bims-tuinly Biomed News
on Tumor-infiltrating lymphocytes therapy
Issue of 2026–06–28
25 papers selected by
Pierpaolo Ginefra, Ludwig Institute for Cancer Research



  1. J Immunother Cancer. 2026 Jun 25. pii: e015360. [Epub ahead of print]14(6):
       PURPOSE: Treatment strategies for patients with advanced melanoma resistant to adoptive cell therapy with tumor-infiltrating lymphocytes (TILs) remain undefined. The efficacy of immune checkpoint inhibitors (ICIs) after TIL has not been previously described.
    PATIENTS AND METHODS: Patients with unresectable melanoma who received ICI therapy both before and after TIL treatment were retrospectively identified at 14 international centers. Safety was evaluated among all patients that received one dose of ICI therapy after TIL. Objective response rates (ORR) were determined based on investigator-assessed Response Evaluation Criteria in Solid Tumors (RECIST) V.1.1 responses based on radiology reports. Kaplan-Meier methodology was used to estimate overall survival (OS) and melanoma-specific survival.
    RESULTS: Among 133 patients assessed for response to ICI therapy after progression on TIL therapy, the ORR was 11% (n=14; 95% CI 5.9% to 17%), including 3 complete responses (2.3%) and 11 partial responses (8.3%). Among patients treated with a programmed death-1 inhibitor prior to TIL (n=121), the ORR was 8.3% (95% CI 4.0% to 15%). At a median follow-up among survivors of 21 months, median OS from post-TIL ICI was 8.8 months (95% CI 6.5 to 12 months). Among 14 patients with response to post-TIL ICI, 13 received an ICI they had not previously received. In the safety cohort (n=138), post-TIL ICI therapy-related toxicity occurred in 31 patients(22%), among which 7 patients (5.1%) experienced recurrence of an adverse event experienced with prior ICI treatment.
    CONCLUSION: Efficacy with ICI after TIL was limited, with no evidence of synergistic toxicity. As TIL therapy becomes more widely available, these data highlight the need for novel therapies in this setting and support the inclusion of this population in future clinical trials.
    Keywords:  Adoptive cell therapy - ACT; Immune Checkpoint Inhibitor; Skin Cancer
    DOI:  https://doi.org/10.1136/jitc-2026-015360
  2. Cancer Invest. 2026 Jun 21. 1-10
      Nasopharyngeal cancer (NPC) is a major health burden in Indonesia, and reliable prognostic biomarkers are needed to guide treatment decisions. This study examined associations between lymphocyte-monocyte ratio (LMR), tumor-infiltrating lymphocytes (TILs), tumor-associated macrophages (TAMs), and 3-year progression-free survival (PFS) in advanced-stage NPC. A retrospective cohort was assembled at a national referral hematology-oncology clinic, including patients diagnosed from January 2015 to 2020. TIL and TAM expression were assessed by immunohistochemistry, and cutoffs for LMR, CD8, and CD163 were derived using receiver operating characteristic analysis. Kaplan-Meier methods with log-rank testing and Cox proportional hazards regression (univariable and multivariable) were applied. Median 3-year PFS was 24 months (95% CI: 19.44-25.04), with 46.7% event-free at 3 years. High LMR (>1.82) and high CD8+ TIL (>47.5%) were associated with higher 3-year PFS (56.3% and 59.3%) than low groups (31.7% and 30.4%). High CD163+ TAM (≥196) was associated with worse 3-year PFS (9.1% vs 73.8%; p < 0.001). In multivariable models, associations remained significant: LMR aHR 1.844 (p = 0.026), CD8 aHR 2.222 (p = 0.008), and CD163 aHR 5.680 (p < 0.0001). After adjustment for age, ECOG, body mass index, treatment type, sex, and stage (CD163 model), effect estimates were consistent with univariable findings. These biomarkers may aid risk stratification in advanced NPC.
    Keywords:  Lymphocytes; Nasopharyngeal carcinoma; Tumor-associated macrophages; Tumor-infiltrating
    DOI:  https://doi.org/10.1080/07357907.2026.2675655
  3. BMC Cancer. 2026 Jun 22.
       BACKGROUND: Tumor-infiltrating lymphocytes (TILs) are recognized as prognostic biomarkers. However, inconsistent TIL scores across breast cancer stages, and data are scarce for Ethiopian patients. Thus, this study evaluated TIL scores across breast cancer stages in Northwest Ethiopia.
    METHODS: A cross-sectional study was conducted from 20 June 2024 to 19 June 2025. Using consecutive sampling, 62 breast cancer patients were enrolled, and their socio-demographic and clinicopathological data were collected using questionnaires and chart reviews. Formalin-fixed, paraffin-embedded tissue sections were stained with hematoxylin and eosin, and stromal TILs were evaluated according to the International TIL Working Group guidelines. Data were analyzed using SPSS version 27. Median TILs across clinicopathological variables were assessed using Kruskal-Wallis and Mann-Whitney U tests. Spearman correlation assessed the association of TIL scores and clinicopathological variables. P ≤ 0.05 was considered statistically significant.
    RESULTS: Low TIL scores were observed in 76.5% of stage II cases, whereas high TIL scores were observed in 54.5% of stage IV cases. Median TIL percentages in stage IV (48%) and III (33%) were significantly higher than in stage II (7%), and in N2-N3 (33%) than in N0-N1 (9%) (p < 0.001). Moreover, the median TIL percentage in grade III (48%) was higher than in grade I (9%) (p = 0.028). TILs were positively correlated with TNM stage, lymph node stage, and grade.
    CONCLUSIONS: A positive correlation was found between TIL score and TNM stage. Intermediate and high TIL scores were common in advanced stages, whereas low TIL scores were prevalent in early stages. The findings suggest that advanced tumors may harbor higher mutation burdens and immunogenicity. Future research should explore the molecular subtypes of immune profiles.
