Cancer Immunology, Immunotherapy

BackgroundVaccination against SARS-CoV-2 is a highly effective strategy to protect against infection, which is predominantly mediated by vaccine-induced antibodies. Postvaccination antibodies are robustly produced by those with inflammatory bowel disease (IBD) even on immune-modifying therapies but are blunted by anti-TNF therapy. In contrast, T-cell response which primarily determines long-term efficacy against disease progression,, is less well understood. We aimed to assess the post-vaccination T-cell response and its relationship to antibody responses in patients with inflammatory bowel disease (IBD) on immune-modifying therapies. MethodsWe evaluated IBD patients who completed SARS-CoV-2 vaccination using samples collected at four time points (dose 1, dose 2, 2 weeks after dose 2, 8 weeks after dose 2). T-cell clonal analysis was performed by T-cell Receptor (TCR) immunosequencing. The breadth (number of unique sequences to a given protein) and depth (relative abundance of all the unique sequences to a given protein) of the T-cell clonal response were quantified using reference datasets and were compared to antibody responses. ResultsOverall, 303 subjects were included (55% female; 5% with prior COVID) (Table). 53% received BNT262b (Pfizer), 42% mRNA-1273 (Moderna) and 5% Ad26CoV2 (J&J). The Spike-specific clonal response peaked 2 weeks after completion of the vaccine regimen (3- and 5-fold for breadth and depth, respectively); no changes were seen for non-Spike clones, suggesting vaccine specificity. Reduced T-cell clonal depth was associated with chronologic age, male sex, and immunomodulator treatment. It was preserved by non-anti-TNF biologic therapies, and augmented clonal depth was associated with anti-TNF treatment. TCR depth and breadth were associated with vaccine type; after adjusting for age and gender, Ad26CoV2 (J&J) exhibited weaker metrics than mRNA-1273 (Moderna) (p=0.01 for each) or BNT262b (Pfizer) (p=0.056 for depth). Antibody and T-cell responses were only modestly correlated. While those with robust humoral responses also had robust TCR clonal expansion, a substantial fraction of patients with high antibody levels had only a minimal T-cell clonal response. ConclusionAge, sex and select immunotherapies are associated with the T-cell clonal response to SARS-CoV-2 vaccines, and T-cell responses are low in many patients despite high antibody levels. These factors, as well as differences seen by vaccine type may help guide reimmunization vaccine strategy in immune-impaired populations. Further study of the effects of anti-TNF therapy on vaccine responses are warranted.

BackgroundCancer immunotherapy often triggers immune-related adverse events (irAEs). Analysis of irAEs in large checkpoint inhibitor (CPI) trials has enhanced their management and demonstrated their prognostic value for treatment outcome. However, data on irAEs in non-standard cancer immunotherapies (CITs) are limited, and systematic exploration is lacking. Identifying predictive biomarkers for irAEs in these therapies is still emerging and essential for improving patient care. MethodsWe established a harmonized data mart from 27 early-phase CIT trials, encompassing 14 molecules with diverse mechanisms across various cancer indications. This dataset includes 3,608 patients, both CPI-naive and CPI-experienced, with detailed information on clinical data, tumor characteristics, soluble biomarkers, and genome-wide genotyping. We examined the occurrence of different irAEs and CIT molecules concerning incidence, severity, and onset. A meta-analysis was conducted to assess the association between risk factors and the time to onset of irAEs. Finally, we explored the predictive value of irAEs for clinical outcomes, specifically measured by progression-free survival (PFS). ResultsOur analysis reveals significant variation in irAE incidence and kinetics across CIT molecules. Common irAEs include hepatitis, rash, acute kidney injuries, and hypothyroidism, with hepatitis often severe and others mild. Hepatitis is frequently associated with immunocytokine treatment, while T-cell bispecifics (TCBs) are linked to organ-specific toxicities. Hepatic metastases correlate with hepatitis but inversely with rash; elevated liver enzymes are associated with hepatitis, and high ferritin levels with acute kidney injury risk. Higher myeloid cell counts are associated with reduced rash likelihood. No tumor microenvironment (TME) associations were found, and polygenic risk scores (PGS) show limited utility in our setting. Rash correlates with improved outcomes, whereas hepatitis is associated with a poorer prognosis, independent of baseline prognostic state assessed by the Real World Prognostic score (ROPRO). ConclusionsThese findings highlight the complexity of immune toxicities in early-phase trials, emphasizing the importance of the CIT class, as well as the roles of tumor burden, metastasis sites, and systemic immune state in the development of irAEs. Additionally, the observed association between skin toxicities and improved PFS suggests that skin toxicity could serve as a marker of systemic immune activation across immunotherapy contexts. Key messagesWhat is already known on this topic[bullet] Cancer immunotherapy can induce immune-related adverse events (irAEs); their management and prognostic significance have advanced thanks to data from large checkpoint inhibitor trials. What this study adds[bullet] This study reveals the complexity of irAEs in early-phase pan-immunotherapy trials, highlighting the impact of tumor burden, metastasis sites, and systemic immune state, while identifying skin toxicity as a potential surrogate marker for improved patient outcomes. How this study might affect research, practice or policy[bullet] Our study lays a foundation for pan-immunotherapy irAE research, offering insights for clinicians and drug developers to assess risk profiles and guide the design of future trials for new immunotherapies.

