TUMOR BURDEN AND IMMUNE MICROENVIRONMENT INFLUENCE RESPONSE TO ANTI-BCMA BISPECIFIC ANTIBODIES IN RELAPSED/REFRACTORY MULTIPLE MYELOMA Elena Tofacchi Haematologica, 2026 Patients with triple-class refractory relapsed/refractory multiple myeloma (RRMM) face extremely poor outcomes, with median overall survival rarely exceeding 4–8 months despite intensive therapy. Against this dismal background, the advent of bispecific antibodies (BsAbs) has dramatically improved prognosis, producing deep and durable responses even in heavily pretreated individuals. Nevertheless, approximately 30–40% of patients exhibit primary resistance, underscoring the urgent need to identify predictive biomarkers of response to guide treatment selection and minimize unnecessary toxicity and cost.We conducted a retrospective study approved by the Ethics Committee of the University Hospital of Palermo to explore immunological and transcriptional determinants of response to new anti-MM BsAbs. In a cohort of 18 triple class exposed/refractory MM patients (7 treated with Teclistamab, 7 with Elranatamab, and 4 with Talquetamab), we integrated clinical, laboratory, and, where available single-cell RNA-seq/CITE-seq (SC) data to identify features associated with treatment resistance.Comparative analysis of 20 hematochemical parameters and 5 derived indices revealed that non responder patients presented an increased ISS stage (p=0.001), disease burden in term of monoclonal component (p=0.039), and a reduced Hemoglobin level (p:0.03). Interestingly, responders patients experienced a strong lymphocytes reduction between pre and post step-up dose (p<0.001). The latter in particular was associated with a significantly longest PFS (p=0.001, no events experienced after 18 months follow-up). To deeply understand changes in BM microenvironment, we performed a single-cell analysis of BMMCs from a subset of 10 patients at baseline, uncovering distinct immunological configurations between responders and non-responders. We analyzed a total of 39195 single cells were we found little to no deifferences in term of population abundances except with B cells (strongly reduced in non responders p<0.01). In term of transcriptome profile, we observed that responders displayed a more enhanced intrinsic anti-tumor immunity. In contrast, non-responders exhibited enrichment of exhausted T cells expressing HAVCR2 (TIM-3) and genes linked to oxidative stress and immune dysfunction, coupled with tolerogenic monocytes and upregulation of immunosuppressive pathways, including TLR signaling and myeloid chemotaxis.Functional pathway analysis confirmed a pronounced divergence between the two groups: responders engaged adaptive and effector immune programs, whereas non-responders displayed immunologically suppressed profiles.In conclusion, integrating clinical and single-cell data reveals that BsAb efficacy in RRMM is critically shaped by both tumor burden and baseline immune competence. These insights support the implementation of pre-treatment strategies aimed at lowering tumor burden and enhancing the effector-to-target ratio, thereby optimizing responses to bispecific antibody therapies.
INTEGRATED MULTIOMIC IMMUNE PROFILING DEFINES THE BIOLOGICAL BASIS OF MRD PERSISTENCE IN MULTIPLE MYELOMA Anna Maria Corsale Haematologica, 2026 Minimal residual disease (MRD) status is a strong predictor of outcome in multiple myeloma (MM), yet the immune mechanisms underlying MRD persistence or clearance remain unclear. To elucidate these mechanisms, we investigated the immunological correlates of MRD using an integrated multi-omic approach to dissect the cellular and functional remodeling of the bone marrow (BM) microenvironment after therapy.Flow cytometry was performed on BM aspirates from 23 ASCT-eligible MM patients (14 with undetectable MRD [uMRD] and 9 with persistent MRD [pMRD]) at baseline and day +100 post-ASCT. In eight patients (4 uMRD, 4 pMRD) with available baseline samples, additional 10-color panels were applied to achieve in-depth immune profiling of BM and peripheral blood. For seven patients (4 uMRD, 3 pMRD), baseline single-cell RNAseq integrated with TCR/BCR and 30-antibody CITE-seq was used to characterize the BM immune landscape.At baseline, pMRD patients exhibited lower frequencies of CD27⁻/CD28⁻/CD38⁻ T/NK cells compared with uMRD. At day +100, pMRD patients showed increased granulocytes, decreased lymphocytes, and higher granulocyte-to-lymphocyte ratio. A concomitant expansion of CD27⁺/CD28⁻/CD38⁻ T/NK cells was also noted. Extended immune profiling revealed that pMRD patients had reduced total T cells but an altered effector-to-naïve balance, with enrichment of CD8⁺ effector memory cells and depletion of CD8⁺ naïve cells in BM—mirrored in peripheral blood. Since no further differences were detected in terms of immune subset abundance, we next investigated qualitative transcriptional differences using a single-cell RNA sequencing