    Keywords:  Breast cancer; Ethiopia; Tumor-infiltrating lymphocytes
    DOI:  https://doi.org/10.1186/s12885-026-16359-7
  4. Breast Cancer Res Treat. 2026 Jun 26. pii: 60. [Epub ahead of print]217(3):
       INTRODUCTION: Triple-negative breast cancer (TNBC) is an aggressive subtype characterized by poor prognosis. Based on the KEYNOTE-522 trial, neoadjuvant pembrolizumab plus chemotherapy has become the standard of care due to significantly improved pathological complete response (pCR) rates. The presence of tumor-infiltrating lymphocytes (TILs) is a predictive biomarker of pCR. This retrospective cohort study examines a diverse patient population treated with the K522 regimen to determine if TILs predict pCR relative to other clinical and tumor-specific factors.
    METHODS: We retrospectively reviewed 187 patients with early-stage TNBC at two institutions (one tertiary care, one safety-net) who completed neoadjuvant K522 treatment between 2021 and 2024. Statistical analyses included Chi-squared tests, Z-tests, and univariate logistic regression to evaluate associations between TILs, ethnicity, tumor grade, and pCR, and multivariate logistic regression to assess whether the association between dose intensity and pCR differed by the presence of TILs at baseline.
    RESULTS: The overall pCR rate was 57%; TILs were present in 52.8% of cases. TILs were associated with a significantly higher pCR rate (70% vs. 48% without TILs; p = 0.0027). While pCR rates were similar across ethnicities, Hispanic patients with TILs had significantly higher pCR than those without (80.0% vs. 51.5%; p = 0.0254). Controlling for grade, patients with TILs were 2.442 times more likely to achieve pCR (CI: 1.310-4.553; p = 0.0050). Grade 3 tumors and node-positivity with TILs also showed statistically significant rates of pCR. Logistic regression analysis revealed a significant interaction between TIL status and immunotherapy dose intensity on pCR. While dose intensity did not influence pCR in TIL-negative patients, TIL-positive patients achieved higher pCR rates with lower dose intensity (OR = 1.54, p = 0.036).
    CONCLUSION: TILs serve as a strong predictive biomarker for immunotherapy response in a real-world TNBC population. Our findings regarding Hispanic, node-positive patients, and the interaction between TIL status and dose intensity suggest that TILs could guide treatment de-escalation to reduce K522-related toxicity. Standardizing TILs reporting is critical to optimizing treatment strategies and improving outcomes in underrepresented populations.
    Keywords:  Immunotherapy; Neoadjuvant therapy; Pathological complete response; Triple-negative breast cancer; Tumor-infiltrating lymphocytes
    DOI:  https://doi.org/10.1007/s10549-026-08002-7
  5. Biomedicines. 2026 Jun 11. pii: 1325. [Epub ahead of print]14(6):
      Background: Triple-negative breast cancer (TNBC) is a biologically heterogeneous disease. Clinically accessible biomarkers remain limited. Objective: To evaluate androgen receptor (AR) expression and immune response-stromal tumor-infiltrating lymphocytes (sTILs), CD4+, CD8+, CD4/CD8 ratio-to explore their clinicopathological associations and relationships with 3-year overall survival (OS) in TNBC. Methods: We retrospectively analyzed data from 86 treatment-naïve women with TNBC who were treated between 2012 and 2019 at a single academic center. AR and CD4/CD8 were assessed immunohistochemically; sTILs were scored on H&E following The International TIL Working Group recommendations. Survival analyses focused on 3-year OS, with follow-up truncated at 36 months and multivariable Cox regression restricted to non-metastatic disease (stage I-III). Results: High AR expression (≥10%) occurred in 23% of tumors and was associated with lower CD8+ infiltration and lower tumor grade. Across adjusted models, we did not demonstrate statistically significant and independent associations between AR or immune markers and 3-year OS; however, inference is limited by the low number of events (10 deaths). Conclusions: AR status was associated with the immune response, particularly with reduced CD8+ infiltration in AR-high tumors, supporting the concept of biologically distinct AR-immune phenotypes. The absence of statistically significant survival associations in adjusted analyses should be interpreted cautiously given the limited event counts, and larger prospective cohorts are needed to validate prognostic and potential therapeutic implications.
    Keywords:  CD4+; CD8+; androgen receptor (AR); prognosis; triple-negative breast cancer (TNBC); tumor-infiltrating lymphocytes (TILs)
    DOI:  https://doi.org/10.3390/biomedicines14061325
  6. APMIS. 2026 Jun;134(6): e70231
      High levels of tumor-infiltrating lymphocytes (TILs) are associated with improved survival after radiotherapy in breast cancer (BC) patients, particularly in estrogen receptor (ER)-negative disease. This study investigated the prognostic relevance of immune cell subsets in irradiated patients. Node-positive patients (N = 1307) from the Danish Breast Cancer Group internal mammary node (IMN)2 study were included, in which IMN irradiation (IMNI) was allocated by laterality. Tissue microarrays were stained with multiplex immunohistochemistry for CD8+, CD4+, and FOXP3+ T-cells, CD68+ macrophages, and CD11c+ dendritic cells. Digital image analysis quantified immune infiltration across spatial compartments. Prognostic associations with distant recurrence, breast cancer-specific mortality, and overall mortality (OM) were assessed using multivariable flexible parametric survival models, including ER-stratified analyses. T-cell infiltration demonstrated a strong inverse association with OM, with 12-year OM-risk decreasing from 42% in tumors with lowest CD8+ infiltration to 27% in those with highest (HR 0.52, 95% CI (0.36-0.76)). The association was strongest in the ER-negative subgroup. In ER-positive disease, prognostic benefit was observed at low-to-moderate infiltration. CD68+ and CD11c+ infiltration correlated with improved outcomes, but with weaker effects. Immune markers were not predictive of IMNI benefit. These findings demonstrate that T-cell subsets are strong prognostic markers in irradiated BC patients, especially in ER-negative disease.
