bims-tuinly 2026-07-05 papers

bims-tuinly

Biomed News

on Tumor-infiltrating lymphocytes therapy

Issue of 2026–07–05
28 papers selected by
Pierpaolo Ginefra, Ludwig Institute for Cancer Research

HLA-G expression in non-small cell lung cancer: prognostic significance and interplay with PD-L1 and CD8+ tumor-infiltrating lymphocytes.
AI-assisted image analysis of tumor-infiltrating lymphocytes as a prognostic marker in chemotherapy-naïve luminal breast cancer.
The atypical IκB factor IκBδ enhances CD8 T cell accumulation and effector functions in solid tumors.
Antigen-scaffolds loaded with hyper-stable neoleukin-2/15 expand antigen-specific T cells with a favorable phenotype for adoptive cell therapy.
Tailored HER2 ECD mRNA-LNP vaccines boost CD8+ T cell-mediated antitumor immunity for HER2-positive tumor suppression.
Pretreatment Tumor-infiltrating CD8+ T Cell Numbers Are Associated With the Response to Tremelimumab Plus Durvalumab in Hepatocellular Carcinoma.
Longitudinal evaluation of tumor-infiltrating lymphocyte scoring using automated region of interest registration in breast cancer.
Prognostic value of an integrated immune-inflammatory phenotype in surgically treated cervical cancer: survival modeling and immunohistochemical validation.
Circulating and brain-resident memory CD8+ T cells seed distinct bystander T RM -like populations in glioblastoma.
Machine learning-driven identification and immunohistochemical validation of an integrated immune-inflammatory phenotype for disease-free survival stratification in breast cancer.
A pragmatic workflow for advanced NSCLC diagnosis in routine practice: Integrating histopathology, PD-L1 scoring, and EGFR testing.
Dominant T cell receptor clonotypes in adrenocorticotropic hormone-secreting pituitary carcinoma are the highest-frequency clones among CD4+ and CD8+ cells in peripheral blood during effective anti-PD-1 therapy.
Engineered immune cells and extracellular vesicles target tumour microenvironment barriers in solid tumour immunotherapy.
Homologous recombination deficiency correlates with tumor-infiltrating lymphocytes and predicts patient outcomes in hormone receptor-positive/HER2-negative breast cancer.
Fusion protein-mediated costimulation in engineered T cells: from intrinsic signaling to tumor microenvironment rewiring.
PARP inhibition combined with a T-cell receptor β chain-directed antibody fusion molecule drives polyclonal antitumor immunity and tumor regression.
Interleukin 12-tethered nano-aluminum adjuvant orchestrates environmental immunomodulation to empower cytotoxic T cells against solid tumors.
Single-cell transcriptomic analysis reveals tumor-immune determinants of lymph node colonization and progression in thyroid cancer.
Advances and prospects in cell therapy for cancer: explorations from T cells to stem cells.
A randomized phase 2 study of combination atezolizumab and varlilumab (CDX-1127) with or without cobimetinib in previously-treated unresectable biliary tract cancer.
Sustained A2AR expression and loss paradoxically promote CD8+ T cell exhaustion.
Single-cell transcriptomic analysis identifies a tumor-enriched GIMAP7⁺ CD4⁺ naïve T cell population with a unique immunoregulatory state in lung adenocarcinoma.
CD47 monoclonal antibody enhances the inhibitory effect of anti-HER2 chimeric antigen receptor macrophages on ovarian cancer.
R-5780, a SagA-engineered Lactococcus lactis, is a safe oral synthetic-biology microbial therapy that potentiates PD-1 blockade.
Editorial: Exploring unconventional T cells and CAR-T-cells in cancer immunotherapy.
NKG2C + CD27 - Defines Human CD8 + Regulatory T Cells.
Programmed cell death ligand 1 in correlation with BRAFV600E mutation, molecular characteristics and biological behavior in ameloblastoma.
Opposing Dynamics of CD4 + Regulatory and CD8 + Foxp3+ T Cells Characterize Immune Imbalance in Rheumatoid Arthritis.

Front Immunol. 2026 ;17 1732852

HLA-G expression in non-small cell lung cancer: prognostic significance and interplay with PD-L1 and CD8+ tumor-infiltrating lymphocytes.

Giulia Querzoli, Giuseppe Bogina, Marcella Marconi, Nicola Tumino, Paola Vacca, Luisella Righi, Sara Pilotto, Anna Caliò, Luca Cima, Stefano Gobbo, Matthew J Cecchini, Gaetano Paolino, Francesco Ciompi, Roberto S Accolla, Aldo Scarpa, Gianluigi Lunardi, Emanuela Marcenaro, Lorenzo Moretta, Giuseppe Zamboni, Enrico Munari.

   Introduction: HLA-G is a non-classical major histocompatibility complex class I molecule with potent immunosuppressive activity and is increasingly recognized as an immune-checkpoint axis in cancer. Its prognostic significance in non-small cell lung cancer (NSCLC), particularly in relation to PD-L1 expression and CD8+ tumor-infiltrating lymphocytes (TILs), remains incompletely defined.
Methods: We retrospectively analyzed 314 surgically resected NSCLCs assembled in tissue microarrays and stained for HLA-G, PD-L1, and CD8. HLA-G and PD-L1 were scored as positive when ≥1% of tumor cells showed membranous staining, whereas CD8+ TIL density was digitally quantified and dichotomized using the cohort median (≥575 cells/mm²). Associations with clinicopathological variables and outcomes were assessed by Kaplan-Meier analysis and multivariable Cox regression.
Results: HLA-G was expressed in 50 of 314 tumors (16%), PD-L1 in 106 of 314 (33.8%), and high CD8 density in 160 of 314 (51%). In HLA-G-negative tumors, high CD8+ TIL density was associated with significantly prolonged disease-free survival (DFS) and overall survival (OS). In the overall cohort, the combined HLA-G-negative/CD8-high phenotype retained independent favorable prognostic significance for both DFS and OS. By contrast, CD8 density did not significantly stratify outcome in HLA-G-positive tumors, although these subgroup analyses were limited by small sample size.
Discussion: In combined biomarker analyses, the favorable prognostic effect of CD8+ TILs in PD-L1-negative tumors was maintained only when HLA-G was also absent. Within the PD-L1-positive/HLA-G-negative subgroup, high CD8 density was independently associated with improved DFS but not OS. Integrating HLA-G with PD-L1 and CD8 assessment may refine prognostic stratification and help identify patients who could benefit from HLA-G-targeted strategies, alone or in combination with PD-1/PD-L1 blockade.

Keywords:  HLA-G; PD-L1; immune evasion; non-small cell lung cancer; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3389/fimmu.2026.1732852
NPJ Breast Cancer. 2026 Jul 01.

AI-assisted image analysis of tumor-infiltrating lymphocytes as a prognostic marker in chemotherapy-naïve luminal breast cancer.

Dusan Rasic, Sandra Sinius Pouplier, Maj-Britt Jensen, Bent Ejlertsen, Roberto Salgado, Pekka Ruusuvuori, Johan Hartman, Mattias Rantalainen, Anne-Vibeke Lænkholm.

In triple-negative and HER2-positive breast cancer, tumor-infiltrating lymphocytes (TILs) are established predictive and prognostic biomarkers, but their role in luminal subtypes remains unclear. We investigated the prognostic significance of TILs using AI-based image analysis in 2298 luminal breast cancers from the DBCG99c cohort, classified by PAM50. Stromal (sTIL) and intraepithelial (iTIL) densities were quantified automatically using a commercial platform. Higher stromal TIL infiltration was associated with improved overall survival, particularly within the first 5 years (heterogeneity p = 0.01). In multivariable models, higher sTILs independently predicted lower risk of distant or any recurrence (sHR = 0.95, 95% CI 0.91-0.99) and improved survival (HR = 0.91, 95% CI 0.85-0.97 early; HR = 0.99, 95% CI 0.97-1.00 late). Intraepithelial TILs were not prognostic. No significant interactions were observed by molecular subtype, nodal status, or grade, although a nonsignificant trend toward stronger iTIL effects appeared in luminal B tumors. AI-based TIL quantification thus provides independent prognostic information in high-risk luminal breast cancer.

DOI: https://doi.org/10.1038/s41523-026-00999-w
bioRxiv. 2026 Jun 19. pii: 2026.06.17.732005. [Epub ahead of print]

The atypical IκB factor IκBδ enhances CD8 T cell accumulation and effector functions in solid tumors.

