated epigenetic regulator sustaining functional exhausted T cell states in tumors.

DOI: https://doi.org/10.1158/2159-8290.CD-26-0255
Methods Mol Biol. 2027 ;3089 375-386

Methods for Analyzing Immune Infiltrates and Spatial Organization Within the Melanoma Microenvironment Using Multiplex Immunofluorescence.

Jackson R Clark, Kelly T Smith, Marya Dunlap-Brown, Timothy N J Bullock, Craig L Slingluff, Ileana S Mauldin.

  Immunotherapies improve survival in patients with melanoma by enhancing the immune system's ability to recognize and eliminate tumor cells. Understanding the immunological effects of these therapies requires evaluation of both changes in lymphocyte composition and activation within the tumor microenvironment (TME) as well as the spatial organization of these cells. Increased lymphocyte infiltration and distinct patterns of immune cell distribution within the TME have been associated with improved survival and enhanced response to immunotherapy in melanoma. However, relatively few methodologies currently enable detailed characterization of both the structural organization and immunological features of tumor-infiltrating lymphocytes. In this chapter, we describe multiplex immunofluorescence staining approaches for the identification, characterization, and quantification of tumor-infiltrating lymphocytes in melanoma.

Keywords:  IHC; Immune infiltrates; Melanoma; Multiplex immunofluorescence histology; Spatial organization

DOI:  https://doi.org/10.1007/978-1-0716-5619-8_26
Cancers (Basel). 2026 Oct 15. pii: 3325. [Epub ahead of print]18(20):

Reprogramming the Melanoma Tumor Microenvironment: From Cellular Crosstalk to Cellular Immunotherapy.

Enar Jumaniyazova, Elena Gantsova, Genrikh Kazarian, Irina Charyeva, Timur Fatkhudinov.

  Melanoma is the most aggressive and rapidly metastasizing malignant neoplasm of the skin and mucous membranes, arising from melanocytes of neuroectodermal origin. Despite substantial advances in immune checkpoint inhibitors and targeted therapy, a significant proportion of patients develop primary or acquired resistance, which is largely attributable to the complex interactions between tumor cells and the tumor microenvironment (TME). This review presents a comprehensive analysis of the cellular and molecular components of the melanoma TME and their contribution to immunosuppression, tumor progression, and therapeutic resistance. The role of keratinocytes as the primary niche-forming cells in the epidermis is examined, including their reprogramming during melanomagenesis and their involvement in drug resistance. Particular attention is given to cancer-associated fibroblasts (CAFs), which orchestrate stromal remodeling, promote immune exclusion, and limit the efficacy of immunotherapy. The phenotypic and functional heterogeneity of innate and adaptive immune cells-macrophages, granulocytes, natural killer cells, dendritic cells, T and B lymphocytes-is discussed in the context of their dual role in antitumor immunity and immune escape. A detailed analysis of immune editing mechanisms, including defective antigen recognition, resistance to apoptosis, and the establishment of an immunosuppressive microenvironment, is provided. The review further summarizes the current landscape of cellular immunotherapy for melanoma. Clinically established approaches, including tumor-infiltrating lymphocytes (TILs) and CAR-T cells, are discussed with reference to completed phase II-III clinical trials, whereas emerging strategies such as CAR-macrophages and dendritic cell vaccines are presented as largely preclinical or early-phase investigational platforms. It is concluded that the melanoma TME functions as a unified immunosuppressive network rather than a collection of individual cell populations. Based on currently available comparative clinical evidence, reprogramming this network through the combination of cellular platforms with immune checkpoint inhibitors, stromal modulators, and macrophage-repolarizing agents, supported by personalized assessment of TME composition, represents a rational and actively investigated therapeutic direction.

Keywords:  cellular immune therapy; immune cells; immunotherapy; melanoma; tumor microenvironment; tumor-associated macrophages; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3390/cancers18203325
bioRxiv. 2026 Sep 01. pii: 2026.07.23.740424. [Epub ahead of print]

The Y chromosome gene KDM5D restrains CD8+ T cell antitumor immunity through TCR and cholesterol-exhaustion programs.

