Journal Articles

Time-of-day of first checkpoint inhibitor dose influences clinical outcomes and immune responses in hepatocellular carcinoma Spotlight 

Among a retrospective cohort of 84 HCC patients treated with ICB, those who received their first ICB dose in the morning (prior to 12 noon) had increased PFS (and a trend in OS) compared to those receiving a first dose in the afternoon. The timing of subsequent doses did not have a similar stratifying effect, and morning dosing did not raise the rate of irAEs. Comparing baseline and early on-treatment blood samples, Li et al. found that patients first receiving ICB in the morning had diminished induction of certain cytokines (IL-6, IL-1B, VEGF-A, and IL-21) and a greater expansion of cytotoxic CD8+ Tcm cells, compared to those receiving an afternoon dose.

Contributed by Alex Najibi

Among a retrospective cohort of 84 HCC patients treated with ICB, those who received their first ICB dose in the morning (prior to 12 noon) had increased PFS (and a trend in OS) compared to those receiving a first dose in the afternoon. The timing of subsequent doses did not have a similar stratifying effect, and morning dosing did not raise the rate of irAEs. Comparing baseline and early on-treatment blood samples, Li et al. found that patients first receiving ICB in the morning had diminished induction of certain cytokines (IL-6, IL-1B, VEGF-A, and IL-21) and a greater expansion of cytotoxic CD8+ Tcm cells, compared to those receiving an afternoon dose.

Contributed by Alex Najibi

BACKGROUND: Although immune checkpoint inhibitors (ICIs) have long half-lives, preclinical and retrospective clinical studies across multiple tumor types suggest that the time-of-day of ICI infusion may influence therapeutic efficacy by aligning initial drug exposure with circadian peaks in T-cell responsiveness. The immunological basis of this phenomenon and its clinical relevance in hepatocellular carcinoma (HCC) remains unknown. METHODS: We followed patients with advanced HCC receiving ICI therapy at Johns Hopkins from 2021 to 2025, classifying them into a morning (first treatment before 12:00 hours) or afternoon (first treatment after 12:00 hours) group. We assessed clinical outcomes and compared immunological responses from baseline to early-on-treatment by profiling peripheral blood mononuclear cells using cytometry by time-of-flight and plasma cytokines using a 39-plex Luminex assay. RESULTS: Our cohort included 84 patients, 39 of whom received their first infusion in the morning. There were no statistically significant differences in baseline demographic or clinical characteristics between patients initiating therapy in the morning versus afternoon. The morning group had superior progression-free survival (multivariable HR 0.50, 95% CI 0.30 to 0.84, p<0.01) and higher odds of treatment response (multivariable OR 3.26, 95% CI 1.08 to 10.90, p<0.05), with no significant increase in immune-related adverse events. The timing of subsequent infusions after the first dose had no impact on outcomes. Immunological responses diverged after the initial dose, with morning-treated patients showing reduced interleukin (IL)-6 levels (p<0.01) and greater expansion of cytotoxic central memory CD8+ T_cells (p=0.01) as well as cytotoxic effector and effector memory CD8+ T_cells (p=0.06). CONCLUSIONS: Morning first-dose infusion of ICIs in HCC was associated with improved clinical outcomes and distinct immune responses, including reduced IL-6 signaling and expansion of cytotoxic central memory CD8+ T cells. These findings suggest that the timing of the initial infusion can imprint an immunological program that shapes subsequent antitumor immunity, providing a mechanistic rationale for strategically scheduling ICI administration.

Author Info: (1) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. (2) Sidney

Author Info: (1) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. (2) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (3) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (4) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (5) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (6) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (7) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (8) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. (9) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (10) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (11) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (12) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (13) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (14) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (15) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (16) F Hoffmann-La Roche Ltd, Basel, Switzerland. (17) F Hoffmann-La Roche Ltd, Basel, Switzerland. Genentech Inc, South San Francisco, California, USA. (18) Genentech Inc, South San Francisco, California, USA. (19) Genentech Inc, South San Francisco, California, USA. (20) Genentech Inc, South San Francisco, California, USA. (21) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. Ludwig Institute for Cancer Research, Baltimore, Maryland, USA. (22) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (23) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (24) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (25) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (26) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (27) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA. (28) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA mark.yarchoan@jhmi.edu mnakaza2@jhmi.edu. (29) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA mark.yarchoan@jhmi.edu mnakaza2@jhmi.edu.

Identification of cycling regulatory T cell precursors as conductors of immune escape during breast carcinoma progression Spotlight 

Using single-cell and spatial transcriptomics in human and rat models, Bui et al. mapped immune remodeling of normal breast, pre-malignant (DCIS) , and invasive (IBC) breast cancer and identified a proliferative FOXP3int MKI67hi cycling Treg (cycTreg) subset. CycTregs emerged at the DCIS-IBC junction, expanded in IBC, and predicted CD8+ infiltration, TCR diversity, disease-specific survival, and DCIS recurrence. CycTreg abundance correlated with CLEC10A+ cDC2s, high HLA class II, and IL-33-producing CAFs. OX40 agonism plus anti-PD-L1 or IL-33 blockade reduced cycTreg, remodeled CAF states, and restored antitumor immunosurveillance.