Barsha Dash, Xiaocui He, Arlet Lara-Custodio, Leo J Arteaga-Vazquez, Yue Zhao, Hannah Drum, Osamu Ikeda, Eric Johnson, Yuntian Zhu, Chen Zhang, Srikanth Battu, Edahí González-Avalos, Anjana Rao, Patrick G Hogan.

Two prominent mechanisms by which tumors fend off immune control are by constraining the ability of T cells and CAR T cells to survive and expand in the tumor, and by restraining their ability to sustain full cytotoxic capacity. We identified IκBδ, encoded by Nfkbid , a poorly characterized IκB family member, as a molecular lever that overcomes both of these constraints on anti-tumor CD8 + tumor-infiltrating lymphocytes (TILs). Nfkbid is an NFAT target gene that is expressed in CD8 + effector T cells and, at modest levels, in CD8 + TILs. We found that Nfkbid depletion impaired TIL accumulation, exacerbating the growth of solid tumors. On the other hand, ectopic IκBδ overexpression enhanced TIL expansion, reduced the expression of exhaustion-associated transcription factors and inhibitory receptors, and elevated cytotoxic molecule production, leading to enhanced tumor control. IκBδ has a shorter protein isoform that is identical in a core region spanning the ankyrin-repeat domain known to interact with NFκB proteins, but that lacks the ∼150-residue N-terminal region. We showed that the shared core region is sufficient to drive T cell accumulation, whereas the N-terminal peptide region is required for robust effector function and to counter exhaustion, underscoring that tumor-infiltrating CD8 + T cell accumulation and effector differentiation are separable programs. Our current study provides evidence that IκBδ, an atypical member of the NFκB family, is a lever to overcome two cardinal deficits that limit CD8 + TIL anti-tumor efficacy: impaired accumulation in the tumor and diminished effector function.

DOI: https://doi.org/10.64898/2026.06.17.732005
Front Immunol. 2026 ;17 1804678

Antigen-scaffolds loaded with hyper-stable neoleukin-2/15 expand antigen-specific T cells with a favorable phenotype for adoptive cell therapy.

Maria Ormhøj, Kamilla Kjærgaard Munk, Siri Tvingsholm, Keerthana Ramanathan, Amalie Kai Bentzen, Georgios Kladis, Gitte Nygaard Aasbjerg, Grigorii Nos, Tripti Tamhane, Hólmfridur Rósa Halldórsdóttir, Søren Nyboe Jakobsen, Marcus Svensson Frej, Kristoffer Haurum Johansen, Mohammad Kadivar, Sine Reker Hadrup.

   Introduction: Adoptive cell therapy (ACT) has shown promising results in cancer treatment, however, achieving effective ex vivo expansion of potent, functionally active, and cytotoxic T cells remains challenging. To address this challenge, we recently developed artificial antigen-presenting scaffolds (Ag-scaffolds) capable of expanding antigen-specific T cells with phenotypes favorable for ACT. Here, we compared the established technology using IL2/IL21-loaded Ag-scaffolds (Ag-IL2/21) with scaffolds incorporating Neoleukin-2/15 (Ag-Neo2/15), an engineered cytokine that selectively signals via IL-2Rβ/γ complexes to enhance CD8+ T cell proliferation while limiting regulatory T cell expansion.
Methods: Antigen-specific T cells were expanded ex vivo using Ag-IL2/21 or Ag-Neo2/15 scaffolds. Expansion efficiency, cytokine production, and cytotoxic activity were assessed. Functional and transcriptional states were profiled using single-cell sequencing, including cytotoxic and dysfunction gene signature scoring. T cell receptor (TCR) sequencing was performed to evaluate clonal expansion.
Results: Ag-Neo2/15 scaffolds supported robust expansion of antigen-specific CD8+ T cells at levels comparable to Ag-IL2/21 scaffolds. Ag-Neo2/15-expanded CD8+ T cells exhibited increased effector cytokine production and durable cytotoxicity against tumor cells. Single-cell transcriptomic analysis revealed higher cytotoxic gene signature scores, reduced dysfunctional scores, and increased expression of genes linked to increased proliferation in Ag-Neo2/15-expanded T cells. TCR sequencing demonstrated preferential expansion of CD8+ T cell clones enriched for transcriptional features associated with antigen-experienced effector states and sustained effector function.
Discussion: Ag-Neo2/15 scaffolds enhance the quality of ex vivo-expanded antigen-specific T cells by promoting a cytotoxic, less dysfunctional, and highly proliferative phenotype. These findings identify Ag-Neo2/15 as a promising improvement for ACT manufacturing, optimizing the balance between expansion capacity and durable effector function, with potential implications for improving cancer immunotherapy outcomes.

Keywords:  T-cell expansion; T-cells; adoptive cell therapy; artificial antigen-presenting scaffolds; neoleukin-2/15

DOI:  https://doi.org/10.3389/fimmu.2026.1804678
Drug Resist Updat. 2026 Jun 26. pii: S1368-7646(26)00090-7. [Epub ahead of print]88 101439

Tailored HER2 ECD mRNA-LNP vaccines boost CD8+ T cell-mediated antitumor immunity for HER2-positive tumor suppression.

Kexin Li, Zilin Ma, Xiaoya Li, Xiuping Chi, Ying Zhang, Cong Zhang, Wen Li, Mingyu Cui, Wei Qi, Yating Zhao, Jun Wang, Guogui Sun, Hongtao Zhang, Lianmei Zhao.

  HER2-positive tumors persist as a formidable therapeutic challenge despite advances in targeted agents, chemotherapy, and immunotherapy. Recently, messenger RNA (mRNA) vaccines have emerged as promising avenues for the treatment of advanced tumors and their recurrence, although their clinical application remains challenging. Herein, we developed lipid nanoparticle (LNP)-encapsulated nucleoside-modified mRNA vaccines encoding the modified extracellular domain (ECD) sequence from rat HER2 (rHER2 ECD mRNA-LNP) and human HER2 (hHER2 ECD mRNA-LNP). While both vaccines elicited effective tumor inhibition, the hHER2 ECD mRNA-LNP demonstrated superior prophylactic and therapeutic efficacy in vivo, and successfully induced potent, antigen-restricted cytotoxicity against human patient-derived organoids (PDOs) ex vivo. Mechanistically, it enhances both humoral and T cell responses, and reprograms CD8+ tumor-infiltrating lymphocytes (TILs) by reducing exhaustion, promoting effector maturation, and inducing memory T cells. Specifically, a reduction in naive-like T cell clusters coupled with an elevation in cycling T cell clusters underpins the vaccine's efficacy. Furthermore, HER2 epitope-specific T cell receptor β-chain variable/joining (Trbv/Trbj) combinations and complementarity-determining region 3 (CDR3) motifs were identified as key contributors to the antigen-specific tumoricidal activity. Crucially, combining the hHER2 ECD mRNA-LNP vaccine with programmed cell death-1 (PD-1) immune checkpoint blockade (ICB) achieves superior antitumor efficacy compared to vaccine monotherapy. Moreover, combining the hHER2 ECD mRNA-LNP vaccine with standard chemoradiotherapy effectively potentiates the therapeutic efficacy against HER2-positive tumors. These findings highlight a promising agent for oncology and suggest that modified HER2 ECD mRNA-LNPs represent a viable and highly effective therapeutic strategy against HER2-positive malignancies.

Keywords:  Breast cancer; HER2; Immunotherapy; Lipid nanoparticle; mRNA vaccine

DOI:  https://doi.org/10.1016/j.drup.2026.101439
Anticancer Res. 2026 Jul;46(7): 4077-4085

Pretreatment Tumor-infiltrating CD8+ T Cell Numbers Are Associated With the Response to Tremelimumab Plus Durvalumab in Hepatocellular Carcinoma.

Akifumi Kuwano, Masayoshi Yada, Kosuke Tanaka, Taikan Hamamoto, Kazuki Kurosaka, Hideo Suzuki, Yoshihiro Ohishi, Kenta Motomura.