Jiexi Li, Chi Yin Ching, Aviad Ben-Shmuel, Paulino Tallón de Lara, Jingjing Liu, Jiang Shan, Chaohao Li, Ziyuan Zhang, William Huaiyang Wu, Max Slotnik, Xiaotian Wang, Raul Caballero Montes, Abhinav K Jain, Nicholas Hornstein, Fadl Zeineddine, Mohammad Zeineddine, Scott Eric Woodman, Natividad Roberto Fuentes, Denise J Spring, John Paul Shen, Scott Kopetz, Ronald A DePinho.

Sex differences in immunity shape cancer risk, autoimmunity, and responses to immunotherapy, yet the sex-chromosome genes that regulate antitumor T cell function remain incompletely defined. Here, we identify the Y chromosome-encoded KDM5D histone demethylase as a male-specific suppressor of CD8+ T cell antitumor immunity. In murine colorectal cancer (CRC) models, male CD8+ T cells displayed reduced cytokine production, proliferation, cytotoxicity, TCRβ abundance, and proximal TCR signaling relative to female CD8+ T cells. CRISPR-RNP-mediated KDM5D depletion in male CD8+ T cells enhanced effector function, increased TCRβ expression, augmented TCR signaling, and improved tumor control after adoptive transfer. Transcriptomic and functional analyses further linked KDM5D to cholesterol biosynthesis and exhaustion-associated programs, with KDM5D depletion reducing SREBP2/XBP1-associated cholesterol and exhaustion signatures. Correspondingly, human CRC single-cell analyses supported the clinical relevance of this axis, showing enrichment of exhausted and cholesterol-associated CD8+ T cell states in male tumors. Pharmacologic inhibition of cholesterol biosynthesis with lovastatin partially attenuated select exhaustion-associated markers in male CD8+ T cells and delayed tumor growth in vivo. Together, these findings define KDM5D as a sex chromosome-encoded regulator of male CD8+ T cell dysfunction and point to cholesterol-exhaustion programs as a potential therapeutic vulnerability in male CRC.

DOI: https://doi.org/10.64898/2026.07.23.740424
Anticancer Res. 2026 Nov;46(11): 6099-6111

Tumor Immune Microenvironment and VAF-inferred BRCA1/2 Loss of Heterozygosity in Japanese Patients With Germline BRCA1/2-mutated Breast Cancer.

Yukino Kawamura, Kotaro Mori, Fumihiko Takeuchi, Junko Kawano, Yasuaki Sagara, Norihiro Kato, Yasuyo Ohi, Akira I Hida, Chikako Shimizu, Akihiko Shimomura.

   BACKGROUND/AIM: Germline BRCA1/2 mutations (gBRCA1/2m) confer a markedly increased lifetime risk of breast cancer. Although diverse second-hit mechanisms lead to biallelic inactivation of BRCA1/2, their relationship with the immune microenvironment remains poorly understood, particularly in Japanese populations.
PATIENTS AND METHODS: We retrospectively analyzed 22 Japanese patients with gBRCA1/2m breast cancer who underwent curative surgery. Pathological evaluation included assessment of tumor-infiltrating lymphocytes (TILs), CD4+, CD8+, and FOXP3+ T-cell infiltration, and PD-L1 expression. Targeted next-generation sequencing of tumor samples identified somatic alterations. Loss of heterozygosity (LOH) at the BRCA1/2 locus was assessed using tumor-only sequencing data based on variant allele frequency (VAF). After adjustment for tumor cellularity, LOH was stringently defined as a modified VAF ≥0.90; cases not meeting this threshold were classified as non-LOH.
RESULTS: Twelve patients carried gBRCA1m and 10 carried gBRCA2m. High immune infiltration and frequent PD-L1 expression (68.2%) were observed, with a trend toward greater immune activity in BRCA1-mutated tumors. Of 14 cases with reportable sequencing results, LOH status was evaluable in 13. LOH was identified in 6 (46.2%) cases. LOH was more frequent in BRCA2-mutated tumors (75.0%) than in BRCA1-mutated tumors (33.3%). No differences were observed between LOH and non-LOH groups in CD4+, FOXP3+, or CD8+ T-cell infiltration, stromal TIL levels, PD-L1 expression, or disease-free survival.
CONCLUSION: In this Japanese cohort of BRCA-mutated breast cancer, integrated genomic and pathological analyses revealed heterogeneity in second-hit mechanisms and immune microenvironment features and suggested that detectable LOH does not fully account for immune activation in these tumors.