Contributed by Shishir Pant

Using single-cell and spatial transcriptomics in human and rat models, Bui et al. mapped immune remodeling of normal breast, pre-malignant (DCIS) , and invasive (IBC) breast cancer and identified a proliferative FOXP3int MKI67hi cycling Treg (cycTreg) subset. CycTregs emerged at the DCIS-IBC junction, expanded in IBC, and predicted CD8+ infiltration, TCR diversity, disease-specific survival, and DCIS recurrence. CycTreg abundance correlated with CLEC10A+ cDC2s, high HLA class II, and IL-33-producing CAFs. OX40 agonism plus anti-PD-L1 or IL-33 blockade reduced cycTreg, remodeled CAF states, and restored antitumor immunosurveillance.

Contributed by Shishir Pant

ABSTRACT: Immune escape during the ductal carcinoma in situ (DCIS)-to-invasive breast cancer (IBC) transition shapes tumor evolution. Through transcriptomic mapping of the immune landscapes of normal breast, DCIS, and IBC from large patient cohorts, we identified T and myeloid cells as the primary distinguishing features between DCIS and IBC. We discovered cycling regulatory T cells (cycTreg) as an orchestrator of immunosuppression in IBC. cycTreg frequency predicts cytotoxic CD8(+), TCR diversity, disease-specific survival in IBC, and recurrence in DCIS. In a rat model of breast cancer, we demonstrated that cycTreg act as precursors to mature Treg and are inducible by tumor-localized type 2 dendritic cells. Profiling of tumors subjected to OX40 and PD-L1 therapies revealed an IL-33-mediated fibroblast-cycTreg signaling loop, the disruption of which enhances intratumoral antigen-experienced CD8(+) effectors and systemic immunosurveillance. Our study defines cycTreg as critical inducers of immune escape and promising immuno-oncology targets in breast cancer.

Author Info: (1) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of

Author Info: (1) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (2) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (3) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (4) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (5) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (6) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (7) Department of Data Science, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Biostatistics, Harvard T. H. Chan School of Public Health, Boston, MA 02115, USA. (8) Department of Data Science, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Biostatistics, Harvard T. H. Chan School of Public Health, Boston, MA 02115, USA. (9) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (10) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (11) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (12) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. (13) Duke Cancer Institute, Duke University School of Medicine, Durham, NC 27705, USA. (14) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA. (15) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA. (16) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA. (17) Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA. (18) Sutter Institute for Medical Research, Roseville, CA 95661, USA. (19) Sutter Institute for Medical Research, Roseville, CA 95661, USA. (20) Sutter Institute for Medical Research, Roseville, CA 95661, USA. (21) Sutter Institute for Medical Research, Roseville, CA 95661, USA. (22) Institute for Precision Medicine, University of Pittsburgh, Pittsburgh, PA 15260, USA. (23) Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. (24) Department of Surgery, Duke University School of Medicine, Durham, NC 27708, USA. (25) Department of Surgery, Washington University School of Medicine, St. Louis, MO 63108, USA. (26) Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggi, Republic of Korea. (27) Institute for Precision Medicine, University of Pittsburgh, Pittsburgh, PA 15260, USA; UPMC Hillman Cancer Center, Pittsburgh, PA 15213, USA. (28) Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. (29) UPMC Hillman Cancer Center, Pittsburgh, PA 15213, USA. (30) Sutter Institute for Medical Research, Roseville, CA 95661, USA. (31) Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. (32) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA. (33) Department of Surgery, Duke University School of Medicine, Durham, NC 27708, USA. (34) Department of Data Science, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Biostatistics, Harvard T. H. Chan School of Public Health, Boston, MA 02115, USA. (35) Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA. Electronic address: kornelia_polyak@dfci.harvard.edu.

Immune-induced TCR-like antibodies regulate specific T cell response in mice Spotlight 

Kishida et al. showed that immune-induced TCR-like antibodies (iTabs) – antibodies that are specific to an antigen peptide–MHC-II complex – were produced during helper T cell responses to immunization with various antigens. These iTabs induced antigen-dependent depletion of target cells, blocked TCR recognition of specific peptide–MHC-II complexes, and prevented activation of antigen-specific T cells, but only when the presented peptides contained specific flanking residues. In a mouse model, treatment with iTabs or immunization with a peptide that induced iTabs effectively limited the development of autoimmune encephalomyelitis.

Contributed by Lauren Hitchings

Kishida et al. showed that immune-induced TCR-like antibodies (iTabs) – antibodies that are specific to an antigen peptide–MHC-II complex – were produced during helper T cell responses to immunization with various antigens. These iTabs induced antigen-dependent depletion of target cells, blocked TCR recognition of specific peptide–MHC-II complexes, and prevented activation of antigen-specific T cells, but only when the presented peptides contained specific flanking residues. In a mouse model, treatment with iTabs or immunization with a peptide that induced iTabs effectively limited the development of autoimmune encephalomyelitis.