Extreme fatigue is a clinical symptom that affects >50% of individuals with Inflammatory Bowel Disease (IBD), with a similar prevalence across many common immune-mediated inflammatory diseases (IMIDs). Despite its ubiquity, human scientific studies have yet to explain the mechanistic basis of this pervasive and complex symptom. One fundamental reason for this, is our inability to account for the clinical heterogeneity of fatigue with its multifactorial nature. We present the conceptual machine learning (ML) framework to dissect the complex nature of fatigue using one of the largest prospectively captured, real-world patient-reported outcome (PROs) on wellbeing from three contemporaneous cohorts (2020-present), totalling 2,970 responses from 2,290 participants across the UK and internationally, including non-IBD controls with 100 lines of clinical metadata. We systematically defined the (1) threshold of fatigue as our primary outcome ([≥]10/14 fatigue days) to build our ML approach, (2) utilised routinely available clinical data that can be used at a population-level analysis, (3) employed seven different ML methods with external validation in 3 different cohorts in UK, Spain and Australia (n=252), (4) employed Shapley Additive Explanations (SHAP) analysis to break down the clinical heterogeneity to allow the examination of clinical predictive factors at an individual level; and finally (5), investigate whether there are distinct clusters of fatigue patients. We found that ML models performed comparably (AUC/C-index [~]0.7) on external validation with SHAP analysis showing interpretable, individualised fatigue drivers and five distinct fatigue phenotypes, including a subgroup of young males with significantly lower fatigue burden. Our data therefore provides the ML roadmap to predict and deconstruct fatigue in IBD and potentially also more widely in IMIDs, enabling patient-level dissection beyond symptom-based classification with the ability to integrate deep molecular data. This is a step towards future clinical-scientific AI models with the immediate clinical application to stratify patients to human experimental studies to better understand the dominant mechanisms that drive fatigue at an individual level.

BackgroundThe tumor microenvironment is a site of intense metabolic competition between cancer cells and immune cells. In melanoma, tumor-intrinsic glutamine metabolism and the anti-tumor interferon-gamma (IFN{gamma}) immune response are critical opposing factors. We hypothesized that the interplay between these two pathways, termed a metabolic conflict, could be captured by a transcriptional signature to stratify patients and predict clinical outcomes. MethodsWe developed transcriptional signatures for glutamine metabolism and IFN{gamma} response pathways and calculated per-sample enrichment scores using Gene Set Variation Analysis (GSVA). We analyzed a discovery cohort of 469 melanoma patients from The Cancer Genome Atlas (TCGA-SKCM) to assess the signatures prognostic value. We then tested its predictive value for anti-PD-1 therapy in an independent validation cohort of 49 melanoma patients from GEO (GSE91061). ResultsIn the TCGA cohort, the glutamine metabolism and IFN{gamma} response scores were negatively correlated (Pearson r = -0.412), supporting the concept of a metabolic-immune conflict. Stratifying patients into four metabolic-immune groups revealed a trend towards improved overall survival for the Favorable group (Low Glutamine/High IFN{gamma}), but this did not reach statistical significance (log-rank p = 0.081). Cox proportional hazards models confirmed the signature lacked independent prognostic value when adjusted for clinical covariates. In the immunotherapy-treated validation cohort, the signature failed to predict clinical response to anti-PD-1 therapy (Chi-squared p = 0.706). ConclusionA transcriptional signature based on the interplay between glutamine metabolism and IFN{gamma} signaling, while biologically plausible, does not function as an independent prognostic or predictive biomarker in melanoma. Our findings underscore the value of rigorously testing well-founded hypotheses and highlight the complexity of translating metabolic-immune concepts into robust clinical tools.

Following the breakthrough of immune check point inhibitors (ICIs), a new era of immuno-oncology agents has emerged and established immunotherapy as a part of cancer treatment. Despite the improving outcomes of ICIs, many patients with initial response are known to develop acquired resistance later. There is increasing interest in utilizing other stimulatory means, such as anti-pathogen immune responses to induce anti-tumor immune responses. The immunostimulatory effects of anti-pathogen-treated tumors in combinations with ICI are known to potentially amplify anti-tumor immunity resulting in increased tumor responses and improved outcomes. Anti-pathogen-treated tumors can become immune-infiltrated "hot" tumors and demonstrate higher treatment response rates and improved survival. Our research group has previously demonstrated that tumors can be converted from "cold" to "hot" by intratumoral injection of a commercially available seasonal influenza vaccine. In continuation with our work, in deciphering the role of anti-viral immunity in the context of tumor immunology, we studied the role of inactivated SARS-CoV-2 virus as anti-tumor agent. Here we report that intratumoral injections of inactivated SARS-CoV-2 convert the immunologically cold tumors to hot by generating anti-tumor-mediated CD8+ T cells. Our findings suggest that inactivated SARS-CoV-2 can be used as an immune modulator in immunotherapy for melanoma and triple-negative breast cancer.