    Keywords:  breast cancer; digital image analysis; immune cells; radiotherapy; survival
    DOI:  https://doi.org/10.1111/apm.70231
  7. Front Immunol. 2026 ;17 1837084
      Glioblastoma (GBM) represents the most aggressive primary brain tumor in adults, characterized by a profoundly immunosuppressive tumor microenvironment (TME) that systematically disables cytotoxic lymphocyte function and renders conventional immunotherapy largely ineffective. While exhaustion of CD8+ T cells and natural killer (NK) cells within solid tumors has been extensively studied in other cancer types, the CNS-specific architectural, metabolic, and molecular constraints that shape cytotoxic lymphocyte heterogeneity in GBM remain insufficiently characterized. Recent advances in single-cell RNA sequencing (scRNA-seq) and spatial multiomics have begun to reveal a rich landscape of cytotoxic lymphocyte subpopulations in GBM. These include TCF-1+ progenitor-exhausted T cells (Tpex), terminally exhausted CD8+ T cells (Tex), and dysfunctional natural killer (NK) cell subsets, each distributed across anatomically distinct immune niches. This review synthesizes current knowledge across three interconnected areas: the single-cell atlas of GBM-infiltrating cytotoxic lymphocytes; the spatial organization of their dysfunction within perinecrotic, perivascular, and infiltrative-edge niches; and the epigenetic and transcriptional programs that underlie GBM-specific cytotoxic failure, including dysregulation of the TOX/TCF-1 axis and IDH-mutation-driven silencing of NKG2D ligands. Critically, we compare CD8+ T cell and NK cell exhaustion mechanisms, highlighting their mechanistic divergence and therapeutic implications. We further discuss how these multiomics insights can be translated into neurosurgically relevant strategies, including intraoperative tumor profiling, progenitor T cell expansion via epigenetic priming, NKG2A/TIGIT dual blockade, and intracavitary delivery of engineered NK cells. Together, this review proposes a spatially and cellularly resolved framework for understanding cytotoxic immune failure in GBM and outlines precision immunotherapy approaches tailored to the unique immunobiology of the CNS tumor microenvironment.
    Keywords:  NK cell dysfunction; blood-brain barrier; cytotoxic lymphocytes; glioblastoma; immunotherapy; progenitor exhausted T cells; single-cell transcriptomics; spatial genomics
    DOI:  https://doi.org/10.3389/fimmu.2026.1837084
  8. Front Immunol. 2026 ;17 1877547
       Background: Early identification of breast cancer patients unlikely to benefit from neoadjuvant therapy (NAT) remains a critical unmet need. This study aimed to develop and internally validate a multimodal prediction model for NAT non-response by integrating clinicopathological, tumor microenvironment (TME), longitudinal magnetic resonance imaging (MRI), and systemic inflammatory features.
    Methods: In this retrospective study, 112 patients with primary breast cancer underwent baseline MRI, a second MRI after two NAT cycles, and definitive surgery. Non-response was defined as Miller-Payne grades 1-3. Candidate predictors were categorized into four domains. After univariate screening, domain-specific multivariable logistic regression was performed, and retained variables entered least absolute shrinkage and selection operator (LASSO) regression to construct a final multimodal model. Internal validation included five-fold cross-validation and 500-iteration bootstrap. Calibration and decision curve analyses were also performed.
    Results: Thirty-eight patients (33.9%) were non-responders. The individual domain models achieved apparent AUCs of 0.844 (clinical), 0.786 (imaging), 0.828 (TME), and 0.706 (inflammatory). Following LASSO selection, nine features were retained: HER2 status, ER status, Ki-67 index, late enhancement rate after two cycles (LER2), baseline background parenchymal enhancement (BPE), time to peak after two cycles (TTP2), tumor-stroma ratio (TSR), tumor-infiltrating lymphocytes (TILs), and pan-immune-inflammation value after two cycles (PIV2). The multimodal model yielded an apparent AUC of 0.933 (95% CI: 0.890-0.977), with a bootstrap-corrected AUC of 0.855 and a mean five-fold cross-validation AUC of 0.908 ± 0.038. TILs, TSR, PIV2, and Ki-67 were independent predictors. The model demonstrated acceptable calibration after correction for optimism and a net clinical benefit across a range of thresholds.
    Conclusions: A multimodal prediction model integrating clinicopathological, imaging, tumor microenvironment, and systemic inflammatory features showed potential for early identification of breast cancer patients unlikely to benefit from neoadjuvant therapy. However, given the limited sample size and exploratory single-center design, performance estimates should be interpreted cautiously, and external validation is essential.
    Keywords:  breast cancer; magnetic resonance imaging; neoadjuvant therapy; non−response; pan−immune−inflammation value; tumor microenvironment; tumor−infiltrating lymphocytes
    DOI:  https://doi.org/10.3389/fimmu.2026.1877547
  9. Res Sq. 2026 Jun 11. pii: rs.3.rs-9929244. [Epub ahead of print]
      CD8⁺ T cells mediate host defense and tumor immunity through specialized differentiation states, yet the regulatory programs that guide these states may also limit their functional potential. Loss-of-function studies have defined many regulators required for T cell differentiation, but they do not readily reveal regulatory activities that emerge only when transcription factors are ectopically expressed outside their native lineage, dosage, or temporal context. Here, we developed single-cell gain-of-functon(GOF) sequencing (scGOF-seq), a multiplexed platform for in vivo mapping of transcription factor overexpression in antigen-specific CD8⁺ T cells across immunocompetent models of infection and cancer. By enforcing expression of canonical T cell regulators, lineage-silenced developmental factors, and temporally restricted transcription factors, scGOF-seq uncovered unexpected in vivo activities. Developmental regulators normally silenced in T cells, including NANOG, SOX2, OCT4 and GATA2, reshaped T cell differentiation in context-dependent ways, with NANOG promoting stemness-associated phenotypes and accumulation during chronic infection. In parallel, sustained cMyc expression outside its native temporal window generated a stem-like, effector-featured state with enhanced metabolic fitness, reduced terminal exhaustion, and profound antigen-dependent expansion exceeding 5,000-fold. Importantly, cMyc GOF maintained cell-cycle checkpoint signatures and demonstrated a strong dependence on antigen presence for proliferation across the tested conditions. scGOF-seq further identified cooperating transcription factor modules that complemented cMyc-driven programs and improved T cell responses in solid tumors. These findings establish systematic GOF perturbation as a framework for uncovering latent and temporally constrained regulatory activities in CD8⁺ T cells and guiding immune-state engineering.