Contributed by Lauren Hitchings

ABSTRACT: Antigen-specific regulation of T cell response is crucial for limiting hyperimmune response. However, the molecular mechanisms governing specific immune regulation remain unclear. In this study, we discover that antibodies specific to the antigen peptide-MHC class II complex are produced during helper T cell responses to various antigens, including hen egg lysozyme and proteolipid protein peptide. These antibodies specifically inhibit T cell receptor (TCR) recognition of MHC class II molecules presenting specific antigen peptide. We term these antibodies 'immune-induced TCR-like antibodies' or iTabs. Immunization with peptides containing flanking residues induces iTabs whereas immunization with peptides lacking flanking residues does not. Furthermore, we show that immunization with iTab-inducible peptide or iTab treatment suppress autoimmune disease development in a mouse model of experimental autoimmune encephalomyelitis. Thus, our findings provide a strategy for suppressing antigen-specific helper T cell responses using specific peptides, potentially controlling autoimmune diseases.

Author Info: (1) Department of Immunochemistry, Research Institute for Microbial Diseases, The University of Osaka, Suita, Osaka, Japan. (2) Biostructural Mechanism Group, RIKEN SPring-8 Center

Author Info: (1) Department of Immunochemistry, Research Institute for Microbial Diseases, The University of Osaka, Suita, Osaka, Japan. (2) Biostructural Mechanism Group, RIKEN SPring-8 Center, Hyogo, Japan. (3) Department of Drug Target Protein Research, Shinshu University School of Medicine, Matsumoto, Nagano, Japan. Department of Structural Biology and Biochemistry, Institute of New Industry Incubation, Institute of Science Tokyo, Tokyo, Japan. (4) Department of Immunochemistry, Research Institute for Microbial Diseases, The University of Osaka, Suita, Osaka, Japan. Laboratory for Innate Immune Systems, Department of Microbiology and Immunology, Graduate School of Medicine, The University of Osaka, Suita, Osaka, Japan. (5) Biostructural Mechanism Group, RIKEN SPring-8 Center, Hyogo, Japan. Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, Japan. (6) Department of Drug Target Protein Research, Shinshu University School of Medicine, Matsumoto, Nagano, Japan. Department of Structural Biology and Biochemistry, Institute of New Industry Incubation, Institute of Science Tokyo, Tokyo, Japan. (7) Department of Immunochemistry, Research Institute for Microbial Diseases, The University of Osaka, Suita, Osaka, Japan. arase@biken.osaka-u.ac.jp. Laboratory of Immunochemistry, WPI Immunology Frontier Research Center, The University of Osaka, Suita, Osaka, Japan. arase@biken.osaka-u.ac.jp. Center for Advanced Modalities and DDS, The University of Osaka, Suita, Osaka, Japan. arase@biken.osaka-u.ac.jp. Center for Infectious Disease Education and Research, The University of Osaka, Suita, Osaka, Japan. arase@biken.osaka-u.ac.jp.

PD-1 antibody-bound progenitor-exhausted CD8+ T cells in lymph nodes boost PD-1-blockade anti-tumor immunity in gastrointestinal cancer

Spotlight 

Utilizing scRNA/TCRseq, CITEseq, and a novel assay for cell-bound anti-PD-1 to study the dynamics of T cells targeted by anti-PD-1, Nose and Yasumizu et al. first found that abundance of progenitor-exhausted CD8+ T cells (Tpex) in metastasis-free lymph nodes (LNs), but not tumors or metastatic LNs, correlated with better prognosis in patients with anti-PD-1-naive gastric cancer. Anti-PD-1 promoted the proliferation of anti-PD-1high-bound Tpex in LNs, and clonotypes overlapped with intratumoral anti-PD-1-bound exhausted T cells (Tex), suggesting that anti-PD-1high-bound Tpex migrate to the tumor, where they differentiate into Tex.

Contributed by Ute Burkhardt

Utilizing scRNA/TCRseq, CITEseq, and a novel assay for cell-bound anti-PD-1 to study the dynamics of T cells targeted by anti-PD-1, Nose and Yasumizu et al. first found that abundance of progenitor-exhausted CD8+ T cells (Tpex) in metastasis-free lymph nodes (LNs), but not tumors or metastatic LNs, correlated with better prognosis in patients with anti-PD-1-naive gastric cancer. Anti-PD-1 promoted the proliferation of anti-PD-1high-bound Tpex in LNs, and clonotypes overlapped with intratumoral anti-PD-1-bound exhausted T cells (Tex), suggesting that anti-PD-1high-bound Tpex migrate to the tumor, where they differentiate into Tex.

Contributed by Ute Burkhardt

ABSTRACT: While progenitor-exhausted T cells (Tpex) expressing TCF1 and PD-1 are crucial for the therapeutic effect of immune checkpoint inhibitors (ICIs) with therapeutic anti-PD-1 antibodies (aPD-1), the dynamics of ICI-bound Tpex are not fully understood. In this study, we investigate ICI-bound T cells in detail using combined sequencing analysis at the single-cell level. By analyzing samples from gastrointestinal cancer patients with or without ICI treatment, we find that Tpex are enriched in proximal lymph nodes (LNs) and proliferate at a high rate after ICI treatment. Importantly, aPD-1 high-bound Tpex in LNs share T-cell receptor clonotypes with intratumoral exhausted CD8(+) T cells (Tex), suggesting their migration to tumor sites after ICI treatment. This study thus provides new insights into how ICIs enhance anti-tumor immunity by acting on Tpex in LNs, deepening our understanding of the cellular mechanisms underlying ICI therapy.