The composition of the tumour immune microenvironment (TIME) influences tumour evolution and responsiveness to immunotherapy. While longitudinal changes in TIME have been well-characterized in adult cancers, its dynamics in childhood cancers remain poorly documented, limiting our ability to predict treatment responses and tailor immunotherapeutic strategies. This study aimed to evaluate the plasticity of TIME in paediatric solid tumours, investigate its longitudinal evolution, and identify time-dependent immune alterations. Transcriptomic data from longitudinal samples of 27 paediatric patients (<21 years old) with relapsed or refractory solid tumours were analysed, encompassing 70 timepoints: 16 diagnoses and 54 successive relapses. TIME plasticity was assessed using gene expression clustering and immune cell infiltration enumeration. Patient-adjusted longitudinal analyses were performed using generalised linear mixed models (glmmSeq), adjusted for age and sex. Temporal associations of immune changes were further explored using dynamic regression models. Thirteen patients exhibited significant changes in their TIME profile, indicating high TIME plasticity. Over time, the TIME shifted toward a tolerogenic and immunosuppressive state, characterised by decreased activity in immune pathways (e.g., T cell receptor signalling) and enrichment of tolerogenic (e.g., macrophage differentiation) and oncogenic pathways (e.g., IL6-JAK-STAT3). The core enrichment of upregulated pathways contained key immunosuppressive factors: immune checkpoints (CTLA-4), tumour-associated macrophage activators (CSF1/CSF1R), T-regulatory cell activators (TGFB1), and immunosuppressive genes (IL10RA). This study provides evidence that the TIME in paediatric solid tumours is plastic and remodels towards immune depletion and tolerogenicity. This evolution may underlie treatment resistance and disease progression, underscoring the need for TIME-informed therapeutic approaches in paediatric oncology. Significance StatementThis article demonstrates the plasticity of the tumour immune micro-environment (TIME) of paediatric solid tumours throughout disease evolution. Longitudinal transcriptomic analyses of 70 tumour samples from 27 patients showed a progressive remodelling towards tolerogenicity and immune depletion. Key immunosuppressive factors, including immune checkpoints and tumour-associated macrophages, were identified as potential contributors to immune escape. These findings support the relevance of longitudinal immune monitoring in paediatric oncology and may inform future strategies for immunotherapeutic interventions.

Abstract/SummaryImmune checkpoint blockade can produce long-lasting responses in patients with metastatic melanoma; notably, combined CTLA-4/PD-1 blockade has been associated with approximately 52% melanoma specific 10-year survival (1). Yet, nearly half of patients experience minimal clinical benefit, and intensified regimens come with substantial risk of severe immune-related toxicity. The precise determinants of immunotherapy response are incompletely defined, reflecting a complex interplay between tumor biology and host immunity. This is especially consequential for patients whose disease progresses on checkpoint blockade, for whom effective salvage options are limited. In a series of patients with NRAS-mutated melanoma refractory to checkpoint inhibitors, we found that intratumoral administration of talimogene laherparepvec (T-VEC) combined with MEK inhibitor binimetinib induced exceptional clinical responses by amplification of pre-existing T cell responses and induction of de novo tumor-reactive immunity.

Improved understanding of the factors that underlie immune checkpoint inhibitor (ICI) response and toxicity are needed as only half of patients with metastatic melanoma respond, and 10-40% experience immune-related adverse events (irAEs). Modifying the gut microbiome could positively affect response to ICIs and reduce toxicities. Here, we sought to determine if the pre-treatment gut microbiome predicts ICI response or toxicity in the setting of metastatic melanoma. Melanoma patients (n=88) over 18 years of age, planning to receive ICI therapy enrolled in a prospective observational cohort study at The Ohio State University Comprehensive Cancer Center Skin Cancer Clinic. Patients taking corticosteroids for indications other than adrenal physiologic replacement were excluded. Stools were collected at baseline, within 10 days of an irAE as determined by CTCAE v 5.0 criteria, and at 12 weeks. ICI response and progression-free survival (PFS) were evaluated q12 weeks using Response Evaluation Criteria in Solid Tumors (RECIST v1.1). Metagenomic whole-genome shotgun sequencing of the microbiome was classified using MetaPhlAn4/HUMAnN3 and differential abundance analyzed with ANCOM-BC2. Of the 88 patients enrolled, 41 had metastatic disease and complete data. There were 25 participants classified as responders, defined as having complete response or partial response according to RECIST criteria, or stable disease with 6-month PFS. Grade [&ge;] 1 irAEs were observed in 15/41 participants. The abundance of Intestinimonas butyriciproducens (q-value = 0.002) and Longicatena caecimuris (q-value = 0.003) were enriched in responders, Tenericutes (q-value= 0.001) and Lachnospira sp. NSJ 43 (q-value =0.002) in non-responders. Blautia luti, as well as several other Lachnospiraceae, were associated with response and no irAE (response q-value = 0.02, no irAE q-value = 0.02). The association of response to ICIs with several taxa in the family Lachnospiraceae, a prevalent microbial family in the gut, is consistent with prior research, which has found that this family may influence treatment outcomes through various mechanisms, such as immune regulation, metabolism, and pathogen exclusion. While no statistical relationship was observed between response and irAEs in this cohort, the microbes associated with both could serve as biomarkers. Future studies to assign causal roles for (specific microbes) in response and toxicity could identify mechanisms to improve patient outcomes.