approach. Baseline transcriptomic profiling revealed distinct immune programs associated with MRD status. In uMRD patients, CD8⁺ effector T cells showed enrichment in genes involved in adaptive immunity and memory maintenance pathways—including T-cell activation and differentiation, antigen processing and presentation, and memory formation—indicating a bone marrow milieu that supports sustained immune surveillance and coordinated B–T cell interactions. Similarly, CD14⁺ monocytes from uMRD patients displayed enrichment in activation and chemotaxis pathways (e.g., XYZ, SIAIHD, AISDSND), suggestive of enhanced support for T-cell–mediated immunity. Conversely, CD14⁺ monocytes from pMRD patients upregulated genes (e.g., XTW, ESD, FHAUIS) linked to humoral and innate immune responses, including antibacterial and antimicrobial peptide activity, B-cell activation and isotype switching, and toll-like receptor signaling. Finally, T-cell receptor (TCR) repertoire analysis demonstrated greater clonal variability and absence of dominant effector clones in pMRD patients at baseline, suggesting impaired immune coordination and reduced anti-myeloma activity. In conclusion, uMRD patients show a coordinated, therapy-enhanced immune microenvironment, whereas MRD persistence reflects immune dysfunction, suggesting targets to restore immune competence.
B Cell Levels in Centenarians, Semi-Supercentenarians, and Supercentenarians: Descriptive Analysis by Age, Sex, Cytomegalovirus Status, and Interleukin-6 Giorgio Bertolazzi, Anna Calabrò, Giulia Accardi, Anna Aiello, Calogero Caruso, et al. Journal of Ageing and Longevity, 2026 This study aims to characterise the B cell compartment in a cohort of Sicilian centenarians by analysing absolute CD3−CD19+ lymphocyte counts, in association with age, sex, cytomegalovirus (CMV) serostatus, related to immune ageing, and interleukin (IL)-6 levels, representative of inflamm-ageing. It also investigates age-related changes in the CD4+/CD19+ ratio as a marker of immune ageing, reflecting shifts in immune homeostasis. B cell counts were assessed by flow cytometry on 53 Sicilians aged 19–110 years: 20 Adults, 15 Older adults, 11 long-living individuals, and 7 oldest centenarians. A multiple negative binomial regression was applied to evaluate the effects of age, sex, CMV serostatus, and Il-6 levels on values of B cells. The results showed a non-significant trend toward age-related decline without sex-based differences. A significant reduction in B cell count was observed in individuals with high anti_CMV titres, while IL-6 levels showed a borderline inverse correlation. CD4+/CD19+ ratio values showed an age-related increase. Our findings suggest that the age-related decline in B cell numbers may be mostly related to CMV infection and IL-6 values, without sex contribution. The age-related increase in the CD4+/CD19+ ratio, most pronounced in oldest centenarians, may represent a compensatory adaptation promoting immune regulation and chronic inflammation control.
Identification of CD320, SLC44A1 and TNFRSF13B as potential novel therapeutic targets for CAR T-cell therapy in multiple myeloma Francesca Garofano, Anna Maria Corsale, Marta Biondo, Andrea Romano, Ingo Schmidt-Wolf, et al. Frontiers in Medicine, 2026 Introduction Multiple myeloma (MM) remains incurable despite effective therapies, with most patients eventually relapsing. Chimeric antigen receptor (CAR) T-cell therapy has improved treatment options but is limited by antigen escape and lack of durable responses. To expand the spectrum of actionable antigens, we sought to identify novel CAR-T actionable targets. Methods We analyzed 11 publicly available single-cell RNA sequencing datasets from patients with monoclonal gammopathy of undetermined significance (MGUS), smoldering MM (SMM), MM, relapsed/refractory MM (RRMM) and healthy donors. Plasma cell–specific surface proteins were identified via bioinformatic filtering and validated on external bulk transcriptomic database of ~2,000 patients and by flow cytometry on MM cell lines and primary bone marrow samples. Results We identified 15 plasma cell–associated surface proteins, including BCMA, CD138, CD38, and SLAMF7. Five molecules—TNFRSF13B (TACI), CD59, FCGR2B (CD32B), SLC44A1 (CD92), and CD320—were prioritized for further study. CD320 and SLC44A1 were upregulated with disease stage and associated with poor survival, while TNFRSF13B, CD59, and FCGR2B were enriched in advanced disease and linked to better outcomes. Cytogenetic clustering linked these molecules to specific genetic backgrounds, suggesting subtyperelated expression patterns. Flow cytometry confirmed the surface expression of CD59, SLC44A1, TNFRSF13B and CD320. Discussion CD320, SLC44A1, and TNFRSF13B are promising, clinically relevant targets for CAR T-cell therapy in MM. Their stage-specific expression and prognostic significance support their potential to enhance existing immunotherapeutic strategies.