Author Info: (1) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. Department of Clinical Research in Tumor Immunology, Graduate Sch

Author Info: (1) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. (2) Experimental Immunology, WPI Immunology Frontier Research Center, The University of Osaka, Suita, Japan. Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), The University of Osaka, Suita, Japan. (3) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. tsaito@gesurg.med.osaka-u.ac.jp. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. tsaito@gesurg.med.osaka-u.ac.jp. (4) Experimental Immunology, WPI Immunology Frontier Research Center, The University of Osaka, Suita, Japan. (5) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. (6) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. (7) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (8) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (9) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (10) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (11) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (12) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (13) Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. Pharmaceutical Research Division, Shionogi & Co., Ltd., Toyonaka, Japan. (14) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (15) Department of Pathology, Institute of Medical Science (Medical Research Center), Tokyo Medical University, Tokyo, Japan. (16) Experimental Immunology, WPI Immunology Frontier Research Center, The University of Osaka, Suita, Japan. Department of Basic Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Osaka, Japan. (17) Experimental Immunology, WPI Immunology Frontier Research Center, The University of Osaka, Suita, Japan. (18) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, The University of Osaka, Suita, Japan. (19) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan. (20) Department of Gastroenterological Surgery, Graduate School of Medicine, The University of Osaka, Suita, Japan.

Acute and chronic infections drive distinct trajectories in human memory CD4+ T cell formation

Spotlight 

Comparing CD4+ T cells generated during acute or chronic hepatitis C virus (HCV) infection, Reinscheid and Weisser et al. evaluated patient samples and found that acute infection generated various subsets of progenitor CD4+ T cells, including subsets also observed in chronic infection. In chronic infection, a subset of stem-like/resting Bcl-2+ CD4+ T cells likely gave rise to a subset of T-bet+ effector CD4+ T cells. In patients treated with DAA to clear the virus, the effector subset essentially disappeared, while the stem-like subset formed a functional long-term memory pool that was distinct from the memory pools that formed after spontaneous HCV clearance.

Contributed by Lauren Hitchings

Comparing CD4+ T cells generated during acute or chronic hepatitis C virus (HCV) infection, Reinscheid and Weisser et al. evaluated patient samples and found that acute infection generated various subsets of progenitor CD4+ T cells, including subsets also observed in chronic infection. In chronic infection, a subset of stem-like/resting Bcl-2+ CD4+ T cells likely gave rise to a subset of T-bet+ effector CD4+ T cells. In patients treated with DAA to clear the virus, the effector subset essentially disappeared, while the stem-like subset formed a functional long-term memory pool that was distinct from the memory pools that formed after spontaneous HCV clearance.

Contributed by Lauren Hitchings

ABSTRACT: Virus-specific CD4(+) T cells are essential for coordinating adaptive immunity during infection, but their differentiation and maintenance in chronic infection remain unclear. Using human hepatitis C virus (HCV) infection as a model, we assessed the determinants of virus-specific CD4(+) T cell immunity in acute, spontaneously cleared, chronic, and therapeutically cured infections. During acute infection, multiple subsets of progenitor CD4(+) T cells emerged, including subsets that are also found in chronic infection. In chronic infection, stem-like Bcl-2(+) CD4(+) T cells and T-bet(+) effector CD4(+) T cells existed in a progenitor/progeny relationship. Following therapy-mediated HCV cure, these cells retained their chronic signature but formed a stable memory pool that persisted for years and was distinct from HCV-specific CD4(+) T cell memory after spontaneous clearance. Collectively, our findings highlight differences in CD4(+) T cell fates that depend on infection outcomes and reveal common principles of CD4(+) and exhausted CD8(+) T cell maintenance during and after chronic infection.

Author Info: (1) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany

Author Info: (1) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany. (2) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany. (3) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. (4) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. (5) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. (6) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany. (7) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. (8) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany. (9) Institute for Transfusion Medicine and Gene Therapy, Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany. (10) Institute of Virology, Medical Faculty and University Hospital DŸsseldorf, Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (11) Institute of Virology, Medical Faculty and University Hospital DŸsseldorf, Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (12) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany. (13) Institute of Virology, Medical Faculty and University Hospital DŸsseldorf, Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (14) Institute of Molecular and Clinical Ophthalmology Basel, Basel, Switzerland. (15) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany; German Cancer Consortium (DKTK), Heidelberg, Germany, partner site Freiburg, Freiburg, Germany; Signaling Research Centers BIOSS and CIBSS, Freiburg, Germany. (16) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. (17) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. Electronic address: robert.thimme@uniklinik-freiburg.de. (18) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. Electronic address: tobias.boettler@uniklinik-freiburg.de. (19) Department of Medicine II, Medical Center - University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, Freiburg, Germany. Electronic address: maike.hofmann@uniklinik-freiburg.de.