Keywords:  BRCA1; BRCA2; Breast cancer; germline mutations; retrospective analysis; second-hit patterns; tumor microenvironment

DOI:  https://doi.org/10.21873/anticanres.18441
bioRxiv. 2026 Oct 07. pii: 2026.10.05.756899. [Epub ahead of print]

Programmable antigen-presenting scaffolds expand rare tumor-antigen-specific T cells from peripheral blood.

Jessica DeVilla, Yoav Binenbaum, Nicoletta Cieri, David Zhang, Helen Ji, Vittoria Martinolli D'Arcy, Giacomo Oliveira, Michael Manos, Donna Neuberg, Jerome Ritz, Patrick A Ott, David Mooney, Catherine J Wu.

Adoptive cellular therapies can be effective against solid tumors but rely on the identification and expansion of rare tumor-reactive T cells (TRTs). Here, we utilize antigen-presenting cell-mimetic scaffolds (APC-ms), a modular biomaterial platform that presents defined peptide-HLA and co-stimulatory ligands on a fluid lipid bilayer to selectively recover rare antigen-specific T cells from peripheral blood. Independent variation of peptide-HLA, co-stimulatory ligands, and cytokine support defined a formulation that enriched MART-1-specific T cells to above 2% of CD8 + T cells in 35 of 50 HLA-A*02:01 + healthy donors from low or undetectable baseline frequencies. Compared with mo-DCs, APC-ms achieved greater fold expansion of MART-1-specific T cells with increased functional avidity and preservation of an early differentiation phenotype, and generated a higher number of MART-1-specific clonotypes. In five of eight patients with treatment-naïve melanoma, APC-ms could enrich MART-1-specific T cells to a median of 13.7% of CD8 + cells (range 0.30 - 57.2%) Moreover, APC-ms could expand T cells specific for an HLA-A*24:01-restricted personal neoantigen that were previously undetectable in peripheral blood of a patient with melanoma. These findings establish APC-ms as a tunable platform for the recovery of rare TRTs from peripheral blood, supporting its further development for therapeutic TCR discovery and personalized adoptive cell therapy.

DOI: https://doi.org/10.64898/2026.10.05.756899
Methods Mol Biol. 2027 ;3089 591-601

Generation of Murine Chimeric Antigen Receptor T Cells for Adoptive T Cell Therapy for Melanoma.

Amorette Barber.

  Expression of chimeric antigen receptors enables T cells to be redirected toward melanoma-associated antigens in an MHC-independent manner. Retroviral transduction can be used to introduce these receptor constructs into murine CD4⁺ and CD8⁺ T cells. Here, we outline the generation of retroviral supernatants and the activation, transduction, expansion, and selection of murine T cells expressing the chimeric-PD1 receptor. This protocol can be adapted for any murine chimeric antigen receptor construct.

Keywords:  Chimeric antigen receptor; Immunotherapy; Melanoma; Mouse; Retroviral transduction; T cell

DOI:  https://doi.org/10.1007/978-1-0716-5619-8_39
Adv Mater. 2026 Oct 26. pii: e75515. [Epub ahead of print]

Time-Encoded Ligand Presentation via a Magnetically Actuated Hydrogel Mitigates T Cell Exhaustion.

Xuemei Wang, Runyao Wang, Guangli Xiang, Xueyou Zhang, Luyang Wang, Wai Ki Wong, Yingrui Deng, Yongkang Yang, Jiwon Oh, Xian Xie, Zhinan Yang, Nathanael Chun Him Lai, Bohan Yin, Kunyu Zhang, Liming Bian, Siu Hong Dexter Wong.

  Adoptive T cell therapy (ACT) requires ex vivo expansion of tumor-reactive CD8+ T cells, yet prolonged antigenic stimulation frequently induces exhaustion, limiting persistence and therapeutic efficacy. Here, we present a magnetically actuated, extracellular matrix-mimetic hydrogel that enables reversible control of T cell receptor (TCR) stimulation timing. By embedding αCD3ε/αCD28-functionalized magnetic nanoparticles within a soft hyaluronic acid matrix, external magnetic fields dynamically expose or conceal stimulatory ligands, allowing programmable alternation between activation and rest. Continuous ligand presentation sustains TCR signaling and drives inhibitory receptor expression and functional exhaustion. In contrast, alternating activation-rest cycles periodically attenuate proximal TCR signaling, reduce exhaustion-associated marker expression, and bias differentiation toward central memory-like states while preserving cytokine production and recall capacity. T cells expanded under intermittent stimulation exhibit enhanced antigen-specific cytotoxicity, reduced dependence on PD-1 blockade, and superior tumor control in vivo, together with increased CD8+ T cell abundance across analyzed tissues. These results identify temporal control of mechanochemical signaling as a materials-encoded regulator of T cell fate and provide a non-genetic strategy for generating functionally competent T cell products with reduced exhaustion-associated dysfunction and improved antitumor activity.