    DOI:  https://doi.org/10.21203/rs.3.rs-9929244/v1
  10. Nat Commun. 2026 Jun 24.
    GAMBIT Consortium
      Pathologic complete response after neoadjuvant treatment is considered a surrogate of cure in triple-negative breast cancer, yet around 10% of patients still relapse. Whether baseline stromal tumor-infiltrating lymphocytes can stratify this residual risk is unknown. Here, we report on GAMBIT, a multicentric real-world retrospective study of 2457 patients with triple-negative breast cancer or estrogen receptor-low disease, HER2-negative, of whom 1192 obtained a pathological complete response and 690 have evaluable tumor infiltrating lymphocytes. Among patients with pathological complete response, clinical nodal status and tumor infiltrating lymphocytes are independently prognostic and patients with clinical node-positive/low-tumor infiltrating lymphocytes tumors experience substantially worse outcomes, with five-year distant relapse-free survival of 83.4% and overall survival of 85.8%. In this high-risk subgroup, five-year cumulative incidence of central nervous system reaches 7.5%, including 6.9% presenting as isolated central nervous system relapse. In this work, we identify a high-risk subgroup despite pathologic complete response and provide a framework supporting risk-adapted trial design incorporating central nervous system-directed strategies.
    DOI:  https://doi.org/10.1038/s41467-026-74056-2
  11. Surg Case Rep. 2026 ;pii: 26-0180. [Epub ahead of print]12(1):
       INTRODUCTION: Spontaneous regression (SR) of colorectal cancer (CRC) is exceptionally rare. The SR of malignant tumors occurs in approximately 1 in 80000-100000 cases, and CRC accounts for <2% of all spontaneous malignancy regressions. Recent studies have suggested that immunological mechanisms, particularly those related to deficient mismatch repair (dMMR) and high-frequency microsatellite instability, may play an important role in such tumor regressions.
    CASE PRESENTATION: A 68-year-old woman was referred to our hospital after a positive fecal occult blood test. Colonoscopy revealed a 12-mm nonpolypoid lesion (IIa + IIc) in the transverse colon. Biopsy specimens showed poorly differentiated carcinoma without glandular formation or mucin production, accompanied by marked tumor-infiltrating lymphocytes (TILs). Immunohistochemistry was negative for CK20, CDX2, and neuroendocrine markers. The majority of TILs were CD3-positive lymphocytes. CT revealed no lymph node involvement or distant metastasis (cT1bN0M0). Laparoscopic partial colectomy of the transverse colon with D3 lymphadenectomy was performed 50 days after biopsy. Macroscopically, the resected specimen showed only a small scar-like lesion, and histological examination revealed no residual carcinoma. All dissected lymph nodes were tumor-free. Additional immunohistochemical analysis of the biopsy specimen showed loss of MLH1 and PMS2 expression, consistent with a dMMR status. These findings highlighted the possibility of medullary carcinoma, but a definitive diagnosis was not possible due to the limited biopsy samples. The postoperative course was uneventful, and no recurrence was observed during the 18 months of follow-up period without adjuvant therapy.
    CONCLUSIONS: We report an extremely rare case of SR of poorly differentiated CRC with dMMR and marked TILs. Enhanced tumor immunogenicity associated with dMMR and immune activation may contribute to CRC regression.
    Keywords:  deficient mismatch repair; high-frequency microsatellite instability; spontaneous regression; tumor-infiltrating lymphocytes
    DOI:  https://doi.org/10.70352/scrj.cr.26-0180
  12. Cancers (Basel). 2026 Jun 10. pii: 1887. [Epub ahead of print]18(12):
      Background: Buparlisib, combined with paclitaxel, improved survival in BERIL-1 trial patients with recurrent/metastatic head and neck squamous cell carcinoma (R/M HNSCC). However, predictive biomarkers of benefit remain undefined. Objective: To evaluate whether spatial biomarkers extracted from hematoxylin and eosin (H&E) slides using artificial intelligence (AI) can predict overall survival benefit from buparlisib. Methods: Whole-slide H&E images from BERIL-1 trial patients were analyzed using a deep learning model trained to segment tissue compartments and classify cell phenotypes. Three predefined spatial features were evaluated: tumor-infiltrating lymphocyte density, tumor microenvironment heterogeneity, and granulocyte fraction in the tumor invasive margin. Cox proportional hazards model assessed biomarker-treatment interactions. Results: Of 158 trial participants, 144 had available slides. High tumor-infiltrating lymphocyte density (>10%) was associated with significantly improved overall survival with buparlisib versus placebo (HR, 0.25 (95% CI, 0.01-0.64; p = 0.002)), as were high tumor microenvironment heterogeneity (HR, 0.47 (95% CI, 0.27-0.80; p = 0.005)) and granulocyte enrichment in the tumor invasive margin (HR, 0.51 (95% CI, 0.30-0.88; p = 0.01)); within-arm proximity analysis showed higher granulocyte-tumor-cell proximity correlated with improved overall survival on buparlisib (HR, 0.32 (95% CI, 0.18-0.58; p < 0.001)). AI-derived spatial metrics outperformed CD3 immunohistochemistry. Among oropharyngeal tumors, HPV-positive cases were more frequent in patients with high tumor-infiltrating lymphocytes. Conclusions: AI-extracted spatial features from H&E slides were associated with overall survival benefit from buparlisib in R/M HNSCC. These scalable biomarkers support image-based patient selection strategies and are being prospectively evaluated in the BURAN phase 3 trial.
    Keywords:  BERIL-1; H&E; artificial intelligence; buparlisib; digital pathology; head and neck cancer; spatial biomarkers; squamous cell carcinoma
    DOI:  https://doi.org/10.3390/cancers18121887
  13. Front Immunol. 2026 ;17 1865742
      Osteosarcoma remains a major treatment challenge, especially for patients with recurrent, refractory or metastatic diseases. Adoptive cell therapy (ACT), including CAR-T cells, TCR engineered T cells, CAR-NK cells and macrophage based cell therapy, provides a promising strategy for redirecting immune effector cells to fight osteosarcoma. However, clinical translation has been limited by antigen heterogeneity, on-target/off-tumor toxicity, insufficient tumor trafficking, poor persistence, functional exhaustion, and the immunosuppressive tumor microenvironment. This mini review discusses emerging innovations designed to overcome these safety and efficacy barriers. First, engineering strategies such as multi-antigen recognition, logic-gated CAR systems, suicide switches, transient CAR expression, armored cytokine circuits, checkpoint-resistant designs, and chemokine receptor modification may improve precision, controllability, and durability. Second, vaccination approaches may serve as programmable amplifiers of ACT by promoting in vivo expansion, immune memory, antigen spreading, and local inflammatory priming. Third, tumor microenvironment remodeling through stromal modulation, vascular normalization, myeloid reprogramming, checkpoint blockade, and metabolic intervention may convert osteosarcoma into a more permissive niche for cellular therapy. Collectively, next-generation ACT for osteosarcoma will likely require modular, biomarker-guided combinations that integrate cellular engineering, vaccine-based boosting, and microenvironmental remodeling to achieve safer and more durable antitumor responses.