BackgroundVaccines targeting melanoma antigens can elicit CD8+ T cell responses, but a growing body of work suggests CD4+ T cells also play a role in tumor control. Induction of CD4+ cells may also support B cells in producing tumor antigen-specific antibodies (Abs). We investigated Abs induced by vaccination with a cocktail of six class II MHC-restricted melanoma peptides (6MHP) and the effect of adjuvant type on Ab isotypes. We hypothesized that the vaccines would induce Abs that respond to different epitopes on individual peptides and that IgG subclass distribution varies with different vaccine adjuvants. MethodsSera from patients who received a 6MHP vaccine were evaluated with enzyme-linked immunosorbent assays to map epitopes for polyclonal Ab responses to synthetic melanoma peptides (n=8). IgG subclasses of Ab responses to 6MHP were assessed in patients who received one of 4 adjuvants (Incomplete Freunds Adjuvant (IFA) alone, IFA + polyICLC, IFA + systemic mCy, or IFA + polyICLC + systemic mCy) to characterize IgG subclass distribution (n=2-14). Comparisons were evaluated using a Mann-Whitney rank sum test. ResultsEpitope mapping revealed that at least 50% of patients exhibiting Ab responses to melanoma peptides had responses to two or more epitopes on the same peptide, suggesting polyclonal antibody responses. Serum evaluation for IgG isotypes showed predominant induction of IgG1 and IgG3. Mean total IgG was highest when IFA and polyICLC were used in combination. Patients who received TLR3 agonist polyICLC had significantly higher concentrations of total IgG, IgG1, and IgG3 compared to patients who did not receive polyICLC. ConclusionsThese findings suggest that vaccine-induced Abs may respond to multiple epitopes within the same peptide, which may support creation of large antigen-Ab complexes, with promise to facilitate antigen uptake and presentation. Abs were predominantly IgG1 and IgG3, which are optimal for binding complement and supporting Ab-dependent cellular cytotoxicity. The findings also show that adding polyICLC to IFA can significantly enhance Ab responses. Collectively, this work underscores the immunologic potential of peptide-induced Abs and the importance of adjuvant selection in cancer vaccine design.

BACKGROUND AND AIMSVaccine-mediated immune responses in patients with inflammatory bowel disease (IBD) may be influenced by IBD therapies. We investigated in-depth humoral and T-cell responses to SARS-CoV-2 vaccination in IBD patients following three COVID-19 vaccine doses. METHODSImmune responses of 100 SARS-CoV-2-uninfected IBD patients on varying treatments were compared to healthy controls (n=35). Anti-S1/2 and anti-RBD SARS-CoV-2-specific antibodies, CD4+ and CD8+ T-cell responses were measured at baseline and at five time-points after COVID-19 vaccination. RESULTSAnti-S1/2 and anti-RBD antibody concentrations at [~]1 month after second dose vaccination were significantly lower in anti-TNF-treated patients compared to non-TNF IBD patients and healthy controls (126.4 vs 262.1 and 295.5, p<0.0001). Anti-S1/2 antibodies remained reduced in anti-TNF treated patients before and after the third dose (285.7 vs 365.3, p=0.03), although anti-RBD antibodies reached comparable titres to non-TNF patients. Anti-RBD antibodies were higher in the vedolizumab group than controls after second dose (4.2 vs 3.6, p=0.003). Anti-TNF monotherapy was associated with increased CD4+ and CD8+ T-cell activation compared to combination anti-TNF patients after second dose, but comparable after third dose. Overall, IBD patients demonstrated similar CD4+/CD8+ T-cell responses compared to healthy controls regardless of treatment regimen. CONCLUSIONSAnti-TNFs impaired antibody concentrations when compared to non-TNF patients and controls after two vaccine doses. These differences were not observed after the third vaccine dose. However, vaccine induced SARS-CoV-2-specific T cell responses are robust in anti-TNF-treated patients. Our study supports the need for timely booster vaccination particularly in anti-TNF treated patients to minimise the risk of severe SARS-CoV-2 infection.