   BACKGROUND/AIM: Hepatocellular carcinoma (HCC) shows heterogeneous responses to treatment with immune checkpoint inhibitors, and predictive biomarkers for dual immune checkpoint blockade remain insufficiently defined. The STRIDE regimen, combining tremelimumab and durvalumab, improves the survival of patients with unresectable HCC, but a substantial proportion of patients experience early progression. We investigated whether pretreatment intratumoral CD8+ tumor-infiltrating lymphocyte (TIL) density was associated with treatment response and progression-free survival (PFS) in patients treated with STRIDE.
PATIENTS AND METHODS: This retrospective, single-center study included 26 consecutive patients with unresectable HCC who received STRIDE and underwent pretreatment liver tumor biopsy. CD8+ TIL density was assessed using immunohistochemistry. Tumors were classified as CD8-high or CD8-low using a prespecified cutoff (15.9 cells/high-power field).
RESULTS: Fifteen patients were classified as CD8-high and 11 as CD8-low. The objective response rate was significantly higher in the CD8-high group than in the CD8-low group (60.0% vs. 9.1%, p=0.005). The median PFS was significantly longer in the CD8-high group [8.8 months, 95% confidence interval (CI)=3.0-24.5] than in the CD8-low group (2.0 months, 95%CI=0.8-7.2; p=0.019). The frequency of grade ≥2 immune-related adverse events did not differ significantly between groups.
CONCLUSION: Pretreatment intratumoral CD8+ TIL density was associated with an improved response and a longer PFS in STRIDE-treated HCC. Baseline tumor immune contexture may serve as a practical predictive biomarker for STRIDE. These findings are exploratory and hypothesis-generating, and require validation in larger prospective studies.

Keywords:  CD8+ T cells; Hepatocellular carcinoma; durvalumab; liver tumor biopsy; tremelimumab; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.21873/anticanres.18267
Breast Cancer Res. 2026 Jul 03.

Longitudinal evaluation of tumor-infiltrating lymphocyte scoring using automated region of interest registration in breast cancer.

Alessio Fiorin, Laia Adalid-Llansa, Laia Reverté, Esther Sauras-Colón, Noèlia Gallardo-Borràs, Hatem A Rashwan, Ramon Bosch-Príncep, Alba Fischer-Carles, Elena Goyda, Marylène Lejeune, Daniel Mata-Cano, Domènec Puig, Tábata Sánchez-Alcántara, Mikel Relloso Ortiz de Uriarte, Montserrat Llobera-Serentill, José Antonio Izuel-Navarro, Carlos López-Pablo.

   BACKGROUND: Tumor-infiltrating lymphocytes (TILs) are prognostic biomarkers in breast cancer (BC), particularly in HER2-positive and triple-negative subtypes. Assessment follows the international guidelines, in which pathologists evaluate whole hematoxylin and eosin (H&E)-stained slides while integrating representative regions of the invasive tumor. However, manual region selection can be labor-intensive, subjective, and may introduce variability, particularly across consecutive tissue sections. Automated region of interest (ROI) registration may mitigate this limitation, yet its impact on longitudinal TIL scoring has not been systematically evaluated. Here, we introduce three ROI registration strategies (direct, intermediate, and serial with/without quality control) and present an automated framework validated for consistent TIL scoring and clinical relevance in predicting relapse.
METHODS: We analyzed 104 invasive BC cases, each with 12 consecutive H&E slides. A pathologist annotated ROIs on both the first and twelfth slides. We registered these ROIs using the proposed strategies. We then evaluated them with performance metrics, including Intersection over Union (IoU), Dice Similarity Coefficient (DSC), failure rate, and execution time. Two pathologists scored TILs on manual and automated ROIs. We assessed longitudinal consistency between the first manual ROI and the twelfth slide's corresponding ROIs. We also tested whether the association between TIL score and patient relapse outcomes was preserved.
RESULTS: The direct registration strategy achieved the highest geometric accuracy (mean IoU = 0.650, DSC = 0.769), with < 1% failure rate and ∼5.8 s execution time. TIL scores for both manual and automated ROIs closely matched (P = 0.84), showing strong agreement (Spearman's ρ = 0.923, ICC = 0.936, CCC = 0.915, Cohen's κ = 0.786). Longitudinal analyses showed no significant variability across distant sections (P = 0.79 and P = 0.62). TIL concentrations differed significantly between patients with and without relapse, and this association was preserved on both the first and twelfth slides (P < 0.05).
CONCLUSIONS: We present the first automated framework for longitudinal ROI registration to support TIL scoring in BC. By reducing manual effort and variability, the framework supports scalable evaluation of immune biomarkers across tissue depth while preserving clinically relevant signals, supporting precision immuno-oncology applications.

Keywords:  Breast cancer; Histopathology; Image registration; Tumor-infiltrating lymphocytes; Whole slide images

DOI:  https://doi.org/10.1186/s13058-026-02333-5
Front Immunol. 2026 ;17 1850891

Prognostic value of an integrated immune-inflammatory phenotype in surgically treated cervical cancer: survival modeling and immunohistochemical validation.

Lin Ran, Zhaoan Lian, Yong Tian, Li Qin, Yingchun Xiang, Xiaohao Yan, Chengyu Shui.

   Background: Postoperative recurrence risk in cervical cancer remains heterogeneous, and conventional clinicopathological factors may not fully capture the contribution of the immune microenvironment and systemic inflammation. We investigated whether an integrated immune-inflammatory phenotype combining stromal tumor-infiltrating lymphocytes (TILs) and the systemic immune-inflammation index (SII) could improve recurrence-free survival (RFS) stratification after surgery.
Methods: This retrospective cohort study included 612 patients with cervical cancer who underwent primary surgery between January 2020 and December 2025. Stromal TILs were assessed on hematoxylin-eosin sections, and pretreatment SII was calculated from blood counts. An integrated immune-inflammatory phenotype was defined as favorable, poor, or intermediate. Kaplan-Meier analysis, restricted cubic spline modeling, and multivariable Cox regression were performed. Prognostic performance was compared across a clinical Cox model, an immune-extended Cox model, LASSO-Cox, CoxBoost, and random survival forest (RSF) using C-index, time-dependent area under the curve (AUC), integrated Brier score (IBS), calibration, and 36-month landmark decision curve analysis. Tissue-level validation was performed using immunohistochemical assessment of CD8, CD163, and PD-L1.
Results: During follow-up, 119 patients experienced an RFS event. Kaplan-Meier analysis showed significant RFS differences according to TIL category, SII category, and integrated phenotype. Restricted cubic spline analysis demonstrated a significant overall association between SII and recurrence risk, without marked nonlinearity. In multivariable analysis, FIGO IIIC disease, positive margin status, and the poor integrated phenotype remained independently associated with worse RFS. In model comparison, the immune-extended Cox model had the lowest IBS (0.278), LASSO-Cox achieved the highest C-index (0.782) and the best discrimination at 24 and 36 months, and RSF showed the highest 60-month AUC. In 36-month decision curve analysis, LASSO-Cox and RSF showed the greatest net benefit at intermediate thresholds. Immunohistochemical validation showed that the favorable phenotype was characterized by higher CD8+ cell density, lower CD163+ cell density, a higher CD8/CD163 ratio, and higher PD-L1 combined positive score.
Conclusion: An integrated immune-inflammatory phenotype combining stromal TILs and SII was independently associated with postoperative recurrence risk in cervical cancer and corresponded to distinct tissue immune states. This phenotype may provide a practical framework for recurrence risk stratification, while LASSO-Cox and RSF offer complementary prognostic perspectives.

Keywords:  LASSO-COX; cervical cancer; immunohistochemistry; integrated immune-inflammatory phenotype; random survival forest; recurrence-free survival; systemic immune-inflammation index; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3389/fimmu.2026.1850891
bioRxiv. 2026 Jun 24. pii: 2026.06.19.733403. [Epub ahead of print]

Circulating and brain-resident memory CD8+ T cells seed distinct bystander T RM -like populations in glioblastoma.

Sierra A Kleist, Tiffany Chen, Shawn C Musial, Neva R DiBlasi, Hanna N Degefu, Sarah C Berman, Myles A Ford, Jordan F Isaacs, Amanda Cruz Rivera, Arielle J Sclar, Christina V Angeles, Chun-Chieh Lin, Nathan E Simmons, Linton T Evans, Sladjana Skopelja-Gardner, Mary Jo Turk, Alexander G J Skorput, Sonia M Leach, Pamela C Rosato.