Precancerous niche remodelling dictates nascent tumour persistence Spotlight 

Using the DEN carcinogenesis model, Skrupskelyte and Arias et al. showed that early tumor persistence depended on the formation of a fibrotic precancerous niche. Stress responses in nascent epithelial lesions activated EGFR signaling, induced SOX9⁺, recruited PDGFRαlow fibroblasts, and drove the formation of a fibronectin (FN1)-rich niche that promoted tumor growth. Tumor-derived stroma alone was sufficient to impose tumor traits in normal epithelium. Inhibition of either fibronectin fibrillogenesis or EGFR signaling prevented niche formation and reduced tumor burdens. Heterogeneous AREG+ (an EGF ligand) and/or SOX9+ populations were present in early human oesophageal carcinoma.

Contributed by Shishir Pant

Using the DEN carcinogenesis model, Skrupskelyte and Arias et al. showed that early tumor persistence depended on the formation of a fibrotic precancerous niche. Stress responses in nascent epithelial lesions activated EGFR signaling, induced SOX9⁺, recruited PDGFRαlow fibroblasts, and drove the formation of a fibronectin (FN1)-rich niche that promoted tumor growth. Tumor-derived stroma alone was sufficient to impose tumor traits in normal epithelium. Inhibition of either fibronectin fibrillogenesis or EGFR signaling prevented niche formation and reduced tumor burdens. Heterogeneous AREG+ (an EGF ligand) and/or SOX9+ populations were present in early human oesophageal carcinoma.

Contributed by Shishir Pant

ABSTRACT: Interactions between mutant cells and their environment have a key role in determining cancer susceptibility(1-3). However, understanding of how the precancerous microenvironment contributes to early tumorigenesis remains limited. Here we show that newly emerging tumours at their most incipient stages shape their microenvironment in a critical process that determines their survival. Analysis of nascent squamous tumours in the upper gastrointestinal tract of the mouse reveals that the stress response of early tumour cells instructs the underlying mesenchyme to form a supportive 'precancerous niche', which dictates the long-term outcome of epithelial lesions. Stimulated fibroblasts beneath emerging tumours activate a wound-healing response that triggers a marked remodelling of the underlying extracellular matrix, resulting in the formation of a fibronectin-rich stromal scaffold that promotes tumour growth. Functional heterotypic 3D culture assays and in vivo grafting experiments, combining carcinogen-free healthy epithelium and tumour-derived stroma, demonstrate that the precancerous niche alone is sufficient to confer tumour properties to normal epithelial cells. We propose a model in which both mutations and the stromal response to genetic stress together define the likelihood of early tumours to persist and progress towards more advanced disease stages.

Author Info: (1) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. gs463@cam.ac.uk. Department of Physiology, Development and Neuroscience,

Author Info: (1) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. gs463@cam.ac.uk. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. gs463@cam.ac.uk. (2) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. RhyGaze, Basel, Switzerland. (3) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. (4) Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, Dresden, Germany. Center for Systems Biology, Dresden, Germany. (5) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. (6) Gurdon Institute, University of Cambridge, Cambridge, UK. (7) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. (8) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. (9) Wellcome Sanger Institute, Hinxton, UK. Cambridge Institute of Science, Altos Labs, Cambridge, UK. (10) Wellcome Sanger Institute, Hinxton, UK. (11) Wellcome Sanger Institute, Hinxton, UK. (12) Wellcome Sanger Institute, Hinxton, UK. (13) University Hospital Carl Gustav Carus Dresden, Faculty of Medicine of TUD Dresden University of Technology, Dresden, Germany. Institute of Pathology, University Hospital CGC Dresden, TU Dresden, Dresden, Germany. (14) Institute of Anatomy, Faculty of Medicine of TUD, University of Technology, Dresden, Germany. (15) Institute of Anatomy, Faculty of Medicine of TUD, University of Technology, Dresden, Germany. (16) Department of Gastroenterology, Guy's and St. Thomas' Hospital, London, UK. (17) Wellcome Sanger Institute, Hinxton, UK. Addenbrooke's Hospital, Cambridge University Hospital NHS Trust, Cambridge, UK. (18) Wellcome Sanger Institute, Hinxton, UK. Department of Oncology, University of Cambridge, Hutchison Research Centre, Cambridge Biomedical Campus, Cambridge, UK. (19) Max Planck Institute for the Physics of Complex Systems, Dresden, Germany. Arnold Sommerfeld Center for Theoretical Physics, Ludwigs-Maximilians-UniversitŠt Munchen, Munich, Germany. (20) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. Gurdon Institute, University of Cambridge, Cambridge, UK. Department of Applied Mathematics and Theoretical Physics, Centre for Mathematical Science, University of Cambridge, Cambridge, UK. (21) Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK. mpa28@cam.ac.uk. Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK. mpa28@cam.ac.uk.