Non-muscle invasive bladder cancer (NMIBC) constitutes a significant clinical challenge, with over 50% of patients experiencing poor clinical outcomes in the form of early recurrence or progression following treatment with Bacillus Calmette-Guerin (BCG) immunotherapy. The pre-treatment tumor immune microenvironment (TIME) is an established determinant of response to BCG. This study explores the spatial profiles of CD79a+ B cells, CD163+ M2-like macrophages, proliferating and tissue-resident phenotypes of T cells, along with PD-1/PD-L1 checkpoint expression in pre-BCG treatment tumors of 173 patients (139 males, 34 females). Multiplex immunofluorescence staining of a tumor tissue microarray, revealed elevated infiltration of CD79a+ B cells, CD163+ M2-like macrophages, CD103+ cells, and CD8+ T cells at the tumor invasive margins. Increased epithelial PD-L1 immune-checkpoint expression in tumors was observed in female and male patients who exhibited significantly shorter recurrence-free survival (RFS). Importantly, high CD79a+ B cell density in BCG-treated females in both stromal and epithelial compartments exhibited significantly shorter RFS and progression-free survival compared to males. Stromal CD79a+ B cell density was positively correlated with M2-like macrophages, CD8+ T cells, CD103+ cells and PD-1 expressing cells. CD79a+ B cells, CD103+ cells, and M2-like macrophage density were associated with higher grade and enriched in basal subtype tumor. This study highlights the significance of an understudied role of B cells and their cellular neighborhoods in the pre-treatment TIME and BCG-therapy response. Overall, findings from this study underscore the importance of considering sex-related immunobiological differences in the stromal compartments of bladder tumors towards the development of optimal therapeutic targeting strategies.

IntroductionLynch syndrome (LS) is the most common hereditary cause of colorectal cancer (CRC), increasing lifetime risk of CRC by up to 70%. Despite this higher lifetime risk, disease penetrance in LS patients is highly variable and most LS patients undergoing CRC surveillance will not develop CRC. Therefore, biomarkers that can correctly and consistently predict CRC risk in LS patients are needed to both optimize LS patient surveillance and help identify better prevention strategies that reduce risk of CRC development in the subset of high-risk LS patients. MethodsNormal-appearing colorectal tissue biopsies were obtained during repeat surveillance colonoscopies of LS patients with and without a history of CRC, healthy controls (HC), and patients with a history of sporadic CRC. Biopsies were cultured in an ex-vivo explant system and their supernatants were assayed via multiplexed ELISA to profile the local immune signaling microenvironment. High quality cytokine signatures were identified using rxCOV fidelity metric. These signatures were used to perform biomarker selection by computing their selection probability based on penalized logistic regression. ResultsOur study demonstrated that cytokine based local immune microenvironment profiling was reproducible over repeat visits and sensitive to patient LS-status and CRC history. Furthermore, we identified sets of biomarkers whose differential expression was predictive of LS-status in patients when compared to sporadic CRC patients and in identifying those LS patients with or without a history of CRC. Enrichment analysis based on these biomarkers revealed an LS and CRC status dependent constitutive inflammatory state of the normal appearing colonic mucosa. DiscussionThis prospective pilot study demonstrated that immune profiling of normal appearing colonic mucosa discriminates LS patients with a prior history of CRC from those without it, as well as patients with a history of sporadic CRC from HC. Importantly, it suggests existence of immune signatures specific to LS-status and CRC history. We anticipate that our findings have the potential to assess CRC risk in individuals with LS and help in preemptively mitigating it by optimizing surveillance and identifying candidate prevention targets. Further studies are required to validate our findings in an independent cohort of LS patients over multiple visits.

The development of cancer immunotherapies is hindered by the lack of human-relevant models that accurately translate to patient outcomes. We combine patient-derived colorectal tumor organoids (PDOs) and cancer-associated fibroblasts (CAFs) into floating extracellular matrix drops to form miniature colorectal tumors. These SHaking Organoid COcultures (SHOCOs) maintain immune cells in numbers, states and functional interactions that are more physiologically accurate than traditional PDO-based co-culture models. Immunocompetent SHOCOs treated with T-cell bispecific antibodies exhibited a robust anti-tumor response, in a concentration- and duration of treatment-dependent manner. By varying the stromal content, we found that fibroblasts present a physical barrier that hinders intratumoral T-cell infiltration. We also demonstrate that tumor-associated stroma can be exploited therapeutically in potentiating anti-tumor immune responses. SHOCOs could aid the battle against cancer both by providing fundamental insights into immune and stromal tumor biology, and by catalyzing the discovery of novel therapeutic approaches. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=143 SRC="FIGDIR/small/729459v1_ufig1.gif" ALT="Figure 1"> View larger version (78K): org.highwire.dtl.DTLVardef@1aed309org.highwire.dtl.DTLVardef@a11696org.highwire.dtl.DTLVardef@1d1d74dorg.highwire.dtl.DTLVardef@18e2c3f_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 0C_FLOATNO / Graphical Abstract: Illustration of the SHOCO Model. (A) Overview of the components used in creating SHOCOs and the complex tumor microenvironment they replicate. Epithelial cells (pink), CAFs (red), CD8 T cell (blue), CD4 T cell (green). Characteristics of SHOCOs: (B) Emphasizing the role of CAFs in contracting and creating a denser microenvironment. (C) CAFs act as a physical barrier, preventing CD8 T cells from reaching tumor niches. (D) SHOCOs maintain a substantial myeloid population (violet) capable of performing antigen presentation. (E) T cells within SHOCOs mount an anti-tumor cytotoxic immune response as a result of immunotherapy. Granzymes (orange), cleaved Caspase-3/7 (yellow). C_FIG