Across cancers, tumor-infiltrating CD8+ T cells expressing the tissue-resident memory T cell (T RM ) markers CD69 and CD103 are strongly associated with favorable clinical outcomes. However, a substantial fraction of these cells in human tumors are not tumor-specific, but instead recognize unrelated viral antigens. These virus-specific bystander T RM -like cells are prevalent in tumors and retain functional potential, raising interest in strategies that leverage pre-existing antiviral immunity for cancer immunotherapy. Yet their origins and differentiation states remain poorly defined, limiting both the interpretation of residency-based tumor-infiltrating lymphocyte (TIL) phenotyping and efforts to rationally harness these T RM -like cells. Here, using mouse models of GBM and melanoma, we demonstrate that resting circulating memory T cells trafficked into tumors via GPCR-dependent signaling and rapidly adopted a tissue-resident phenotype, independent of cognate antigen. Strikingly, in GBM, but not melanoma, pre-existing brain T RM contributed substantially to the bystander TIL compartment and were the dominant source of CD69+/CD103+ bystander T cells, revealing a tumor- and tissue-specific origin for this subset. These findings were further supported by transcriptional analysis of T cell receptor clones present in both paired patient GBM tumor and peritumoral brain, which identified shared features with T RM -derived TILs in mouse GBM. Overall, this work provides new insight into tumor immunosurveillance, inform the interpretation of CD69+/CD103- and CD103+ TIL populations, and lay a foundation for immunotherapeutic strategies aimed at harnessing circulating and pre-existing virus-specific T RM populations in tumors.

DOI: https://doi.org/10.64898/2026.06.19.733403
Front Immunol. 2026 ;17 1836156

Machine learning-driven identification and immunohistochemical validation of an integrated immune-inflammatory phenotype for disease-free survival stratification in breast cancer.

Shanshan Han, Lin Ran, Zhaoan Lian, Yong Tian, Li Qin, Yingchun Xiang, Xiaohao Yan, Chengyu Shui, Cheng Huang.

   Background: Recurrence risk in breast cancer remains heterogeneous, and conventional clinicopathological variables may not fully capture the contribution of immune-related factors. We compared multiple survival modeling strategies and evaluated whether an integrated immune-inflammatory phenotype could improve disease-free survival (DFS) stratification.
Methods: This retrospective single-center study included 503 patients with surgically treated breast cancer between January 2020 and December 2025. Stromal tumor-infiltrating lymphocytes (TILs) were assessed from pathological sections, and the systemic immune-inflammation index (SII) was calculated from pre-treatment blood counts. An integrated immune phenotype was defined as favorable (high TILs/low SII), poor (low TILs/high SII), or intermediate (all remaining combinations). A base clinical Cox model, an immune-extended Cox model, LASSO-Cox, CoxBoost, and random survival forest (RSF) were compared using C-index, time-dependent area under the curve (AUC), integrated Brier score (IBS), and decision curve analysis. Conventional survival analyses, restricted cubic spline analysis, and immunohistochemical validation with CD8 and CD163 staining were also performed.
Results: During follow-up, 107 patients (21.3%) experienced a DFS event. RSF achieved the best overall performance, with time-dependent AUCs of 0.867, 0.880, 0.879, 0.893, and 0.911 at 12, 24, 36, 48, and 60 months, respectively, and the lowest IBS (0.100). In the RSF model, pathological N stage was the most important predictor, followed by SII, integrated immune phenotype, Ki-67, and lymphovascular invasion. Kaplan-Meier analysis showed no significant DFS difference according to TIL category alone, whereas high SII and the poor integrated immune phenotype were associated with significantly worse DFS. In the final multivariable Cox model, the poor phenotype remained independently associated with worse DFS compared with the favorable phenotype (hazard ratio 2.53, 95% confidence interval 1.39-4.60; p = 0.002). Immunohistochemical validation showed higher CD8+ cell density, lower CD163+ cell density, and a higher CD8/CD163 ratio in the favorable phenotype than in the poor phenotype.
Conclusion: RSF provided the best prognostic performance in this cohort. SII and the integrated immune phenotype emerged as clinically relevant predictors, and the integrated phenotype showed tissue-level biological support. Combining machine learning-based survival modeling with pragmatic immune-inflammatory markers may improve recurrence risk stratification in breast cancer.

Keywords:  breast cancer; disease-free survival; integrated immune phenotype; random survival forest; systemic immune-inflammation index; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3389/fimmu.2026.1836156
Lung India. 2026 Jul 01. 43(4): 404-411

A pragmatic workflow for advanced NSCLC diagnosis in routine practice: Integrating histopathology, PD-L1 scoring, and EGFR testing.

Ashmeet Kaur, Ranjana Solanki, Ravindra Singh Gothwal, Arpita Jindal.

   BACKGROUND: Precision treatment of advanced non-small cell lung carcinoma (NSCLC) requires early biomarker availability, yet routine practice is constrained by small biopsies, sequential testing, and limited resources. We assessed whether PD-L1 and EGFR testing were completed from the initial biopsy using a tissue-efficient reflex workflow integrating histologic subtyping, PD-L1 scoring, EGFR analysis, and tumour-infiltrating lymphocyte (TIL) assessment.
METHODS: In this prospective single-arm feasibility study, 50 stage III or IV NSCLC cases were evaluated. Tumours were subtyped using morphology and minimal immunohistochemistry (TTF-1, Napsin A, p40). PD-L1 immunohistochemistry was performed using a locally validated CAL10-based laboratory-developed test and scored by tumour proportion score (TPS) and combined positive score (CPS). Reflex EGFR testing was attempted in adenocarcinomas with adequate tissue. TILs were graded morphologically. Tissue adequacy and biomarker associations were analysed.
RESULTS: Squamous cell carcinoma comprised 56% and adenocarcinoma 44% of cases. Adequate tissue to complete PD-L1 and EGFR testing was available in 59% (13/22) of adenocarcinomas, and was reported within 30 days. EGFR mutations were detected in 31% of tested adenocarcinomas and were associated with TPS <50% and absent TILs. High PD-L1 expression (TPS ≥50%) was observed in 14% of tumours, exclusively in EGFR wild-type cases. TILs were present in 12% of cases and were significantly associated with higher CPS ( P = 0.002), exceeding TPS.
CONCLUSION: A biopsy-level reflex workflow enables feasible, tissue-efficient concurrent molecular and immune profiling in advanced NSCLC. Integration of TPS, CPS, TIL assessment, and EGFR testing within a reflex workflow demonstrates the feasibility of coordinated immuno-molecular profiling from limited biopsy tissue in advanced NSCLC.

Keywords:  Advanced non-small cell lung carcinoma; EGFR mutation; PD-L1; combined positive score; laboratory-developed test; reflex testing; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.4103/lungindia.lungindia_835_25
Front Immunol. 2026 ;17 1876390

Dominant T cell receptor clonotypes in adrenocorticotropic hormone-secreting pituitary carcinoma are the highest-frequency clones among CD4+ and CD8+ cells in peripheral blood during effective anti-PD-1 therapy.

Mariko Sugiyama, Shintaro Iwama, Kazuhito Takeuchi, Tomoko Kobayashi, Takanori Murase, Daisuke Hagiwara, Hao Chen, Yuichi Nagata, Ryuta Saito, Yoshiki Akatsuka, Hiroshi Arima.

   Context: Pituitary carcinoma is a rare and highly aggressive tumor. While anti-programmed cell death-1 (PD-1) therapy has shown efficacy in some cases, the factors that predict a favorable response remain largely unclear.
Objective: To evaluate tumor-infiltrating lymphocytes (TILs) in pituitary carcinoma and to compare T-cell receptor (TCR) clonotypes between the pituitary and peripheral blood.
Methods: A 34-year-old woman with Lynch syndrome and adrenocorticotropic hormone-secreting pituitary carcinoma with hepatic metastasis received anti-PD-1 therapy, achieving durable disease control exceeding 1 year. Immunohistochemistry was performed on treatment-naïve surgical tumor samples, and TCR repertoire analyses were conducted on both the tumor sample and peripheral blood mononuclear cells collected during effective anti-PD-1 therapy.
Results: Treatment-naïve pituitary carcinoma tissues exhibited infiltration of CD4+ and CD8+ T cells. Analysis of the TCR repertoire identified 15 clonotypes with a high frequency (> 1% of sequencing reads) in the tumor; among these, four of the five most prevalent clonotypes were co-detected as dominant clones in peripheral blood after treatment, including the most abundant clones found within the CD4+ and CD8+ T cell populations. Despite control of the primary and hepatic lesions, ovarian metastasis developed, which was associated with reduced CD4+ TILs.
Conclusions: The presence of CD4+ and CD8+ TILs may underlie the immunological foundation for PD-1 blockade efficacy in pituitary carcinoma, supported by the detection of tumor-resident TCR clonotypes in peripheral blood during a positive therapeutic response.

Keywords:  Cushing’s syndrome; PD-1; T−cell receptor repertoire; immunotherapy; pituitary cancer

DOI:  https://doi.org/10.3389/fimmu.2026.1876390
Discov Oncol. 2026 Jul 02.