Keywords:  T cell exhaustion; adoptive cell therapy; dynamic ligand presentation; immuno‐materials engineering; magnetically actuated hydrogel; time‐encoded signaling

DOI:  https://doi.org/10.1002/adma.75515
Biomedicines. 2026 Oct 19. pii: 2390. [Epub ahead of print]14(10):

Antibody Targeting Soluble NKG2D Ligand MIC Enhances Adoptive T-Cell Therapy in an NK Cell-Dependent Mechanism.

Fahmin Basher, Tyler A Smith, Mark P Rubinstein, Jennifer D Wu.

  Background/Objectives: Antigen-specific adoptive CD8+ T-cell therapy has demonstrated promising clinical activity in selected cancers; however, its broader efficacy against solid tumors remains limited by the immunosuppressive tumor microenvironment (TME), which compromises the persistence and function of transferred T-cells. A clinically relevant component of the human solid-tumor TME is soluble MHC class I chain-related molecules A and B (sMICA and sMICB, collectively termed sMIC), which are tumor-derived soluble NKG2D ligands that are not naturally expressed in mice. sMIC promotes tumor immune evasion through multiple mechanisms, but whether therapeutic clearance of sMIC can enhance the efficacy of adoptive T-cell therapy remains unclear. Methods: B16F10 melanoma cells engineered to express soluble MICB (B16F10-sMICB) were implanted subcutaneously into syngeneic B6/MICB mice. Once tumors became palpable, mice were randomized to four treatment groups: (1) no treatment; (2) adoptive transfer of gp100-specific Pmel CD8+T cells; (3) the sMIC-targeting monoclonal antibody B10G5; or (4) B10G5 combined with Pmel adoptive cell transfer. Pmel CD8+T cells were administered once intravenously, and B10G5 was administered intraperitoneally twice weekly at approximately 3 mg/kg. Tumor growth and body weight were monitored twice weekly. To determine the contribution of natural killer (NK) cells to the therapeutic effect of the combination treatment, NK cells were depleted using the anti-NK1.1 antibody PK136. The impact of NK cells on CD8+T cell function during B10G5/sMIC stimulation was further evaluated ex vivo. Results: Therapeutic targeting of sMIC with B10G5 significantly enhanced the antitumor efficacy of adoptively transferred Pmel CD8+T cells and was associated with increased donor T-cell persistence, tumor infiltration, and antigen-specific effector function. NK cell depletion markedly diminished the antitumor efficacy of the B10G5/Pmel combination and reduced the abundance, activation, and effector function of donor CD8+ T-cells. Consistent with these in vivo findings, ex vivo depletion of NK cells impaired IFN-γ production by NKG2D+CD8+ T-cells during B10G5/sMIC stimulation, supporting a functional role for NK cells in sustaining CD8+T cell responses. Conclusions: These findings provide evidence that therapeutic targeting of tumor-derived sMIC can enhance the efficacy of antigen-specific adoptive CD8+T cell therapy by promoting the persistence and function of transferred T-cells. Importantly, our results identify NK cells as critical contributors to the enhanced therapeutic response, revealing functional cooperation between NK cells and adoptively transferred CD8+T cells during sMIC-targeted therapy. Together, these findings provide preclinical proof-of-concept for combining sMIC-targeting antibodies with adoptive T-cell therapy to improve antitumor immunity against sMIC+ solid tumors.

Keywords:  NK cells; NKG2D; adoptive T cell therapy; antibody; soluble MIC (sMIC)

DOI:  https://doi.org/10.3390/biomedicines14102390
Diseases. 2026 Oct 03. pii: 367. [Epub ahead of print]14(10):

Immune- and Stromal-Associated Tissue-Context Profiles in Pretreatment Triple-Negative Breast Cancer: An Exploratory Institutional Cohort Study.