Immune-Based Biomarkers as Predictors of Mortality in ECMO Therapy for Severe COVID-19 ARDS: Insights from a Retrospective Study Rosalia Busà, Giovanna Panarello, Alessia Gallo, Vitale Miceli, Salvatore Castelbuono, et al. International Journal of Molecular Sciences, 2026 Extracorporeal membrane oxygenation (ECMO) is a vital intervention for patients with severe respiratory failure, particularly in unresponsive acute respiratory distress syndrome (ARDS) cases. However, patient selection for ECMO remains a significant challenge. This study aims to identify novel immune-based biomarkers to improve eligibility assessment and predict outcomes in critically ill COVID-19 patients undergoing ECMO. This monocentric observational retrospective cohort study included 80 patients with severe COVID-19-related pneumonia who required ECMO support due to unresponsive ARDS. The patients were admitted to the intensive care unit (ICU) of IRCCS-ISMETT Hospital between September 2020 and April 2021, before the availability of COVID-19 vaccines. All patients were infected with the original SARS-CoV-2 Wuhan strain. Using machine learning approaches, the study analyzed clinical and laboratory data, cytokine levels, RNA sequencing (RNA-seq), and immune cell profiles collected within two days of hospitalization. The analysis identified a 5.56-fold increased mortality risk in patients presenting with a combination of immune factors: a T cell exhaustion profile, low interferon-alpha (IFNα) levels, and high calprotectin levels. These immune markers were strongly associated with poorer outcomes in patients undergoing ECMO. Our findings highlight the critical role of immune profiling in ECMO patient selection and outcome prediction. Incorporating immune-based biomarkers into clinical assessments may enhance the evaluation of ECMO eligibility and guide treatment decisions, ultimately improving patient outcomes.
BNT162b2 mRNA vaccination affects the gut microbiome composition of patients with follicular lymphoma and chronic lymphocytic leukemia Annalisa Chiarenza, Gaia Vertillo Aluisio, Nunziatina Laura Parrinello, Sara Marino, Anna Maria Corsale, et al. Biomarker Research, 2025 Background In both chronic lymphatic leukemia (CLL) and follicular lymphoma (FL) immunotherapy determines B-depletion that leads to temporary suppression of humoral immunity, which is clinically relevant especially during the COVID-19 pandemic, when most patients in the first wave received the BNT162b2 vaccine during anti-neoplastic treatment. Methods To capture changes in the immunome and microbiome composition in CLL and FL patients upon mRNA-based vaccination, we designed a prospective, longitudinal study to profile both the humoral and the cellular response after exposure to the BNT162b2 COVID-19 vaccine. Results In both CLL patients and FL patients, the second and third administrations of the BNT162b2 vaccine increased the titer of specific antibodies against SARS-CoV-2. In FL patients, vaccination induced expansion of central memory CD8 + CD57dim CD279 + T cells and reduction of the neutrophil subset myeloid 1 (CD14−CD15+CD16dimCD64+CD33−CD38+PDL1+HLA-DR−); in both cohorts, CD45RA + CD27 + CD279 + NK cells were expanded after a full cycle of vaccination. After vaccination, the genera Collinsella, Gemmiger, Lachnospiraceae, Blautia, Ruminococcus and Lactobacillus increased in both CLL patients and FL patients, whereas Faecalibacterium, Enterobacteriacae, and Enterococcus decreased. Multivariate analysis failed to identify factors associated with changes in microbiome communities among the CLL and FL cohorts, considering age, sex, exposure to anti-CD20 therapy and disease activity. Only in FL patients, alpha diversity was negatively correlated with neutrophil subsets myeloid 1 e 5 at baseline and positively correlated with neutrophil subset 6 after vaccination. PICRUSt2 analysis showed how microbiome can also affect the host health promoting chronic inflammation. The L-lysine biosynthesis pathway was more represented in CLL patients, whereas the L-valine degradation pathway and the anaerobic degradation of purine nucleobases were overrepresented in the FL cohort. Conclusions Taken together, our findings reveal the effect of the BNT162b2 vaccine in shaping the microbiome composition in CLL and FL patients, despite receiving treatment for their underlying active disease, and highlight the importance of a comprehensive analysis of the immunome and microbiome profiling to understand immune function in these cohorts of patients.