Chimeric antigen receptors (CARs) are artificial receptors introduced mainly into T cells. CAR-induced immune cell (CARi) products have achieved impressive success rates in treating some difficult-to-treat hematological malignancies. Here, we describe effects of the global COVID-19 pandemic on CARi publication landscape. Due to the pandemic, the total number of publications decreased in 2020 compared to 2019 in all fields of cancer immunotherapy except CARi. Nearly exponential increases in the number of CARi publications slowed-down in 2020 for the first time in the past 11 years. There were more CARi than coronavirus publications until 2020 when coronavirus publications increased over 5,000% compared to 2019 (575 publications in 2019 vs. 30,390 in 2020). Unlike cancer immunotherapy where the majority of the publications consist of conference abstracts and review articles, majority of the coronavirus publications are original research articles. There are more coronavirus publications in Pubmed than Embase. The opposite is true for CARi publications. Our analysis of the data from the FDA Adverse Event Reporting System (FAERS) show significantly higher death rate in patients treated with Kymirah than Yescarta (28.14% vs. 16.02%). Kymirah and Yescarta are the two main CAR T cell products for treatment of DLBCL and/or B-ALL. However, despite being highly significant, this result is not easily interpretable due to multiple confounding variables in the FAERS data. Our analysis additionally suggest that the significant effects of co-stimulatory domains (4-1BB vs. CD28) consistently reported in preclinical studies do not translate into clinical results. Our manual curation of the CARi publications in PubMed shows that only 5.2% of the publications report results from CARi clinical trials, although we found 663 clinical trials listed on ClinicalTrials.gov database. In conclusion, publication landscape in CARi as well as other fields of cancer immunotherapy has changed due to the global COVID-19 pandemic. This trend will likely continue in the near future. CARi research is now in need of increased measures by publishers to reduce repetitive and/or duplicate publications and more stringent criteria for data entry into public databases including PubMed, Embase, ClinicalTrials.gov, and FAERS to advance this important field of medical research.

ObjectiveLittle is known about the interplay between gut microbiome and SARS-CoV-2 vaccine immunogenicity. In this prospective observational study, we investigated associations between the gut microbiome, habitual dietary fibre intake, and mRNA vaccine-elicited immune responses, including anti-Spike IgG, avidity, and ACE-2 competition (surrogate neutralization). Design16S rRNA sequencing and short-chain fatty acid analyses were undertaken using stool samples collected from 48 healthy individuals at baseline and twelve-weeks after 1st BNT162b2 SARS-CoV-2 vaccine dose. Associations between gut microbiome data and SARS-CoV-2 spike and RBD IgG levels, competitive binding antibodies, and anti-SARS-CoV-2 spike total relative fractional avidity assays were evaluated. A validated dietary fibre intake food frequency questionnaire was also used to correlate habitual dietary fibre intakes with vaccine responses. ResultsOur data revealed several baseline bacterial taxa, including Prevotella, Haemophilus and Veillonella (p<0.01), associated with BNT162b2 vaccine responses. Several Bacteroides spp. (p<0.01) as well as Bifidobacterium animalis, (p=0.003), amongst others, were positively associated with antibody avidity. Conversely, concentrations of isovaleric and isobutyric acid were higher in individuals with the lowest SARS-CoV-2 vaccine responses (p<0.01). Classifying participants based on habitual dietary fibre intake identified distinct avidity responses. ConclusionWe showed associations between baseline gut microbiota composition and immunogenicity of BNT162b2 vaccine responses, particularly avidity maturation. We also demonstrate that branched-chain fatty acids and habitual dietary fibre intakes are associated with BNT162b2 vaccine immunogenicity. Together these findings indicate a link between gut microbiome, diet and antibody immunity to SARS-CoV-2 spike protein, suggesting interventions which modulate the gut microbiome could enhance COVID-19 vaccine responses. SIGNIFICANCE OF THIS STUDYO_ST_ABSWhat is already known on this subject?C_ST_ABSO_LIStrength and persistence of the SARS-CoV-2 BNT162b2 vaccine is variable between individuals. C_LIO_LITo date, only one study has demonstrated that baseline gut microbiota can predict SARS-CoV-2 vaccine response. C_LI What are the new findings?O_LIFor the first time we showed that the higher concentrations of branched-chain fatty acids, isovaleric and isobutyric acids, are negatively associated with SARS-CoV-2 BNT162b2 vaccine responses. C_LIO_LIWe revealed that habitual dietary fibre intake led to variability in the strength of antibody binding after the BNT162b2 vaccine. Specifically, high dietary fibre consumers displayed a significant increase in antibody avidity between in their 1st and 2nd dose. C_LI How this study might affect research, practice, or policyO_LIOur data suggests that therapeutic interventions which target the gut microbiome, including dietary modification, as well as pre-, pro-, and post-biotics, could enhance BNT162b2 vaccine immunogenicity, thus helping in the fight against COVID-19. C_LI