Valeria Zuccalà, Nicoletta Staropoli, Caterina Labanca, Gabriele Ricciardi, Pietro Tralongo, Vincenzo Fiorentino, Elisabetta Procopio, Maria Renne, Antonio Ieni, Maurizio Martini, Pierfrancesco Tassone, Pierosandro Tagliaferri.

   BACKGROUND/OBJECTIVES: We examined combinations of immune- and stromal-associated features in pretreatment triple-negative breast cancer (TNBC) biopsies and explored their clinical and germline associations.
METHODS: This retrospective study included 24 women with nonmetastatic TNBC from a clinically selected BRCA1/BRCA2-tested cohort. Two pathologists, blinded to clinical and germline data, jointly assessed stromal tumor-infiltrating lymphocytes (sTILs), programmed death-ligand 1 (PD-L1), CD68/CD163-positive macrophages, and CD117/tryptase-positive mast cells. Profiles were assigned before unblinding. A second blinded joint sTIL assessment provided continuous estimates for subsequent hierarchical clustering; profile assignments remained unchanged.
RESULTS: Profiles I-III comprised seven, 13, and four tumors, respectively. Profile I combined higher sTILs and PD-L1 with a CD68-high/CD163-low pattern and rare mast cells. Profile II showed lower sTILs and PD-L1, a CD68-low/CD163-high pattern, and dense mast-cell hotspots; Profile III had minimal sTILs, low macrophage counts, and no mast cells detected in evaluated fields. Clustering reproduced these assignments. All four patients with pathogenic/likely pathogenic germline BRCA1/BRCA2 variants were in Profile I (adjusted p = 0.030); omitting one positive case at a time increased the adjusted p value to 0.068. Seven of 16 neoadjuvant-treated patients achieved pathologic complete response. Five posttreatment recurrences and seven deaths were recorded; exploratory comparisons of pathologic complete response and overall survival were inconclusive.
CONCLUSIONS: These profiles describe different combinations of lymphocytic infiltration, PD-L1 expression, macrophage counts, and mast-cell hotspots. The profiles and the exploratory BRCA1/BRCA2 finding require validation in larger, independent cohorts before clinical use.

Keywords:  PD-L1; androgen receptor; germline BRCA1/BRCA2; hierarchical clustering; mast cells; stromal tumor-infiltrating lymphocytes; tissue-context profile; triple-negative breast cancer; tumor-associated macrophages

DOI:  https://doi.org/10.3390/diseases14100367
Clin Ter. 2026 Nov-Dec;177(6):177(6): 1474-1482

Clinicopathological, Molecular, and Immune Profiling in Colorectal Cancer: Associations Between KRAS/BRAF Alterations, dMMR/MSI-H Status, and Tumor-Infiltrating Lymphocytes: A Retrospective Cross-Sectional Study.

Nguyen Quoc Dat, Tran Ngoc Minh, Nguyen Van Chu, Nguyen Sy Lanh, Ha Xuan Hop, Bui Tien Cong.

   Background: To characterize the molecular and immune profiles of resected colorectal cancer (CRC), including KRAS/NRAS/BRAF mutations, deficient mismatch repair/high microsatellite instability (dMMR/MSI-H), and tumor-infiltrating lymphocyte (TIL) positivity, using routinely available pathological methods, and to identify independent clinicopathological and molecular factors associated with TIL positivity and dMMR/MSI-H status.
Methods: This retrospective cross-sectional study included 272 patients with primary colorectal adenocarcinoma who underwent surgical resection between 2018 and 2024. KRAS, NRAS, and BRAF mutations were analyzed by real-time polymerase chain reaction. Mismatch repair protein expression was evaluated by immunohistochemistry (IHC), and tumors with loss of at least one mismatch repair protein were classified as dMMR using IHC as a surrogate method for MSI-H status. TIL were assessed on hematoxylin and eosin (H&E)-stained sections.
Results: KRAS mutations were detected in 40.8% of tumors, while BRAF and NRAS mutations were identified in 4.0% and 2.9% of cases, respectively. dMMR/MSI-H was present in 9.6% of tumors. Positive TIL status was observed in 35.3% of cases. Univariable analysis showed that dMMR/MSI-H status and fungating tumor morphology were significantly associated with higher TIL positivity, although these associations became borderline significant after multivariable adjustment. Independent factors associated with dMMR/MSI-H status included younger age (adjusted OR = 0.29, 95% CI: 0.12-0.69), proximal tumor location (adjusted OR = 0.25, 95% CI: 0.10-0.64), and poor tumor differentiation (adjusted OR = 2.80, 95% CI: 1.16-6.76). Subgroup analysis also showed that dMMR-related groups were more likely to have tumors in the proximal colon, poor differentiation, younger age, and higher lymphocytic infiltration.
Conclusions: Integrated clinicopathological and molecular analysis identified specific CRC features associated with dMMR/MSI-H status, including proximal tumor location, poor differentiation, younger patient age, and a trend toward increased lymphocytic infiltration. Borderline associations were also observed between TIL positivity, fungating tumor morphology, and dMMR/MSI-H phenotypes. These findings support the feasibility of combined molecular and immune evaluation using routinely available pathological methods, particularly in resource-limited settings.