Nous-209 neoantigen vaccine for cancer prevention in Lynch syndrome carriers: a phase 1b/2 trial Spotlight 

D’Alise and Willis et al. presented results from a phase 1b/2 single-arm trial of 45 Lynch syndrome (LS) carriers treated with Nous-209 – an IM, heterologous, prime/boost, virus-based vaccine encoding 209 frameshift peptides (FSPs) shared across neoplasms with microsatellite instability (MSI). Potent/durable vaccine-induced FSP-specific IFNγ-producing CD4+ and cytotoxic CD8+ T cell responses were observed in all 37 evaluable participants. Peptide–HLA predictions helped identify >100 FSPs, which were immunogenic in vitro and detected in datasets of LS MSI-high colorectal pre-cancers/cancers. The vaccine was safe, and no participants had advanced adenomas or CRC at the end of the study.

Contributed by Paula Hochman

D’Alise and Willis et al. presented results from a phase 1b/2 single-arm trial of 45 Lynch syndrome (LS) carriers treated with Nous-209 – an IM, heterologous, prime/boost, virus-based vaccine encoding 209 frameshift peptides (FSPs) shared across neoplasms with microsatellite instability (MSI). Potent/durable vaccine-induced FSP-specific IFNγ-producing CD4+ and cytotoxic CD8+ T cell responses were observed in all 37 evaluable participants. Peptide–HLA predictions helped identify >100 FSPs, which were immunogenic in vitro and detected in datasets of LS MSI-high colorectal pre-cancers/cancers. The vaccine was safe, and no participants had advanced adenomas or CRC at the end of the study.

Contributed by Paula Hochman

ABSTRACT: Cancer interception is a preventative approach aiming to reduce cancer incidence by targeting precancers and early-stage cancers. Lynch syndrome (LS) is a prevalent hereditary cancer syndrome affecting ~1 in 300 individuals, with an overall lifetime cancer risk as high as 80%. LS is caused by germline mutations in the DNA mismatch repair genes, leading to microsatellite instability (MSI) and accumulation of shared mutations. When these occur in coding regions, they generate frameshift peptides (FSPs). Nous-209 is a neoantigen-directed immunotherapy based on a heterologous prime boost using great ape adenovirus and modified vaccinia virus Ankara encoding 209 FSPs shared across MSI neoplasms. We present the results from cohort 1 of a phase 1b/2 single-arm trial of Nous-209 for cancer interception in LS carriers (n = 45). Safety and immunogenicity were coprimary endpoints. Safety was assessed in 45 participants. Vaccination was safe with no intervention-related serious adverse events (AEs). The most common AEs were injection-site reactions (any grade in 91% of participants after prime and 76% after boost with no grade 3) and fatigue (any grade in 80% after prime and 53% after boost with 4% grade 3 after prime or after boost). Neoantigen-specific immune responses were observed after vaccination in 100% of evaluable participants (n = 37), with induction of potent T cell immunity (mean response at peak of ~1,100 interferon-γ spot-forming cells per million peripheral blood mononuclear cells). The immune response was durable and detectable at 1 year in 85% of participants. Both CD8+ and CD4+ T cells were induced, recognizing multiple FSPs. Peptide-human leukocyte antigen predictions allowed the identification of >100 immunogenic FSPs with demonstration of cytotoxic activity in vitro. Immunogenic FSPs were found in independent datasets of LS MSI colorectal precancers and cancers. These results highlight Nous-209 ability to efficiently stimulate immunity against neoantigens in LS, supporting its development for cancer interception (ClinicalTrials.gov identifier: NCT05078866 ).

Author Info: (1) Nouscom SRL, Rome, Italy. (2) Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (3) Department of Clinical C

Author Info: (1) Nouscom SRL, Rome, Italy. (2) Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (3) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (4) Department of Clinical Genetics, Fox Chase Cancer Center, Philadelphia, PA, USA. (5) University of Puerto Rico Medical Sciences Campus, San Juan, PR, USA. (6) City of Hope Comprehensive Cancer Center, Duarte, CA, USA. (7) Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (8) University of Puerto Rico Medical Sciences Campus, San Juan, PR, USA. (9) Nouscom SRL, Rome, Italy. (10) Nouscom SRL, Rome, Italy. (11) Nouscom SRL, Rome, Italy. (12) Nouscom SRL, Rome, Italy. (13) Nouscom SRL, Rome, Italy. (14) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (15) Nouscom SRL, Rome, Italy. (16) Nouscom AG, Basel, Switzerland. (17) Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (18) Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (19) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (20) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (21) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (22) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (23) Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, USA. (24) Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, USA. (25) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (26) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (27) Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, USA. Department of Surgery, Walter Reed National Military Medical Center, Bethesda, MD, USA. (28) Nouscom SRL, Rome, Italy. e.scarselli@nouscom.com. (29) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. EVilar@mdanderson.org.

Tissue-resident exhausted and memory CD8+ T cells have distinct ontogeny, function and role in disease

Spotlight 

Park et al. showed that chronic antigen exposure drove a distinct lineage of tissue-resident exhausted CD8+ T cells (TR-TEX) that was developmentally and functionally separate from tissue-resident memory (TRM) cells formed after antigen clearance. TR-TEX (Tox-dependent) and TRM (Tox-independent) cells shared residency features, but were governed by divergent transcriptional and epigenetic programs. TRM cells retained plasticity to differentiate into TEX cells under chronic stimulation, while committed TEX cells failed to generate TRM cells after antigen withdrawal. TR-TEX cells responded to PD-1 pathway blockade in vivo, and were associated with patient responses to ICB.