Inflammatory Biomarkers for Thrombotic Risk Assessment in Multiple Myeloma Patients on IMiD/aCD38-Based Regimens: Insights from a Prospective Observational Study Cirino Botta, Anna Maria Corsale, Claudia Cammarata, Fabiana Di Fazio, Emilia Gigliotta, et al. Biomolecules, 2025 Thrombosis is a common complication in multiple myeloma (MM) patients treated with immunomodulatory drugs (IMiDs), including thalidomide, lenalidomide, and pomalidomide. When combined with anti-CD38 monoclonal antibodies, these agents are highly effective but may increase thrombotic events (TE), potentially delaying therapy. This exploratory, hypothesis-generating analysis, conducted within the MMVision mono-institutional prospective study, included 53 MM patients who initiated IMiD plus anti-CD38 therapy between May 2021 and December 2022 (median follow-up: 18 months). Treatment regimens comprised lenalidomide (n = 36) or thalidomide (n = 15) with daratumumab, and pomalidomide (n = 2) with isatuximab. Most patients (n = 38) received frontline therapy, and all were given thromboprophylaxis according to guidelines, mainly aspirin (73%). Five patients (9.4%) developed VTE after a median of 48 days, managed with short-term low-molecular-weight heparin (LMWH). Exploratory analysis of 27 clinical/laboratory parameters suggested possible associations between VTE and low levels of beta-2 microglobulin, ferritin, intact/free lambda light chains, and monocyte-to-lymphocyte ratio. Notably, four of the five VTEs occurred in patients without lytic bone disease, typically associated with bone-driven inflammation in MM. Although all patients received aspirin prophylaxis from treatment initiation, it remains unclear whether thrombosis would also have occurred among those with higher inflammatory burden. These preliminary observations may indicate that in patients with relatively lower inflammation, aspirin prophylaxis could be less effective, potentially favoring VTE onset. In two VTE cases, cytokine profiling showed decreased M-CSF, SCLF-β, and MIP-1α, with increased G-CSF, raising the hypothesis of distinct immune-inflammatory pathways contributing to TEs. Given the limited number of patients and thrombotic events, and the cytokine data available for only two VTE cases, these associations should be regarded as exploratory and interpreted with caution. Overall, these exploratory findings warrant validation in larger, independent cohorts and may help generate hypotheses on how inflammatory signatures influence thrombotic risk and prophylaxis efficacy in MM patients receiving IMiD/anti-CD38-based regimens.
γδ T Cells in Glioblastoma Multiforme: Novel Roles and Therapeutic Opportunities Costanza Dieli, Rosario Maugeri, Anna Maria Corsale, Marta Di Simone, Claudia Avellone, et al. Cancers, 2025 Glioblastoma multiforme (GBM) is one of the most aggressive cancers, with limited treatment options due to its highly immunosuppressive microenvironment and resistance to conventional therapies. γδ T cells, known for their potent antitumor activity and ability to recognize tumor antigens independently of HLA molecules, have emerged as a promising therapeutic strategy. This review explores the role of γδ T cells in glioblastoma, focusing on their functional plasticity, cytotoxic mechanisms, and interactions with components of the tumor microenvironment. We examine the factors that influence γδ T cell polarization toward pro- or anti-tumor phenotypes and analyze preclinical findings that support their application in GBM treatment. Furthermore, we discuss potential combinatory approaches—including immune checkpoint inhibitors, cytokine stimulation, and adoptive cell transfer techniques—to enhance the therapeutic effectiveness and persistence of γδ T cells. Understanding the dynamics between GBM and γδ T cells may pave the way for innovative immunotherapeutic strategies aimed at overcoming immune evasion and improving clinical outcomes.
γδ T cell immunotherapy: Requirement for combinations? Anna Maria Corsale, Marta Di Simone, Francesco Dieli, Serena Meraviglia T Cell Cancer Immunotherapy Evidence Based Perspectives for Clinical Translation Volume 7, 2024
γδ cell-based immunotherapy for cancer Elena Lo Presti, Anna Maria Corsale, Francesco Dieli, Serena Meraviglia Expert Opinion on Biological Therapy, 2019