Engineered immune cells and extracellular vesicles target tumour microenvironment barriers in solid tumour immunotherapy.

Xiaowei Zhou, Shixin Chen, Junbo Liang.

  Adoptive cell therapy (ACT) for solid tumours frequently fails because the tumour microenvironment (TME) imposes multiple, overlapping barriers, including stromal exclusion, suppressive myeloid networks, inhibitory cytokines and metabolites, and antigen heterogeneity. Collectively, these factors markedly restrict the infiltration, persistence, and cytotoxic function of effector lymphocytes. In this Review, we synthesise recent primary studies, consensus guidance, and clinical-trial evidence on engineered-cell therapies, including chimeric antigen receptor (CAR) T cells, T-cell receptor-engineered T cells (TCR-T cells), CAR-NK cells, CAR-macrophages (CAR-M), and emerging in vivo CAR-engineering strategies, together with extracellular-vesicle (EV)-based therapeutics. We then map each platform to mechanism-linked resistance nodes within the TME. For engineered cells, key design levers include context-restricted recognition to reduce on-target/off-tumour toxicity, resistance to dominant suppressive pathways such as TGF-β and adenosine signalling, improved trafficking and tissue penetration, and controllability through transient programming or pharmacological switches. For EVs, the main translational advantages include tissue penetration, modular surface engineering, cargo loading, and their acellular nature, which avoids risks related to in vivo cellular expansion but introduces distinct challenges such as rapid clearance, immunogenicity, batch heterogeneity, and uncertain potency assays. Early clinical data using KRAS G12D-targeting engineered exosomes in metastatic pancreatic cancer support the feasibility of this approach and suggest that EVs may also remodel the immune microenvironment, providing a rationale for combination strategies. We propose a barrier-matched framework in which engineered cells and extracellular vesicles are assigned as functionally orthogonal but complementary modules: engineered cells provide adaptive cytotoxicity, whereas EVs enable microenvironmental reconditioning. This framework may help guide rational combination strategies designed to systematically dismantle resistance in solid tumours.

Keywords:  CAR-NK; CAR-T; CAR-macrophage; Engineered exosomes; Extracellular vesicles; Immunotherapy resistance; Solid tumours; Tumour microenvironment

DOI:  https://doi.org/10.1007/s12672-026-05523-x
Ther Adv Med Oncol. 2026 ;18 17588359261456794

Homologous recombination deficiency correlates with tumor-infiltrating lymphocytes and predicts patient outcomes in hormone receptor-positive/HER2-negative breast cancer.

Shu-Jie Zhang, Jian-Fei Fu, Cui-Cui Li, Lan Wang, Xin Zhao, Xin-Yu Zhu, Zi-Liang Qian, Wen-Yong Sun, Lin-Feng Zheng, Xiang-Yi Chen, Xiang Lu, Wen-Ming Cao.

   Background: Hormone receptor-positive/human epidermal growth factor receptor 2-negative (HR+/HER2-) breast cancer represents the most prevalent molecular subtype but demonstrates pronounced biological and clinical heterogeneity. Homologous recombination deficiency (HRD) and tumor-infiltrating lymphocytes (TILs) have recently emerged as key determinants of prognosis and immune activity, but their interplay in this subtype remains understudied.
Objectives: To assess the relationship between HRD, TILs, and clinical outcomes in HR+/HER2- breast cancer and validate the prognostic value of HRD.
Design: Retrospective multicenter cohort study.
Methods: A total of 365 patients (332 with available TILs data) from three institutions were enrolled. HRD was quantified using Shallow HRD algorithm on low-depth whole-genome sequencing (threshold: score ⩾6). TILs were evaluated by the MD Anderson system (⩾10% classified as high). Spearman correlation, Kaplan-Meier survival analysis, and multivariable Cox regression were performed.
Results: HRD scores were inversely correlated with TILs (Spearman rho = -0.13, p = 0.031). HRD-high patients had significantly inferior 10-year survival (invasive disease-free survival (IDFS): 31.0% vs 57.9%, hazard ratio (HR) = 2.36; distant recurrence-free survival (DRFS): 34.1% vs 61.1%, HR = 2.48; overall survival (OS): 48.7% vs 79.0%, HR = 2.79; p < 0.0001). HRD was confirmed as an independent prognostic predictor (IDFS HR = 2.03, DRFS HR = 2.16, OS HR = 2.15; p < 0.01). No significant survival benefit from anthracycline-based chemotherapy was observed in HRD-high tumors (HR = 1.32, p = 0.451). High TILs showed no significant association with survival outcomes in this cohort.
Conclusion: HRD represents a reliable independent prognostic biomarker in HR+/HER2- breast cancer in this cohort, with an inverse association with TILs suggesting immune evasion. Its potential impact on prognosis and treatment requires further validation in prospective clinical trials.

Keywords:  HR+/HER2− breast cancer; homologous recombination deficiency; prognosis; shallow HRD algorithm; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.1177/17588359261456794
Front Immunol. 2026 ;17 1819470

Fusion protein-mediated costimulation in engineered T cells: from intrinsic signaling to tumor microenvironment rewiring.

Ryma Toumi, Simonne J Guenette, Shannon K Oda.

  Adoptive T cell therapies have markedly improved outcomes in hematologic malignancies but their efficacy in solid tumors can be diminished by a hostile tumor microenvironment that impedes sustained therapeutic responses. Beyond challenges such as limited trafficking and antigen heterogeneity, engineered T cells face suppressive myeloid and stromal populations, inhibitory checkpoint ligand interactions, and metabolically hostile niches that collectively diminish effector function and persistence. To overcome these barriers, a new generation of fusion protein-based costimulatory strategies has emerged that couple ligand-guided sensing of the tumor microenvironment with modular control of T cell activation and fate. This review examines how conventional and non-canonical costimulatory modules, when incorporated into chimeric antigen receptor (CAR) and T cell receptor (TCR) architectures, modulate T cell differentiation and function within the tumor site. It further analyzes how membrane-anchored and secreted fusion proteins enable engineered T cells to activate dendritic cells, reprogram myeloid cells, and convert poorly inflamed tumors into treatment-responsive environments. Together, these advances establish a design framework in which fusion protein-based receptors and ligands enhance T cell function and remodel the tumor microenvironment, thereby expanding the therapeutic potential of adoptive T cell therapy for solid tumors.

Keywords:  T cell engineering; costimulatory signaling; fusion protein; myeloid cell reprogramming; switch receptor; tumor microenvironment

DOI:  https://doi.org/10.3389/fimmu.2026.1819470
J Immunother Cancer. 2026 Jul 01. pii: e014518. [Epub ahead of print]14(7):

PARP inhibition combined with a T-cell receptor β chain-directed antibody fusion molecule drives polyclonal antitumor immunity and tumor regression.

Ginette Santiago-Sanchez, Francesca Rosato, Kellsye P Fabian, Michelle R Padget, Jung-Min Lee, Fatima Karzai, Jeffrey Schlom, James L Gulley, Duane H Hamilton, Jonelle K Lee, Margaret R Pruitt, Andrew Bayliffe, Zhen Su, Jacques Moisan, Madan Katragadda, James W Hodge.