Keywords:  Colorectal cancer; KRAS; BRAF; mismatch repair deficiency; microsatellite instability; tumor-infiltrating lymphocytes; molecular pathology; cross-sectional study

DOI:  https://doi.org/10.7417/CT.2026.2153
J Immunol. 2026 Oct 01. pii: vkag262. [Epub ahead of print]215(10):

Experimental high blood glucose metabolically rejuvenates CD8+ T cells for improved control of tumor growth through an IGF1-IGF1R axis.

Anirban Sarkar, Sukanya Dhar, Saurav Bera, Mohona Chakravarti, Ayushi Verma, Muqtada Ali Khan, Parash Prasad, Anupam Das, Pritha Roy Choudhury, Jasmine Sultana, Juhina Das, Akata Saha, Avishek Bhuniya, Ipsita Guha, Shayani Dasgupta, Sib Sankar Roy, Saptak Banerjee, Subir Roy, Debarati Bhar, Walter J Storkus, Rathindranath Baral, Dipak Datta, Anamika Bose.

  Epidemiologic studies suggest that patients with pre-existing type 1 diabetes have a decreased risk of developing melanoma, prostate cancer, and breast cancer, although the underlying mechanism remains unresolved. In translational model studies, we observed that a streptozotocin-induced high blood glucose model exhibited restricted melanoma and carcinoma (mammary, lung, and colon) growth in association with extended overall survival. Tumor-infiltrating CD8+ T cells were responsible for tumor growth restriction. Tumor-infiltrating CD8+ T cells but not tumor cells themselves exhibited higher glycolytic and cytotoxic activity in high-glucose hosts. Such improved antitumor T cell function was linked to upregulated expression of Igf1, Igf1R, and phospho-mTOR in CD8+ T cells in the tumor microenvironment. High blood glucose patient-derived peripheral CD8+ T cells displayed superior activation in vitro after allogenic tumor antigen stimulation versus nondiabetic CD8+ T cells. Activation of high-glucose-patient CD8+ T cells was sensitive to targeted antagonism of IGF1R and mTOR, supporting the operational involvement of the IGF1R-mTOR signaling axis. Our results suggest that selective activation of the intrinsic IGF1R-mTOR signaling axis in CD8+ T cells serves as a preferred endpoint to achieving more effective immunotherapy outcomes in cancer patients.

Keywords:  CD8 T cells; high glucose model; insulin like growth factor 1; insulin like growth factor 1 receptor; tumor microenvironment

DOI:  https://doi.org/10.1093/jimmun/vkag262
Life Sci. 2026 Oct 24. pii: S0024-3205(26)00557-6. [Epub ahead of print]406 124748

Overcoming the efficacy-safety dilemma of cytokine therapy for tissue-resident memory T cells in cancer immunotherapy: Intrinsic sensitization and extrinsic optimization.

Yao Xiao, Zi-Zhan Li, Zhi-Jun Sun.