Melanoma remains one of the most treatment-refractory malignancies due to immune evasion, high mutational burden, and profound tumor heterogeneity. Although immune checkpoint inhibitors have transformed frontline management, a substantial proportion of patients develop resistance or experience relapse, underscoring the need for alternative and complementary immunotherapeutic strategies. Tumor-infiltrating lymphocyte (TIL) therapy and engineered viral vector-based immunotherapies represent mechanistically distinct yet clinically promising approaches for advanced melanoma. This systematic review and Bayesian meta-analysis evaluated the comparative efficacy of TIL therapy and engineered viral vector immunotherapies in advanced melanoma. A structured search of PubMed, Embase, Scopus, and Web of Science (2015-2025) identified 13 eligible studies, including four randomized controlled trials and nine prospective single-arm studies, reporting objective response rate (ORR), progression-free survival (PFS), overall survival (OS), and treatment-related adverse events. Eight studies met criteria for inclusion in the Bayesian quantitative synthesis of ORR outcomes. Risk of bias and certainty of evidence were assessed using Cochrane and GRADE frameworks. TIL therapy demonstrated substantial standalone efficacy, particularly in PD-1-refractory populations, with reported ORRs reaching 49%, median PFS of 7.2 months, and OS extending to 25.8 months. Viral vector-based therapies, including talimogene laherparepvec (T-VEC) and RP1, showed more modest monotherapy activity but demonstrated improved responses when combined with immune checkpoint inhibitors. Among the studies included in the Bayesian quantitative synthesis, the pooled ORR estimate was 37.8% (95% highest density interval [HDI]: 30.6%-45.3%). Sensitivity analysis excluding the small-sample Cui et al. (2022) study yielded a similar pooled estimate of 38.3% (95% HDI: 30.4%-46.2%). Exploratory meta-regression supported the overall robustness of the findings. Certainty of evidence for ORR was moderate, whereas survival and safety outcomes were downgraded due to heterogeneity, sparse reporting, and inconsistent endpoint definitions. Collectively, these findings support complementary rather than competing roles for TIL and engineered viral vector immunotherapies within evolving melanoma treatment paradigms. The results further highlight the potential importance of biomarker-guided sequencing strategies, including viral immune priming followed by adoptive cellular therapy, as a framework for optimizing personalized immunotherapy in both refractory and earlier-line melanoma settings.

Immune checkpoint blockade (ICB) therapies, targeting PD-1 or PD-L1, have transformed cancer treatment, particularly for aggressive cancers. However, many patients fail to benefit from ICBs due to tumor characteristics, including a non-inflammatory tumor microenvironment (TME) that impedes immune cell infiltration. This study investigated the potential of targeting the Atypical Chemokine Receptor 2 (ACKR2), known for scavenging CXCR3-related chemokines crucial for lymphocyte recruitment to tumors. Genetic targeting of ACKR2 in melanoma cells increased the release of essential chemokines associated with the inflamed TME. In mouse models, ACKR2 inhibition suppressed tumor growth, improved survival, and enhanced activated immune cell infiltration into the TME. Moreover, ACKR2 targeting synergized with anti-PD-1 therapy, overcoming resistance to anti-PD-1 and improving its efficacy. Analysis of melanoma patient data from The Cancer Genome Atlas (TCGA) revealed that patients with high levels of chemokines scavenged by ACKR2 had significantly better survival rates, with increased expression of NK cell and CD8 T cell markers indicating their presence in the TME. Notably, even in patients with high CD8 expression, those expressing low ACKR2 survived better than those expressing high ACKR2. This study emphasizes the clinical importance of targeting ACKR2 as an attractive strategy for the development of combination immunotherapies to treat cold tumors, which are clinically stratified to not be eligible for ICB-based therapy.