   BACKGROUND: Metastatic castration-resistant prostate cancer (mCRPC) remains an aggressive disease with limited response to systemic therapies despite androgen deprivation. Immune-excluded prostate tumors are typically resistant to immunotherapy. STAR0602 is a selective bifunctional T-cell agonist composed of an antibody targeting Vβ6 and Vβ10 T-cell receptor chains fused to human interleukin-2 that selectively expands Vβ6+ CD8+ memory T cells and has demonstrated clinical activity as a monotherapy in anti-programmed death-ligand 1-resistant tumors (NCT05592626). We hypothesized that combining poly ADP-ribose polymerase (PARP) inhibition with Vβ-directed T-cell activation would enhance antitumor immunity and promote polyclonal T-cell responses in immune-excluded prostate cancer.
METHODS: The antitumor activity of the PARP inhibitor olaparib combined with mSTAR1302, the murine surrogate of STAR0602, was evaluated in TRAMP-C2 and RM-1 prostate tumor models. Tumor growth, survival, and immune responses were assessed by flow cytometry and functional depletion studies. The role of tumor-intrinsic TRAIL-R2 signaling was evaluated using a TRAIL-R2 knockout TRAMP-C2 model, and clinical relevance was explored by assessing TRAIL-R2 expression in tumor samples from patients treated with olaparib in a Phase 2 clinical trial (NCT02484404).
RESULTS: Combination therapy with olaparib and mSTAR1302 induced significant tumor regression and improved survival compared with either agent alone. Treatment increased tumor-infiltrating lymphocytes, expanded activated Vβ13+ CD4+ and Vβ13+ CD8+ T cells, reduced immunosuppressive populations, and enriched stem-like progenitor exhausted CD8+ T cells. Depletion studies demonstrated that Vβ13+ CD4+ T cells, Vβ13+ CD8+ T cells, natural killer cells, and interferon-γ were required for therapeutic efficacy. TRAIL-R2 knockout experiments demonstrated that tumor-intrinsic TRAIL-R2 signaling contributes to the antitumor activity of the combination. Mechanistically, these findings support a model in which PARP inhibition sensitizes tumors while mSTAR1302-mediated Vβ13+ T-cell expansion initiates a broader polyclonal antitumor immune response associated with antigen spreading and recognition of multiple tumor antigens and neoepitopes.
CONCLUSIONS: These findings suggest that combining tumor-sensitizing therapies with selective T-cell activation may represent a broader strategy to overcome immune exclusion in solid tumors. Together, these results provide mechanistic rationale for clinical evaluation of olaparib in combination with STAR0602 in patients with mCRPC who have progressed on androgen deprivation therapy.

Keywords:  Combination therapy; Immune modulatory; Immunotherapy; Prostate Cancer; T cell

DOI:  https://doi.org/10.1136/jitc-2025-014518
Acta Pharm Sin B. 2026 Jun;16(6): 3892-3906

Interleukin 12-tethered nano-aluminum adjuvant orchestrates environmental immunomodulation to empower cytotoxic T cells against solid tumors.

Aining Shen, Jing Zhao, Zhenwei Su, Ran Wang, Ying Li, Chaojie Zhu, Qingyuan Zhao, Pinwan Yu, Zhi Ping Xu, Lingxiao Zhang.

  The immunosuppressive tumor microenvironment (TME) profoundly limits the therapeutic efficacy of CD8+ T cells in solid tumors. While cytokine therapies have shown promise in reactivating CD8+ T cells, they fail to address the common suppressive environmental attributes (e.g., acidosis and Mg2+ deficiency) of solid tumors. Here, we report an innovative CD8+ T cell dual-functional modulator, interleukin-12-tethered nano-aluminum adjuvant (IL12@NAM), which counteracts the acidic TME to relieve acidosis and concurrently releases Mg2+ and IL12. Locally released Mg2+ and IL12 synergize in T cell infiltration and activation by promoting the phosphorylation of focal adhesion kinase and extracellular signal-regulated kinase 1/2, and enhance CD8+ T cell activation and functions via the Ca2+-nuclear factor of activated T cells 2 pathway. Furthermore, dual-functional IL12@NAM mitigates CD8+ T cell exhaustion by reducing PD1 and LAG3 expression and effectively increases the differentiation towards T helper 1 cells while decreasing regulatory T cells, creating a more favorable immune network for enhanced CD8+ T cell-mediated anti-tumor immunity. As a result, IL12@NAM has demonstrated potent therapeutic efficacy against advanced melanoma and breast cancer, and remarkably empowered adoptive T therapy of solid tumors. This study provides a paradigm for empowering cytotoxic T cells by reactivating and creating a sustainable immunoresponsive environment, offering a potential adjuvant strategy to enhance solid tumor therapy.

Keywords:  Acidosis alleviation; Cytotoxic T cell activation; Cytotoxic T cell immune exhaustion; Cytotoxic T cell infiltration; Interleukin-12 tumor therapy; Mg2+ supplementation; Nano-aluminum adjuvants; Tumor microenvironment

DOI:  https://doi.org/10.1016/j.apsb.2025.10.021
Sci Adv. 2026 Jul 03. 12(27): eaea4727

Single-cell transcriptomic analysis reveals tumor-immune determinants of lymph node colonization and progression in thyroid cancer.

Anthony T Nguyen, Jolene Viramontes, Isaiah Vazquez, Catriona McWilliam, Vaishnavi Devarakonda, Regina Henson, Wendy L Sacks, Jon Mallen-St Clair, Yufei Chen, Evan Walgama, Kevin S Scher, Justin Moyers, Howard M Sandler, Julie K Jang, Zachary S Zumsteg, Wonwoo Shon, Stephen L Shiao, Allen S Ho.

Lymph node (LN) metastases are a major driver of mortality across solid cancers, including thyroid carcinomas, which are known for high rates of nodal colonization. To elucidate the determinants of nodal spread, we isolated tumor-infiltrating leukocytes from primary thyroid tumors and matched metastatic LNs for single-cell RNA sequencing with validation by multiplex immunohistochemistry. Comparing the microenvironmental alterations between primary tumors and their LNs, we found that thyrocytes and tumor-associated macrophages down-regulate the expression of multiple inflammatory cytokine receptors, including TNFRSF12A and CX3CR1, upon LN colonization. LNs were associated with the induction of regulatory T cells to suppress T cell-mediated cytotoxicity compared to matched primary tumors. Notably, tumor-infiltrating lymphocytes within LNs demonstrated increased expression of activation markers, including interleukin-7 receptor (IL7R). High LN expression of IL7R was significantly correlated with improved outcomes and can serve as a biomarker in this heterogeneous disease. Our findings on the dynamic equilibrium within LN metastases may offer conserved mechanisms for nodal colonization across solid tumors.

DOI: https://doi.org/10.1126/sciadv.aea4727
Signal Transduct Target Ther. 2026 Jul 01. pii: 255. [Epub ahead of print]11(1):

Advances and prospects in cell therapy for cancer: explorations from T cells to stem cells.

Guisha Zi, Lei Zhang, Ling Zhou, Xiansheng Liu, Lingling Wang, Runxuan Zhou, Pengdou Zheng, Jia Wei, Xiaoping Chen, Shuang Wei.

The increasing global burden of cancer necessitates innovative therapeutic strategies. Cell therapy represents a major breakthrough in oncology, evolving rapidly from the successful application of chimeric antigen receptor T (CAR-T) cells in hematologic malignancies to a multiplatform landscape characterized by the concurrent development of diverse strategies. Current research focuses on T cell receptor-engineered T (TCR-T) cells, tumor-infiltrating lymphocytes (TILs), gamma delta (γδ) T cells, CAR-natural killer (CAR-NK) cells, CAR-macrophages (CAR-Ms), and various strategies based on dendritic cells (DCs), B cells, and stem cells. The translational paradigm is expanding from the relatively mature field of hematologic malignancies to the more prevalent and mechanistically complex domain of solid tumors. In recent years, this field has exhibited a clear trend toward expansion from autologous therapies to allogeneic "off-the-shelf" platforms. Approaches such as CAR-NK and CAR-natural killer T (CAR-NKT) cell therapies exhibit significant clinical potential because of their low immunogenicity and reduced risk of graft-versus-host disease (GvHD). Concurrently, in vivo engineering technologies that directly deliver CAR genes in situ are emerging as promising approaches to lower costs and simplify manufacturing by bypassing complex ex vivo procedures. This review systematically outlines recent advances in these strategies, focusing on their mechanisms of action, target antigens, and clinical translation. Despite progress, formidable challenges remain, including tumor heterogeneity, the immunosuppressive tumor microenvironment (TME), and therapy-related toxicity. To address these challenges, future research will focus on novel target discovery, enhanced toxicity management, and scalable manufacturing processes. The integration of multidisciplinary technologies, such as multiomics analysis, artificial intelligence, and synthetic biology, will advance cell therapies toward safer, more effective, and widely accessible applications.

DOI: https://doi.org/10.1038/s41392-026-02780-8
Clin Cancer Res. 2026 Jun 30.

A randomized phase 2 study of combination atezolizumab and varlilumab (CDX-1127) with or without cobimetinib in previously-treated unresectable biliary tract cancer.

Thatcher Ross Heumann, Jiayun Lu, Hao Wang, Mitch Phelps, Qingfeng Zhu, Robert Anders, Sarah Mitchell, James Leatherman, Nicole Abbott, Kyeongmin Kim, Teresa Imhof, Zhiliang Xie, Alexandra Partey, Daneng Li, Paul Raymond Kunk, Renuka V Iyer, Farshid Dayyani, Lionel Aurelien Kankeu, Aparna Kalyan, Olumide B Gbolahan, Milind M Javle, S Lindsey Davis, Vaia Florou, Kristen Spencer, Elad Sharon, Mark Yarchoan, Gregory B Lesinski, Nilofer Saba Azad.