  Tissue-resident memory T (TRM) cells are important contributors to immune checkpoint blockade therapy response in several cancers, yet their therapeutic potential remains underutilized. The cytokine-mediated enhancement of TRM cells represents a promising strategy to amplify antitumor immunity. However, clinical translation is hindered by an efficacy-safety dilemma. Potent cytokines such as IL-12 and IL-15 induce severe systemic toxicity; other potent cytokines such as TGF-β and IL-1 carry inherent immunosuppressive risks. To overcome this barrier, a dual-strategy framework is proposed. Intrinsic sensitization involves the use of multidimensional regulatory networks such as those involving transcription factors, metabolic pathways, hypoxia signaling, and host aging to enhance TRM responsiveness to cytokines rather than increasing the dose. Complementing this, extrinsic optimization minimizes risks through targeted delivery systems, tumor microenvironment (TME) modulation for endogenous release, rational cytokine combination, and protein engineering coupled with rational selection. Collectively, this roadmap outlines how to harness cytokine networks safely and effectively, enabling precision TRM-based antitumor immunotherapy.

Keywords:  Cancer immunotherapy; Cytokine therapy; Efficacy–safety dilemma; Extrinsic optimization; Intrinsic sensitization; Tissue-resident memory T cells

DOI:  https://doi.org/10.1016/j.lfs.2026.124748
Methods Mol Biol. 2027 ;3089 363-374

A Method for Quantifying Immune Infiltrates and Tertiary Lymphoid Structures in Murine Melanoma Tumors by Multiplex Immunofluorescence.

Jackson R Clark, Pallavi Rustogi, Ileana S Mauldin.

  The presence of tertiary lymphoid structures (TLS) and elevated levels of tumor-infiltrating lymphocytes have been linked to improved survival outcomes in several solid cancers, including melanoma. Despite this association, the precise function and immunological impact of TLS remain incompletely understood, as do factors that drive their formation and which might potentially be leveraged to enhance patient outcomes. Functional studies using murine models may help clarify the biological roles of TLS; however, relatively few methodologies currently exist that allow for detailed assessment of the structural organization and functional characteristics of TLS within solid tumors. To advance understanding of the biology underlying TLS, this chapter describes multiplex immunohistochemistry panels designed for the identification, characterization, and quantification of TLS and intratumoral immune infiltrates in melanoma. Although these methods are presented in the context of melanoma, they are broadly applicable and can be used to investigate TLS across a wide range of human cancers.

Keywords:  High endothelial venules; IHC; Immune infiltrates; Immunology; Melanoma; Multiplex immunofluorescence histology; Tertiary lymphoid structures

DOI:  https://doi.org/10.1007/978-1-0716-5619-8_25
J Radiat Res. 2026 Oct 27. pii: rrag089. [Epub ahead of print]

Association of pretreatment immune microenvironment profile with boron neutron capture therapy efficacy in cutaneous tumors: results from an ancillary multiplex immunofluorescence study.

Masahiro Ueno, Tairo Kashihara, Satoshi Nakamura, Hana Endo, Taisuke Mori, Satsuki Kishikawa, Eiji Nakano, Dai Ogata, Kenjiro Namikawa, Naoya Yamazaki, Ryuta Saito, Hiroshi Igaki.

  Boron neutron capture therapy (BNCT), an emerging novel radiotherapy modality, has shown immunological relevance in preclinical models. Although the safety and efficacy of BNCT for cutaneous tumors have been evaluated in a Phase I clinical trial, studies using human clinical specimens remain limited. In this exploratory translational study, we aimed to investigate pretreatment immune microenvironmental features in patients treated with BNCT and to descriptively assess their relationship with clinical response. To this end, formalin-fixed paraffin-embedded pretreatment biopsy samples obtained from a Phase I clinical trial of BNCT for cutaneous tumors were analyzed in three responders and three non-responders. Multiplex immunofluorescent staining was performed using CD8+ T-cell and dendritic cell (DC) antibody panels to assess the phenotypes and spatial relationships of immune cells within the tumor microenvironment. At baseline, responders tended to show higher densities of total T cells, CD8+ T cells, DCs and mature DC-like cells than non-responders. CD8+ T cells in responders tended to include higher proportions of granzyme B+ and programmed cell death protein 1-positive cells, and spatial analysis showed closer proximity between CD8+ T cells and mature DC-like cells. In contrast, the proportions of programmed death-ligand 1-positive cells among DCs and tumor cells were higher in non-responders than in responders. These descriptive findings suggest that pretreatment immune microenvironmental features may warrant further investigation as potential factors related to clinical response to BNCT.

Keywords:  T-lymphocytes; boron neutron capture therapy; programmed cell death 1 ligand 1 protein; programmed cell death 1 receptor; radiotherapy; tumor microenvironment

DOI:  https://doi.org/10.1093/jrr/rrag089