    Keywords:  CAR-T cells; adoptive cell therapy; antigen heterogeneity; cellular engineering; immunotherapy; osteosarcoma; tumor microenvironment; vaccination
    DOI:  https://doi.org/10.3389/fimmu.2026.1865742
  14. Int Immunopharmacol. 2026 Jun 24. pii: S1567-5769(26)00912-4. [Epub ahead of print]186 117066
      Lactate accumulation and the acidic tumor microenvironment play pivotal roles in tumor immune evasion, and regulatory T cells (Tregs) constitute a major barrier to effective anti-tumor immunity. However, whether lactate or the acidic microenvironment can drive the generation of Tregs, and the underlying mechanisms involved, remain poorly understood. Here, we demonstrate that the expression of FOXP3 is positively correlated with the expression of LDHA across multiple tumor types. Lactic acid, but not sodium lactate, enhanced the expression of FOXP3 by activating the latent TGF-β1, which derived from the culture medium containing fetal bovine serum or produced by tumor cells. Knockdown of LDHA in tumor cells reduced the production of lactic acid and significantly delayed tumor growth in immunocompetent mice, accompanied by a decrease in Tregs and an enhancement of anti-tumor function of T cells. Our findings reveal a novel mechanism by which lactic acid facilitates the generation of Tregs and promotes cancer progression, and further suggest that targeting lactic acid metabolism or the acidic tumor microenvironment may represent a promising therapeutic strategy.
    Keywords:  Antitumor immunity; FOXP3; LDHA; Lactic acid; Latent TGF-beta 1; Regulatory T cells
    DOI:  https://doi.org/10.1016/j.intimp.2026.117066
  15. J Transl Med. 2026 Jun 22.
       BACKGROUND: Chemoimmunotherapy has become the standard first-line treatment for advanced non-small cell lung cancer (NSCLC). Deciphering the T-cell subset responsible for chemoimmunotherapy and easily tested conveniently is critical in predicting the treatment outcomes.
    METHODS: Based on peripheral blood collected from patients enrolled from a phase 2 clinical study (ClinicalTrials.gov NCT04836728), we performed multi-color flow cytometry and unsupervised analysis to explore correlations with therapeutic outcomes. We integrated single-cell RNA and T-cell receptor (TCR) sequencing in 36 samples, including peripheral blood, tumors and non-tumor tissues, from 8 NSCLC patients to interpret the correlation, which was further verified using blood samples, orthotopic and subcutaneous lung cancer mouse model.
    RESULTS: The baseline CD28-KLRG1+CD57+ and on-treatment CD28-KLRG1+ CD8+ T cells in peripheral blood were independent factors which indicated improved treatment outcomes in advanced NSCLC patients receiving first-line chemoimmunotherapy. While being in a late-differentiated T-cell status, these cells were clonally expanded and reinvigorated during chemoimmunotherapy, serving as a peripheral T-cell pool for supplying potential tumor-reactive T cells in tumors, and reversely differentiating into less-differentiated subsets. The zinc-metallothionein pathway regulated the CD28-KLRG1+ CD8+ T-cell subset. Zinc supplementation combined with chemoimmunotherapy improved both local and systemic antitumor immune responses in mouse model.
    CONCLUSIONS: Circulating CD28-KLRG1+ CD8+ T cells are valuable and convenient biomarkers for first-line chemoimmunotherapy in advanced NSCLC and provide insight into how late-differentiated or senescent T cells engage in the antitumor immunity when immunotherapy is added to conventional therapies.
    Keywords:  CD8+ T cell; Immunotherapy; Metallothionein; Non-small cell lung cancer; Tumor-infiltrating T cell
    DOI:  https://doi.org/10.1186/s12967-026-08484-5
  16. Sci Rep. 2026 Jun 21.
      Double-negative T cells (DNTs; CD3+CD4-CD8-) have been implicated in immune regulation in autoimmune settings, but their relevance to reproductive tissue immune balance and inflammation-associated infertility remains unclear. Here, we define a population of TCRβ+NK1.1- DNTs enriched in the mouse ovary and uterus and profiled its phenotype, tissue behavior, and function in inflammatory models associated with impaired fertility. In parallel, we performed RNA-seq on spleen- and thymus-derived NK1.1- DNT and CD8+ T cell enriched population. Relative to CD8+ T cells, peripheral NK1.1- DN T-cell-enriched populations displayed an activated, regulatory-like transcriptional profile with reduced Cd8a/Cd8b1, Il7r, and cytotoxic effector markers, alongside increased expression of Pdcd1, Lag3, Tox, and Il10. Ex vivo, FACS-sorted splenic DNTs produced low inflammatory cytokine output after CD3/CD28 stimulation and suppressed CD8+ T-cell proliferation primarily through contact-dependent mechanisms. In vivo, ovarian DNTs decreased following CD8-targeting depletion, and ovarian and uterine DNTs showed limited exchange in parabiosis. In chronic interferon-γ-driven inflammation (ARE-/-) and zona pellucida 3-induced ovarian inflammation, reproductive tissues exhibited reduced DNT frequencies, a shift in the DNT: CD8+T ratio favoring CD8+ T cells and increased activated CD8+ phenotypes. Finally, adoptive transfer of ex vivo FACS-sorted wild-type DNTs into ARE-/- females increased pregnancy frequency, supporting DNT-associated immunoregulation as a feature of inflammation-associated infertility.