ObjectiveWe investigated UK wide inactivated influenza vaccine (IIV) uptake in adults with inflammatory bowel disease (IBD), the association between vaccination against influenza and IBD flare, and the effectiveness of IIV in preventing morbidity and mortality. DesignData for adults with IBD prior to 1st September 2018 were extracted from the Clinical Practice Research Datalink (CPRD) Gold, a database of electronic health records originated during routine care of patients in the UK. It is linked to hospitalization and mortality records. We calculated the proportion of patients vaccinated against seasonal influenza in the 2018-2019 influenza cycle. To investigate vaccine effectiveness, we calculated propensity score (PS) for vaccination and undertook Cox proportional hazard regression with inverse-probability treatment weighting on PS. We employed self-controlled case series (SCCS) to investigate the association between vaccination and IBD flare. ResultsData for 13,631 IBD patients (50.4% male, mean age 52.9 years) were included. Fifty percent were vaccinated during the influenza cycle while 32.1% were vaccinated before influenza virus circulated in the community. Vaccination was associated with a non-significant reduction in hospitalisation for pneumonia (aHR (95%CI) 0.52 (0.20-1.37), including in the influenza active period (aHR (95%CI) 0.48 (0.18-1.27)). Administration of the influenza vaccine was not associated with IBD flare. ConclusionThe uptake of influenza vaccine is low in IBD patients and the majority are not vaccinated before influenza virus circulates in the community. Vaccination with the IIV is not associated with IBD flare. These findings add to the evidence to promote vaccination in patients with IBD. Key messagesO_ST_ABSWhat is already known on the topicC_ST_ABSO_LIInactivated influenza vaccine is recommended in people with IBD treated with immune suppressing drugs. C_LIO_LIConcerns about influenza vaccine causing IBD flare and lack of data on the effectiveness of influenza vaccine in people with IBD are barriers to seasonal influenza vaccination in this population. C_LI What this study addsO_LIThe uptake of seasonal influenza vaccination is low in IBD patients. C_LIO_LISeasonal influenza vaccination is not associated with IBD flare and is more likely to prevent serious complications of influenza in this population. C_LI How this study might affect research, practice, or policyO_LIThis study provides new data on the uptake, effectiveness, and safety of influenza vaccine in people with IBD and adds to the accumulating evidence to promote vaccination in this population. C_LI

Therapeutic options for non-small cell lung cancer (NSCLC) treatment have changed dramatically in recent years with the advent of novel immunotherapeutic approaches. Among these, immune checkpoint blockade (ICB), using monoclonal antibodies, has shown tremendous promise in a small proportion of patients. In order to better predict patients that will respond to ICB treatment, biomarkers such as tumor-associated CD8+ T cell frequency, tumor checkpoint protein status and mutational burden have been utilized, however, with mixed success. In this study, we hypothesized that significantly altering the suppressive tumor immune landscape in NSCLC could potentially improve ICB efficacy. Using sub-therapeutic doses of our Salmonella typhimurium-based therapy targeting the suppressive molecule indoleamine 2,3-dioxygenase (shIDO-ST) in tumor-bearing mice, we observed dramatic changes in immune subset phenotypes that included increases in antigen presentation markers, decreased regulatory T cell frequency and overall reduced checkpoint protein expression. Combination shIDO-ST treatment with anti-PD-1/CTLA-4 antibodies enhanced tumor growth control, compared to either treatment alone, which was associated with a significant intratumoral influx of CD8+ and CD4+ T lymphocytes. These results suggest that the success of ICB therapy may be more accurately predicted by taking into account multiple factors such as potential for antigen presentation and frequency of suppressive immune subsets in addition to markers already being considered. Alternatively, combination treatment with agents such as shIDO-ST could be used to create a more conducive tumor microenvironment for improving response rates to immunotherapy.

Background The clinical significance of clonal hematopoiesis of indeterminate potential (CHIP) in melanoma remains incompletely defined, particularly with respect to CHIP genotype, clone size, and somatic mutations (e.g BRAF mutations). We integrated human cohort data and a syngeneic melanoma mouse model to evaluate whether CHIP is associated with melanoma risk, tumor growth, and differential clinical outcomes. Methods We analyzed CHIP prevalence and survival in a large treatment-unselected melanoma cohort (n=2,480), evaluated tumor growth in a syngeneic BRAF-mutant (BRAFmut) melanoma murine model of TET2-CHIP and DNMT3A-CHIP, and assessed survival outcomes in an immune checkpoint inhibitor (ICI)-treated advanced melanoma cohort (n=361). Associations with progression-free survival (PFS) and overall survival (OS) were evaluated using Kaplan-Meier analyses and multivariable Cox proportional hazards models. Results CHIP was enriched among patients with treatment-unselected melanoma compared with age/sex-matched healthy controls, and larger CHIP clone size showed an age-adjusted association with inferior OS. In a syngeneic BRAFmut melanoma murine model, TET2-CHIP, but not DNMT3A-CHIP, was associated with significantly increased primary melanoma tumor growth. Among patients with ICI-treated advanced melanoma, CHIP was associated with worse OS compared with patients without CHIP. TET2-CHIP had the strongest adverse association with survival, whereas DNMT3A-CHIP was not significantly associated with PFS or OS. Conclusions CHIP is enriched in melanoma and exploratory analyses demonstrate genotype-specific differences in melanoma tumor growth and clinical outcomes. These findings support further investigation of genotype-specific CHIP profiling as a potential biomarker for melanoma risk stratification and immunotherapy outcomes.