Contributed by Shishir Pant

Park et al. showed that chronic antigen exposure drove a distinct lineage of tissue-resident exhausted CD8+ T cells (TR-TEX) that was developmentally and functionally separate from tissue-resident memory (TRM) cells formed after antigen clearance. TR-TEX (Tox-dependent) and TRM (Tox-independent) cells shared residency features, but were governed by divergent transcriptional and epigenetic programs. TRM cells retained plasticity to differentiate into TEX cells under chronic stimulation, while committed TEX cells failed to generate TRM cells after antigen withdrawal. TR-TEX cells responded to PD-1 pathway blockade in vivo, and were associated with patient responses to ICB.

Contributed by Shishir Pant

ABSTRACT: The presence of CD8(+) T cells coexpressing residency and exhaustion molecules in chronic diseases often correlate with clinical outcomes; however, the relationship between these cells and conventional tissue-resident memory (T(RM)) cells or exhausted CD8(+) T (T(EX)) cells is unclear. Here we show that chronic antigen stimulation drives development of tissue-resident T(EX) (TR-T(EX)) cells that are distinct from T(RM) cells generated after antigen clearance. TR-T(EX) and T(RM) cells are regulated by different transcriptional networks with only TR-T(EX) cells being Tox-dependent for residency programming. While T(EX) cells (including TR-T(EX)) are unable to generate T(RM) cells after antigen withdrawal, T(RM) cells differentiate into T(EX) cells upon chronic antigen exposure. Cell-state-specific transcriptional signatures reveal a selective association of TR-T(EX) cells with patient responses to immune checkpoint blockade, and only TR-T(EX) but not T(RM) cells responded to PD-1 pathway inhibition in vivo. These data suggest that TR-T(EX) and T(RM) cells are developmentally divergent cell states that share a tissue-residency program but have distinct roles in disease control.

Author Info: (1) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. simone.park@pennmedicine.upen

Author Info: (1) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. simone.park@pennmedicine.upenn.edu. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. simone.park@pennmedicine.upenn.edu. (2) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (3) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (4) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (5) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (6) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Graduate Group in Biochemistry and Molecular Biophysics, University of Pennsylvania, Philadelphia, PA, USA. (7) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (8) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (9) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (10) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Medicine II (Gastroenterology, Hepatology, Endocrinology and Infectious Diseases), Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany. (11) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (12) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (13) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (14) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (15) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Dermatology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (16) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (17) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Division of Cardiovascular Medicine, Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, USA. (18) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Medicine, Massachusetts General Hospital, Boston, MA, USA. (19) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (20) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (21) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (22) Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Dermatology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (23) Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Dermatology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (24) Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Medicine: Hematology and Oncology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (25) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (26) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (27) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (28) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (29) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (30) Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (31) Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Department of Otorhinolaryngology: Head and Neck Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. (32) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. (33) Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. wherry@pennmedicine.upenn.edu. Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. wherry@pennmedicine.upenn.edu. Parker Institute for Cancer Immunotherapy at University of Pennsylvania, Philadelphia, PA, USA. wherry@pennmedicine.upenn.edu.

Predicability of PD-L1 expression in cancer cells based solely on H&E-stained sections

PD-L1 expression is an important biomarker for selecting patients who are eligible for immune checkpoint inhibitor (ICI) therapy. However, evaluating PD-L1 through immunohistochemistry often faces significant interobserver variability and requires considerable time and resources. Recent advancements in artificial intelligence (AI) have transformed the field of pathology, leading to more standardized and reproducible methods for biomarker quantification. In this study, we examine the application of AI-driven models, particularly deep learning algorithms, to predict PD-L1 expression directly from hematoxylin and eosin-stained histological slides. Several AI-based approaches have been studied, demonstrating high accuracy in estimating PD-L1 expression and predicting responses to ICIs across various cancer types. AI-driven assessments of PD-L1 have been shown to reduce the subjectivity associated with manual scoring methods, such as the Tumor Proportion Score and the Combined Positive Score. Moreover, integrating AI with multimodal data, including genomics, radiomics, and real-world clinical data, can further enhance predictive accuracy and improve patient stratification for immunotherapy. Finally, AI-driven computational pathology offers a transformative approach to biomarker evaluation, providing a faster, more objective, and cost-effective alternative to traditional methods, with significant implications for personalized oncology and precision medicine. Despite these promising results, several challenges remain to be addressed, such as the need for large-scale validation, standardization of AI models, and regulatory approvals for clinical implementation. Tackling these issues will be crucial for incorporating AI-based PD-L1 assessments into routine pathology workflows.