    Keywords:  CD8+ T cells; Double-negative T cells; IFN-γ-mediated inflammation; Ovarian immune tolerance; Uterine immune tolerance
    DOI:  https://doi.org/10.1038/s41598-026-57959-4
  17. World J Surg Oncol. 2026 Jun 25.
       PURPOSE: To develop a fatty acid metabolism-based deep learning model for predicting biochemical recurrence (BCR) in prostate cancer (PCa) and to identify recurrence-associated metabolic regulators.
    METHODS: Transcriptomic data from TCGA and GEO GSE70769 were integrated to identify fatty acid metabolism-related genes and construct a deep learning model for BCR prediction. Tumor-infiltrating lymphocytes (TILs) were quantified from H&E-stained slides using a convolutional neural network-based approach. Single-cell RNA sequencing data were analyzed to identify candidate metabolic regulators enriched in malignant epithelial cells. Immunohistochemistry was performed to examine the protein expression patterns of NUDT19 and its expression associations with key fatty acid metabolism enzymes, including FASN, ACACA, and CPT1A. Functional roles were further evaluated using in vitro assays, xenograft models, and serum metabolomics.
    RESULTS: The model effectively stratified PCa patients into high- and low-risk groups with distinct BCR-free survival outcomes. The high-risk group showed increased TIL infiltration, suggesting a more immune-infiltrated or inflammatory tumor microenvironment. Integrated single-cell and bulk transcriptomic analyses identified NUDT19 as a candidate metabolic regulator predominantly expressed in malignant epithelial cells and positively correlated with FASN, ACACA, and CPT1A. NUDT19 knockdown suppressed PCa cell proliferation, migration, and invasion, induced apoptosis, and inhibited tumor growth in vivo. Serum metabolomics further revealed that differential metabolites were enriched in fatty acid metabolism-related pathways.
    CONCLUSIONS: This study establishes a fatty acid metabolism-based deep learning model for BCR prediction and identifies NUDT19 as a candidate metabolic regulator associated with lipid metabolic remodeling and PCa progression. These findings suggest a metabolically active, inflammation-associated recurrence subtype and support further investigation of NUDT19 as a potential therapeutic target in PCa.
    Keywords:  Biochemical recurrence; Deep learning; Fatty acid metabolism; NUDT19; Prostate cancer
    DOI:  https://doi.org/10.1186/s12957-026-04446-9
  18. Pharmaceuticals (Basel). 2026 Jun 09. pii: 913. [Epub ahead of print]19(6):
      Background: Cutaneous melanoma (CM) is a highly immunogenic malignant neoplasm. It features high mutational burden and intense lymphocytic infiltration, supporting the use of immunotherapies, especially inhibitors of the programmed cell death protein 1 (PD-1) checkpoint. Despite advances with anti-PD-1 therapies, such as nivolumab and pembrolizumab, many patients still experience resistance. This result highlights additional immunosuppressive mechanisms within the tumor microenvironment (TME) that limit T-lymphocyte-mediated responses. Objectives: The aim was to discuss the immunologic and metabolic bases of PD-1- and CD73-mediated pathways and evidence that CD73 inhibition can boost PD-1 inhibitor efficacy by acting on convergent immunosuppressive pathways. Methods: We conducted a narrative literature review focusing on tumor immunosuppression, purinergic signaling and checkpoint inhibitor-based immunotherapy. Results: The purinergic pathway, mediated by the ectonucleotidase CD73, is a critical regulator of tumor immunosuppression. CD73 converts extracellular adenosine monophosphate (AMP) into adenosine. This adenosine accumulates in the hypoxic and inflamed TME, exerting immunosuppressive effects. Adenosine acts as a "metabolic brake," inhibiting proliferation, cytokine production, and cytotoxic activity of CD8+ T lymphocytes and natural killer (NK) cells. It also promotes the expansion of regulatory T cells (Tregs) and tumor progression. This axis may limit responses to PD-1 blockade, suggesting that complementary pathways are active. Conclusions: Integration of PD-1 and CD73 pathways suggests that CD73 inhibition may enhance PD-1 blockade by targeting convergent immunosuppressive mechanisms. This supports the exploration of combination strategies to broaden the benefits of immunotherapy in CM.
    Keywords:  immune checkpoint inhibitors; immunosuppression; melanoma; purinergic signaling; resistance to immunotherapy; tumor microenvironment
    DOI:  https://doi.org/10.3390/ph19060913
  19. Int Rev Immunol. 2026 Jun 27. 1-18
      Metabolic reprogramming within the tumor microenvironment (TME) is a pivotal driver of CD8+ T cell dysfunction in cancer. Tumor cells outcompete T cells for essential nutrients, including glucose and amino acids, while accumulating immunosuppressive metabolites such as lactate and 2-hydroxyglutarate. Beyond direct functional impairment, emerging research reveals that these metabolic alterations orchestrate CD8+ T cell transcriptional programs by remodeling their epigenome-via histone modifications, DNA methylation, and non-coding RNA networks-thereby dictating their differentiation, cytotoxic potential, and memory formation. A deeper understanding of how TME-derived metabolic signals shape the epigenetic landscape of CD8+ T cells is crucial for improving current cancer immunotherapeutic strategies. This review systematically delineates how key TME metabolic features, including nutrient deprivation and oncometabolite accumulation, regulate CD8+ T cell fate through epigenetic pathways. Furthermore, we discuss promising therapeutic strategies that target the metabolism-epigenetics axis to reinvigorate CD8+ T cell anti-tumor immunity, offering novel perspectives for enhancing adoptive cell therapy and immune checkpoint blockade.
    Keywords:  CD8+ T cells; Cancer immunotherapy; epigenetics; metabolic reprogramming; tumor microenvironment
    DOI:  https://doi.org/10.1080/08830185.2026.2687543
  20. J Immunother Cancer. 2026 Jun 25. pii: e014940. [Epub ahead of print]14(6):
       BACKGROUND: Understanding resistance to anti-programmed cell death protein 1 (PD-1) therapy is critical for developing reversal strategies. Although ectonucleotidase CD39 (ENTPD1) (CD39)+ cluster of differentiation 8 (CD8)+ T cells have been associated with antitumor responses, the naïve regional distribution of these cells and their dysregulated dynamics as determinants of response or resistance to anti-PD-1 monoclonal antibody (mAb) therapy remain poorly characterized.