PURPOSE: The addition of MEK inhibition to PD-L1 blockade improves progression-free survival (PFS) in patients with advanced biliary tract cancer (BTC). While MEK inhibitors may increase tumor cell immunogenicity, they can impair T-cell priming/effector function, limiting combination efficacy. We hypothesized that the addition of a CD27 agonist could restore T-cell function and enhance anti-tumor immunity in this combination.
PATIENTS AND METHODS: We conducted a randomized, phase 2 trial evaluating atezolizumab (840mg IV days 1,15) in combination with the CD27 co-stimulatory mAb (CDX-1127/varlilumab [3mg/kg IV days 1, 15]), with/without the addition of a MEK inhibitor (cobimetinib [60mg oral daily days 1-21, off 22-28]) in unresectable BTC following at least 1 metastatic therapy. Overall response rate (ORR) and PFS were co-primary endpoints. Treatment-related changs in CD8+ tumor infiltrating lymphocytes (TIL) was the primary correlative outcome.
RESULTS: The trial was closed early following interim preplanned ORR analysis. At closure, 57 patients had been enrolled (n=29[CAV],n=28[AV]. A majority (67%) had intrahepatic cholangiocarcinoma and 32% were immunotherapy-experienced. Both regimens were well-tolerated without new safety signals. Objective responses were rare (0%[CAV],3.8%[AV]). Median PFS was 2.40(CAV) and 1.84(AV) months (HR 0.67,95%CI[0.38,1.18]). Among immunotherapy-experienced patients, mPFS was 3.62(CAV) and 1.84(AV) months (HR 0.54,95%CI:[0.18-1.62]).  Treatment with CAV increased intratumoral CD8+ T-cell density compared with treatment with AV.
CONCLUSIONS: The combinations of atezolizumab and varlilumab with/without cobimetinib were safe but neither meaningfully improved outcomes in BTC treated in the later lines. Correlative tissue studies validated preclinical work that MEK inhibition increases CD8+TILs.

DOI: https://doi.org/10.1158/1078-0432.CCR-25-4893
Proc Natl Acad Sci U S A. 2026 Jul 07. 123(27): e2602385123

Sustained A2AR expression and loss paradoxically promote CD8+ T cell exhaustion.

Longzhen Song, Arjun Kharel, Ping Xie, Jie Fan, Abu Baker, Yuqi Zhang, Yi Zhang, Weiguo Cui, Bin Zhang.

  Although A2AR is a key immunoregulatory receptor that suppresses CD8+ T cell activation in response to elevated extracellular adenosine in inflamed or hypoxic microenvironments, its role in CD8+ T cell differentiation and cell-fate decisions during chronic viral infection and cancer remains poorly understood. Using A2AR-eGFP reporter mice, we show that A2AR expression is rapidly induced by TCR stimulation and persists under chronic antigen exposure and hypoxia, with sustained expression strongly associated with terminal exhaustion via the canonical Gαs-cAMP-PKA pathway. Paradoxically, A2AR loss does not alleviate exhaustion but instead accelerates differentiation toward the terminally exhausted state. Single-cell multiomics profiling revealed that A2AR deficiency activates CD122 (IL-2Rβ)-dependent signaling, driving T cell exhaustion. Genetic deletion of CD122 in A2AR-deficient CD8+ T cells reduced terminal exhaustion, identifying CD122 signaling as a key mediator of A2AR loss-driven exhaustion. Intriguingly, both sustained A2AR expression and A2AR loss converge to promote T cell exhaustion differentiation through distinct mechanisms. These findings uncover a paradoxical role of A2AR in shaping CD8+ T cell fate choices during chronic infection and cancer.

Keywords:  A2AR; CD8(+) T cell differentiation; exhaustion; immunotherapy

DOI:  https://doi.org/10.1073/pnas.2602385123
Funct Integr Genomics. 2026 Jun 29. pii: 169. [Epub ahead of print]26(1):

Single-cell transcriptomic analysis identifies a tumor-enriched GIMAP7⁺ CD4⁺ naïve T cell population with a unique immunoregulatory state in lung adenocarcinoma.

Zhirong Yang, Shangzhe Zheng, Shiyu Liu, Yiming Zeng, Jie Li, Xi Chen, Yuan Xu.

  Lung adenocarcinoma (LUAD) is characterized by complex immune microenvironments that modulate disease progression and response to therapy. CD4⁺ naïve T cells are traditionally considered precursors of effector T cells, yet their functional states within LUAD remain poorly defined. We integrated single-cell RNA sequencing (scRNA-seq) data from LUAD tumor and distal normal tissues with bulk RNA-seq datasets (GSE159857 and TCGA-LUAD) to characterize CD4⁺ naïve T cell heterogeneity. Subclustering, trajectory analysis, cell-cell communication inference, and transcription factor network analysis were applied to identify tumor-associated states. qPCR was conducted to validate the differential expression of key transcription factors and hub genes in LUAD tissues. CD4⁺ naïve T cells were selectively expanded in LUAD and transcriptionally reprogrammed. A GIMAP7-high subpopulation was enriched in tumors, occupied early pseudotime states, and exhibited elevated LTB expression with distinct CCR6 dynamics. Functional enrichment analyses revealed upregulation of cytokine-mediated signaling and T cell receptor pathways, alongside downregulation of cytotoxic programs. Cell-cell communication analysis demonstrated enhanced interactions of GIMAP7⁺ cells via MHC-I, MHC-II, and Cyclophilin A (CypA) pathways. SCENIC analysis revealed downregulation of key transcription factors (EOMES, TBX21, SPI1), suggesting constrained effector differentiation. Bulk RNA-seq validation confirmed LTB upregulation and expansion of GIMAP7⁺ CD4⁺ naïve T cells. qPCR confirmed the decreased expression of TBX21 and SPI1, along with elevated levels of LTB and LTA in LUAD tissues. Our study identifies a tumor-enriched GIMAP7⁺ CD4⁺ naïve T cell population in LUAD that defines an immunoregulatory T cell state characterized by LTB-driven signaling, enhanced intercellular communication, and impaired effector differentiation. These findings provide new insights into tumor-immune interactions and highlight potential targets for immunomodulatory therapies.

Keywords:  CD4⁺ naïve T cells; GIMAP7; LTB; Lung adenocarcinoma; Single-cell RNA sequencing; Tumor microenvironment

DOI:  https://doi.org/10.1007/s10142-026-01945-6
Oncol Lett. 2026 Aug;32(2): 356

CD47 monoclonal antibody enhances the inhibitory effect of anti-HER2 chimeric antigen receptor macrophages on ovarian cancer.

Jie Qiao, Qing Kou, Ji Qiu.

  Macrophages have garnered notable interest as therapeutic vehicles due to their innate phagocytic ability, tumor tropism, and pivotal role in linking innate and adaptive immunity. These attributes have led to the development of various macrophage-centered treatments. Among them, chimeric antigen receptor macrophages (CAR-Ms) have emerged as a promising adoptive cell therapy, demonstrating potential for clinical application in multiple solid tumors. However, their effector functions remain amenable to further enhancement. In the present study, anti-HER2 CAR-Ms were constructed using an adenoviral vector system. The specificity and antitumor efficacy against HER2+ ovarian cancer cells were assessed using flow cytometry. The effects of a CD47 monoclonal antibody (mAb) on CAR-M-mediated killing and phenotypic polarization were subsequently investigated. Additionally, the combined therapeutic benefit of CD47 mAb and anti-HER2 CAR-Ms was evaluated in a murine subcutaneous xenograft model of ovarian cancer. Anti-HER2 CAR-Ms specifically recognized and killed HER2-expressing ovarian cancer cells. The addition of CD47 mAb enhanced the phagocytic capacity, promoted a pro-inflammatory phenotype and inflammatory cytokine secretion in CAR-Ms. In vivo, combination therapy significantly reduced tumor burden in xenograft-bearing mice. Collectively, these findings indicate that the combination of CAR-Ms with CD47 blockade offers a robust and promising strategy for personalized targeted therapy in solid tumors.

Keywords:  CD47 monoclonal antibody; chimeric antigen receptor; immunotherapy; macrophage

DOI:  https://doi.org/10.3892/ol.2026.15711
Cancer Immunol Immunother. 2026 Jun 30.

R-5780, a SagA-engineered Lactococcus lactis, is a safe oral synthetic-biology microbial therapy that potentiates PD-1 blockade.