Author Info: (1) Department of Medical Sciences and Public Health, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. Department of Biology, College of Science and Technology, Temple Un

Author Info: (1) Department of Medical Sciences and Public Health, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA. (2) Department of Electrical and Electronic Engineering, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. (3) Medical Oncology Unit, University Hospital of Cagliari, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. (4) Medical Oncology Unit, University Hospital of Cagliari, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. (5) Department of Pathology, Ospedale Oncologico A. Businco, ARNAS G. Brotzu, Cagliari, Italy. (6) Department of Medical Sciences and Public Health, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. (7) Medical Oncology Unit, University Hospital of Cagliari, Universitˆ degli Studi di Cagliari, 09123 Cagliari, Italy. (8) National Cancer Institute, Kyiv, Ukraine. Associazione "Angela Serra" per la ricerca sul cancro, Modena, Italy. (9) Department of Medicine - DIMED; General Anatomic Pathology and Cytopathology Unit, Universitˆ degli Studi di Padova, 35121 Padova, Italy.

A microenvironment-driven HLA-II-associated insulin neoantigen elicits persistent memory T cell activation in diabetes Spotlight 

Using immunopeptidome analysis in T1D patients, Srivastava et al. discovered a cys-to-ser mutation at position 19 (C19S) of insulin, within an MHC-II binding region. C19S could be contextually elicited via oxidative stress in islets, representing a dominant transformation among single AA variants (SAVs) and post-translational modifications. The conversion was also potentiated in APCs by cytokine exposure. In NOD mice and humans, InsB12-20(C19S) elicited a unique corresponding T cell population that was present in the non-diabetic state, yet expanded with diabetes progression to a greater extent than native InsB12-20-specific T cells, and was enriched in activated, Th1-like memory cells.

Contributed by Morgan Janes

Using immunopeptidome analysis in T1D patients, Srivastava et al. discovered a cys-to-ser mutation at position 19 (C19S) of insulin, within an MHC-II binding region. C19S could be contextually elicited via oxidative stress in islets, representing a dominant transformation among single AA variants (SAVs) and post-translational modifications. The conversion was also potentiated in APCs by cytokine exposure. In NOD mice and humans, InsB12-20(C19S) elicited a unique corresponding T cell population that was present in the non-diabetic state, yet expanded with diabetes progression to a greater extent than native InsB12-20-specific T cells, and was enriched in activated, Th1-like memory cells.

Contributed by Morgan Janes

ABSTRACT: The antigenic landscape of autoimmune diabetes reflects a failure to preserve self-tolerance, yet how novel neoantigens emerge in humans remains incompletely understood. Here we designed an immunopeptidomics-based approach to probe HLA-II-bound, islet-derived neoepitopes in patients with type 1 diabetes. We uncovered a Cys_Ser transformation, conserved between mice and humans, that reshapes autoreactivity to insulin at the single-residue level. This transformation, which we call C19S, arises from oxidative remodeling of insulin in stressed pancreatic islets and also occurs in cytokine-activated antigen-presenting cells, contributing to a feed-forward loop of neoepitope formation and presentation. Despite involving just one amino acid, C19S is recognized by HLA-DQ8-restricted, register-specific CD4(+) T cells that expand at diabetes onset. These neoepitope-specific CD4(+) T cells lack regulatory potential but acquire a poised central memory phenotype that persists throughout disease progression. These findings reveal a distinct, microenvironment-driven route of neoantigen formation that fuels sustained autoreactivity in diabetes.

Author Info: (1) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Hum

Author Info: (1) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (2) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (3) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (4) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (5) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (6) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (7) Department of Internal Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA. (8) Department of Developmental Biology, Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, MO, USA. (9) Department of Immunology and Microbiology, Scripps Research Institute, La Jolla, CA, USA. (10) Department of Pediatrics, Division of Endocrinology, Diabetes, and Metabolism, Washington University School of Medicine, St. Louis, MO, USA. (11) Department of Pediatrics, Division of Endocrinology, Diabetes, and Metabolism, Washington University School of Medicine, St. Louis, MO, USA. (12) Department of Radiation Oncology, Weill Cornell Medicine, New York, NY, USA. (13) Department of Immunology and Microbiology, Scripps Research Institute, La Jolla, CA, USA. (14) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. (15) Department of Developmental Biology, Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, MO, USA. (16) Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University School of Medicine, St. Louis, MO, USA. (17) Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. Center for Systems Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. Cancer Immunology and Immunotherapy Program, UPMC Hillman Cancer Center, Pittsburgh, PA, USA. (18) Department of Neurology, Hope Center for Neurological Disorders, Knight Alzheimer's Disease Research Center, Washington University School of Medicine, St. Louis, MO, USA. (19) Department of Internal Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA. (20) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. (21) Department of Radiation Oncology, Weill Cornell Medicine, New York, NY, USA. Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY, USA. Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA. (22) Department of Immunology and Microbiology, Scripps Research Institute, La Jolla, CA, USA. (23) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. (24) Department of Pediatrics, Division of Endocrinology, Diabetes, and Metabolism, Washington University School of Medicine, St. Louis, MO, USA. (25) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. clichti@wustl.edu. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. clichti@wustl.edu. (26) Department of Pathology and Immunology, Division of Immunobiology, Washington University School of Medicine, St. Louis, MO, USA. wanx@wustl.edu. The Andrew M. and Jane M. Bursky Center for Human Immunology, Washington University School of Medicine, St. Louis, MO, USA. wanx@wustl.edu.

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