    METHODS: We employed targeted biopsy techniques to characterize the spatial infiltration of CD39+CD8+ T cells within the tumor microenvironment (TME) of non-small cell lung cancer (NSCLC). It was further performed independent NSCLC cohort, flow cytometry, multiplex immunohistochemistry, single-cell RNA sequencing and functional studies to investigate these cells in vitro and in vivo.
    RESULTS: Region-specific CD39+CD8+ T cells located outside hypoxic zones initially served as potential biomarkers to predict response to anti-PD-1 therapy. However, hypoxia gradients within the TME were associated with regional depletion of these cells in NSCLC. Subsequently, anti-PD-1 mAb treatment further reduced intratumoral CD39+CD8+ T-cell levels, even in the context of improved oxygenation following tumor regression, suggesting a feedback mechanism modulating antitumor immunity. Multiple assays revealed that the absence of CD39 impaired functional restoration of CD8+ T cells on PD-1 blockade. This finding was corroborated by distinct transcriptional profiles, enhanced cytotoxic/effector signatures, and a more oligoclonal and expanded T-cell receptor (TCR) repertoire in CD39+CD8+ T cells across the CD8+ T-cell differentiation continuum.
    CONCLUSIONS: These findings identify CD39+CD8+ T cells as an important regulator of anti-PD-1 therapy, whereas initial hypoxia followed by anti-PD-1 treatment sequentially shapes tumor-regional localization of these cells, ultimately linking to suboptimal immunotherapeutic responses.
    Keywords:  Immunotherapy; Lung Cancer; T cell; Tumor microenvironment - TME
    DOI:  https://doi.org/10.1136/jitc-2026-014940
  21. Cureus. 2026 May;18(5): e109160
       BACKGROUND: Neoadjuvant chemotherapy (NAC) is the standard of care for biologically aggressive early breast cancer, and pathological complete response (pCR) is a robust surrogate of long-term outcome. Real-world cohorts integrating clinical, pathological, and patient-reported variables remain limited.
    METHODS: We retrospectively analyzed 149 consecutive women with stage I-III invasive breast cancer treated with NAC followed by curative surgery at Memorial Hospital, Bucharest, Romania. The primary endpoint was overall pathological complete response (pCR), defined as the absence of residual invasive disease in both breast and axillary lymph nodes (ypT0/Tis ypN0; residual ductal carcinoma in situ permitted). Clinicopathological variables, NAC regimen, residual cancer burden (RCB), stromal tumor-infiltrating lymphocytes (TILs), and patient-reported outcomes (European Organisation for Research and Treatment of Cancer {EORTC} global health, Hospital Anxiety and Depression Scale {HADS}) at baseline and end of NAC were analyzed. Predictors of pCR were identified by univariate analysis and multivariable logistic regression with internal bootstrap validation.
    RESULTS: Median age was 54 years (IQR: 46-62). Subtype distribution was luminal B 68/149 (45.6%), human epidermal growth factor receptor 2 (HER2)-positive 49/149 (32.9%), and triple-negative 32/149 (21.5%). Overall, pCR was achieved in 41/149 patients (27.5%) as follows: 6/68 (8.8%) in luminal B, 26/49 (53.1%) in HER2-positive, and 9/32 (28.1%) in triple-negative breast cancer (TNBC) (χ²=27.95, df=2, p<0.001). pCR rates also differed markedly across NAC regimens (χ²=27.55, df=6, p<0.001), ranging from 1/17 (5.9%) for TC to 12/19 (63.2%) for taxane/trastuzumab/pertuzumab (THP). On multivariable analysis, clinical subtype, stromal TILs, and clinical nodal status were the principal independent predictors of pCR; the integrated model achieved an area under the receiver operating characteristic curve (AUC) of 0.93 (bootstrap-corrected: 0.92), with adequate calibration (Hosmer-Lemeshow χ²=4.90, df=6, p=0.557). EORTC global health declined by a median of four points between baseline and end of NAC (Wilcoxon W=2615.5, p<0.001), with 51/149 patients (34.2%) experiencing a clinically meaningful decline. At baseline, 30/149 patients (20.1%) had clinically significant anxiety, and 57/149 (38.3%) had borderline depressive symptoms; clinical depression emerged de novo in 18/149 (12.1%) by the end of NAC. Baseline patient-reported outcomes did not significantly predict pCR after adjustment for biological variables (likelihood-ratio χ²=4.71, df=3, p=0.194).
    CONCLUSIONS: In this Romanian single-center cohort, pCR was independently predicted by clinical subtype and a limited set of biological variables, in line with contemporary literature. Although baseline patient-reported outcomes did not improve prediction of pathological response, the high prevalence of pre-treatment anxiety and the emergence of clinical depression during NAC argue for systematic psychosocial screening as part of standard neoadjuvant care.
    Keywords:  breast cancer; hads; neoadjuvant chemotherapy; pathological complete response; patient-reported outcomes; quality of life; real-world data; tumor-infiltrating lymphocytes
    DOI:  https://doi.org/10.7759/cureus.109160
  22. Cells. 2026 Jun 19. pii: 1115. [Epub ahead of print]15(12):
      Background: The tumor immune microenvironment, particularly the role of cytotoxic CD8+ T lymphocytes, is crucial in cancer progression but remains poorly understood in pituitary neuroendocrine tumors (PitNETs). The significance of CD8+ cell infiltration varies across PitNET subtypes, suggesting a complex interplay with tumor cell lineage. This study aimed to characterize the distribution of CD8+ tumor-infiltrating lymphocytes across different PitNET subtypes defined by the current WHO classification and to explore their association with clinicopathological features. Methods: We conducted a retrospective study on 40 surgically resected PitNETs. All cases were classified based on immunohistochemical expression of pituitary hormones and lineage-specific transcription factors (PIT-1, TPIT, SF-1). CD8+ lymphocyte density was quantified using immunohistochemistry and calculated as cells/mm2. Exploratory statistical analysis was performed based on non-parametric tests to compare CD8+ cell density across tumor subtypes and with parameters like tumor size, invasiveness (Knosp grade), and proliferation index (Ki-67). Findings are to be treated as observational trends. Results: The highest density of CD8+ lymphocytes was observed in plurihormonal PIT-1-positive tumors [17.61 cells/mm2 (IQR: 17.61-60.36)], followed by somatotroph [13.2 (6.6-15.72)] and mammosomatotroph [13.83 (0-21.38)] tumors. A difference in CD8+ density was found between PIT-1-positive and PIT-1-negative tumors (n1 = 34, n2 = 6, U = 49.5, pexact = 0.050, r = 0.33); the medium effect size indicates a possible lineage-related trend. Another difference was observed between SF-1-positive and SF-1-negative tumors (p = 0.025), with SF-1 lineage tumors showing the lowest infiltration. No correlations were found between CD8+ density and tumor size, Knosp grade, or Ki-67 index. Conclusions: The distribution of intratumoral CD8+ T lymphocytes in PitNETs is highly heterogeneous and appears to be strongly dictated by the transcription factor-defined tumor lineage rather than by traditional clinicopathological markers of aggressiveness. PIT-1 lineage tumors harbor a more active immune microenvironment, while SF-1 lineage tumors are relatively 'immune-poor'. These findings highlight the immunological diversity of PitNETs and support further investigation of the tumor immune landscape. Collaborative multi-institutional studies are required to validate these trends.