Miaki Fukuhara, Bikash Sahay, Gary R Fanger, Christian Furlan Freguia.

  Immune checkpoint inhibitors (ICIs) targeting programmed cell death protein 1 (PD-1) have transformed cancer therapy, yet response rates remain limited across solid tumors. Modulation of the gut microbiome has emerged as a promising strategy to enhance immunotherapy efficacy. R-5780 is an engineered Lactococcus lactis strain expressing the peptidoglycan hydrolase Secreted Antigen A (SagA), which generates muramyl dipeptide (MDP), a natural ligand of the nucleotide-binding oligomerization domain 2 (NOD2) receptor involved in innate immune activation and antigen-presenting cell maturation. Therapeutic potential of R-5780 was assessed in vivo murine models. Using the CT26 murine colorectal carcinoma model, oral administration of R-5780 with anti-PD-1 significantly inhibited tumor growth in both prophylactic and therapeutic settings compared to ICI therapy alone. Mechanistic studies revealed that R-5780 enhanced dendritic-cell activation and promoted a pro-inflammatory tumor microenvironment characterized by elevated interleukin-1 beta (IL-1β). Multiplex cytokine profiling and flow cytometry further showed that R-5780 reduced CD8+ T-cell exhaustion, with decreased expression of PD-1, LAG-3, and TIM-3 on intratumoral T cells. In murine toxicology studies, R-5780 was well tolerated, with no adverse events or systemic inflammation observed following oral administration. Together, these results identify R-5780 as a safe and potent NOD2 activator that synergizes with PD-1 blockade to enhance antitumor immunity and support its advancement toward clinical evaluation.

Keywords:  Gut microbiome; Live biotherapeutic products; NOD2 signaling; PD-1 blockade; Preclinical toxicology; T-cell exhaustion

DOI:  https://doi.org/10.1007/s00262-026-04480-2
Front Immunol. 2026 ;17 1900363

Editorial: Exploring unconventional T cells and CAR-T-cells in cancer immunotherapy.

Laura Patrussi, Chenran Zhang.



Keywords:  CAR T cell; MAIT (mucosal-associated invariant T) cell; gamma delta T (γδ T) cells; iNKT (invariant natural killer T cell); tumor microenvironment - TME; unconventional T cell

DOI:  https://doi.org/10.3389/fimmu.2026.1900363
bioRxiv. 2026 Jun 17. pii: 2026.06.15.732401. [Epub ahead of print]

NKG2C + CD27 - Defines Human CD8 + Regulatory T Cells.

Xuyang Li, Somen K Mistri, Tian Cao, Rajesh K Krishnan, Martin Hemberg, Howard L Weiner, Dan Hu.

Here we identify NKG2C + CD27 - as a novel surface marker unifying multiple previously reported CD8 + regulatory T cell (Treg) populations. This population displays high clonality and divides into CD226 - (Treg1) and CD226 + (Treg2) subsets, with Treg2 exhibiting stronger suppressive activities. Up to 35% of CD8 + T EMRA cells are Tregs, whereas approximately 85% of CD8 + Tregs are T EMRA cells, which increase with aging. These findings establish a unified and novel cell surface marker for CD8 + Tregs and their subsets, which resolves prior heterogeneity in the field, and show that CD8 + T EMRA cells are a heterogeneous population that includes T cells with regulatory function. Our findings provide a critical framework for the isolation and in-depth functional characterization of CD8 + Tregs in health, aging, and disease.

DOI: https://doi.org/10.64898/2026.06.15.732401
Front Immunol. 2026 ;17 1762600

Programmed cell death ligand 1 in correlation with BRAFV600E mutation, molecular characteristics and biological behavior in ameloblastoma.

Aijing Pan, Zhipeng Xie, Jianhui Zhan, Yifeng Zhao, Yue Du, Kai Zhang.

   Objective: To investigate the expression of Programmed cell death ligand 1 (PD-L1) in Ameloblastoma (AM) and its correlation with BRAFV600E mutation as well as the expression of Cytotoxic CD8+ T cells (CD8+ T) and Forkhead box protein 3 (FoxP3) in the immune microenvironment. And to preliminarily explore the potential association of PD-L1 with the biological behavior and Disease-free survival (DFS) of patients with AM, so as to provide a theoretical reference for clinical diagnosis and treatment.
Methods: Fifty formalin-fixed paraffin-embedded (FFPE) specimens of AM were enrolled in this study. Immunohistochemistry (IHC) was performed to detect the expression of BRAFV600E mutant protein, CD8+ T cells, FoxP3+ T cells and PD-L1. Molecular testing for the BRAFV600E mutation was further conducted in 29 eligible AM specimens. PD-L1 expression was evaluated by IHC, and the combined positive score (CPS) ≥ 1 was defined as PD-L1 positivity. The chi-square test and Fisher's exact test were used for bivariate statistical analysis. Survival analysis was performed using the Kaplan-Meier method with the log-rank test, and the Cox proportional hazards regression model was applied to identify predictive factors for DFS.
Results: Of the 50 patients, 42 (84%) were PD-L1 positive. Forty-one cases were BRAF mutation-positive, and 38 of them (92.68%) showed PD-L1 positivity, indicating a significant association between the two (p = 0.003). No significant correlations were found between PD-L1 and other major indicators, including CD8+ T cells (p = 0.247), FoxP3+ cells (p = 0.702), and pathological subtype (p = 0.667). In exploratory survival analysis, patients with high CD8+ T cell infiltration exhibited a tendency of prolonged DFS compared with those with low infiltration (HR = 0.112, 95%CI: 0.014-0.913, p = 0.041).
Conclusion: PD-L1 expression is frequently observed in AM with BRAFV600E mutation, whereas no significant correlation is identified between PD-L1 and the clinicopathological parameters associated with AM. Within the limitation of relatively short follow-up duration, high CD8+T cell infiltration was indicated to be potentially correlated with favorable DFS in AM patients, which needs further verification in cohorts with longer follow-up.

Keywords:  CD8+ T lymphocytes; PD-L1; ameloblastoma; immunotherapy; molecular targeted therapy

DOI:  https://doi.org/10.3389/fimmu.2026.1762600
Inflammation. 2026 Jul 01.

Opposing Dynamics of CD4 + Regulatory and CD8 + Foxp3+ T Cells Characterize Immune Imbalance in Rheumatoid Arthritis.

Baochen Li, Juanjuan Liu, Jing Li, Ruihe Wu, Yuxin Fan, Chong Gao, Caihong Wang.

  Rheumatoid arthritis (RA) is a chronic autoimmune disease marked by persistent inflammation and joint damage from impaired immune tolerance. While CD4 + regulatory T cells (Tregs) are key to immune balance, their ability to restore tolerance in RA remains insufficient. Recently, CD8 + Foxp3+T have emerged as distinct regulators of immune tolerance, and investigating their role may provide new insights into the pathogenesis of RA. We enrolled 51 RA patients and 27 healthy controls. RA patients were grouped by disease activity: low-to-moderate (L-M RA) or high (H RA). We analyzed clinical data and lymphocyte subsets, including CD4 + Tregs and CD8 + Foxp3+T, to identify key immunological differences between RA and HC. Internal validation was performed using bootstrapping, and P-values were adjusted for false discovery rate (FDR) to ensure robustness. (1) CD4 + Tregs decreased in RA, especially in H RA, while CD8 + Foxp3+T increased, most in L-M RA. (2) CD4 + Tregs correlated negatively with IL-2, IL-4, and IL-10, whereas CD8 + Foxp3+T correlated negatively with ESR, DAS28, and RF-IgA/IgG. (3) The CD4 + Treg/CD8 + Foxp3+T ratio distinguished RA from controls. The combined model (including TNF-α and CD4 + Treg%) demonstrated strong discrimination with an AUC of 0.928 (optimism-corrected AUC: 0.912). CD8 + Foxp3+ T were increased in patients with RA and exhibited a trend opposite to that of CD4 + Treg. These two cell populations were associated with disease activity from distinct immunological perspectives. Furthermore, the CD4 + Treg/CD8 + Foxp3+T ratio and the combination of CD4 + Treg% with TNF-α showed potential value in distinguishing patients with RA from healthy controls. These findings provide additional insight into the immunological characteristics of RA and support further investigation into the biological significance of CD8 + Foxp3+ T cells.

Keywords:  Biomarker; CD4 + regulatory T cells; CD8 + regulatory T cells; Rheumatoid arthritis

DOI:  https://doi.org/10.1007/s10753-026-02543-1