    Keywords:  CD8+ lymphocytes; PitNETs; immune microenvironment; transcription factors
    DOI:  https://doi.org/10.3390/cells15121115
  23. Curr Med Res Opin. 2026 Jun 21. 1-13
       BACKGROUND: Globally, lung cancer is the most common malignancy among males, with higher incidence and mortality rates than in females. Sex chromosome analyses may provide insights into these disparities. KDM5D, located on the Y chromosome, is recognized as a tumor suppressor. Ongoing studies explore KDM5D's role in malignancies and therapeutic pathways. We evaluated the association of immunohistochemical expression with clinicopathologic parameters and survival.
    METHODS: A total of 102 male lung carcinoma patients who underwent trucut or resection procedures between June 1, 2014, and October 31, 2023, at a tertiary care center were analyzed. Evaluated parameters included age, tumor type, TNM stage, tumor-infiltrating lymphocytes, and KDM5D expression. Additional parameters were also assessed for resections. The JAD1D (KDM5D) antibody was applied to unstained slides, expression levels were quantified using the Q score method, and a cut-off was determined by the ROC curve.
    RESULTS: A cut-off value of 8 was calculated for mortality, but no significant value was found for recurrence or progression. Patients with high expression (>8) had significantly lower overall survival. Higher levels were associated with increased maximum tumor diameter, involvement of pN1, M1 and above, stage IVA-IVB disease, and mild to moderate tumor-infiltrating lymphocytes. These features and elevated KDM5D were significantly higher in the exitus group.
    CONCLUSION: KDM5D expression showed an unadjusted association with mortality in univariate Cox regression. However, this association was not significant after adjusting for clinicopathological variables or correcting for multiple comparisons. Given conflicting findings in the literature, further studies, methodological standardization, and deeper exploration of KDM5D and the KDM5 family are warranted.
    Keywords:  KDM5D; Lung; carcinoma; pathology; survival
    DOI:  https://doi.org/10.1080/03007995.2026.2689251
  24. Front Immunol. 2026 ;17 1840895
      The shift toward precision medicine in cancer immunotherapy has increased demand for rapid, scalable production platforms for personalized vaccine antigens targeting neoantigens and tumor-associated antigens. Escherichia coli-based recombinant protein production represents a globally established system offering speed and cost-effectiveness, yet the immunogenic potential of soluble antigens produced via this platform remains incompletely characterized. Here, we systematically evaluated the capacity of E. coli-derived soluble antigen formulations to elicit anti-tumor T cell responses. Using ClearColi BL21 (DE3), a lipopolysaccharide (LPS)-truncated strain free of endotoxic contamination, we produced soluble formulations containing murine OVA-derived epitopes. In vitro antigen presentation assays revealed minimal T cell activation despite high numbers of CD8+ and CD4+ T cells, indicating compromised antigen presentation capacity. However, in vivo adoptive transfer experiments demonstrated strongly inducible T cell expansion in draining and non-draining lymph nodes, with high frequencies of antigen-specific CD8+ T cells producing IFN-γ and TNF-α. Prime-boost vaccination strategies in experimental melanoma models achieved protective efficacy and durable survival with persistent antigen-specific T cell responses throughout the observation period. Under stringent single-prime conditions, transient T cell frequencies were observed without long-term tumor control.
    Keywords:  E. coli expression system; adoptive transfer; cancer vaccines; immunotherapy; multi-epitope antigens; protein-based vaccine; therapeutic strategies
    DOI:  https://doi.org/10.3389/fimmu.2026.1840895
  25. Cancer Res. 2026 Jun 22.
      Cancer treatment using immune checkpoint blockade (ICB) with anti-PD-1 and anti-CTLA-4 has been successful. However, primary and acquired resistance limits clinical benefit. To improve the effectiveness of ICB therapies, strategies that reorchestrate anti-tumor immunity through mechanism-based drug combinations are being actively explored. The alkylating chemotherapeutic agent cyclophosphamide (CTX) has direct tumoricidal and immunomodulatory properties, including the induction of homeostatic proliferation of T cells. Since ICB suppresses inhibitory signals in T cells, ICB might be able to augment CTX-induced homeostatic proliferation of antigen-specific T cells, thereby resetting the T cell receptor (TCR) repertoire in favor of tumor-specific T cells. Here, we showed that a single dose of CTX one day prior to starting αPD-1+αCTLA-4 treatment was sufficient to delay tumor progression in established melanoma and prolong survival in tumor-bearing mouse models. These effects extended to other lymphodepleting treatments, such as gemcitabine and radiation therapy. The anti-tumor immune response was mainly driven by the clonal expansion of activated/effector CD8+ tumor infiltrating lymphocytes. Furthermore, combined CTX and αPD-1+αCTLA-4 treatment demonstrated efficacy across additional preclinical tumor models, including colorectal cancer and triple negative breast cancer. Overall, these findings highlight that the combination of CTX and ICB represents a clinically relevant approach in the treatment of immunotherapy-refractory tumors.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-25-3863