Bronchoalveolar lavage fluid (BALF) is the most directly accessible sampling matrix for studying lower airway inflammation, yet conventional analyses rely on single-marker flow cytometry combined with analyte-by-analyte enzyme-linked immunosorbent assay (ELISA), an approach that limits information density and obscures cell-factor coupling. This study presents an integrated visualization workflow that couples standardized collection and preprocessing with a seven-marker neutrophil and monocyte flow cytometry panel, a 13-analyte multiplex cytokine assay, and a unified analysis pipeline based on uniform manifold approximation and projection (UMAP), FlowSOM metaclustering, hierarchical clustering, Spearman correlation networks, and principal component analysis (PCA). The workflow was applied to a single-center prospective cohort of 112 adults undergoing bronchoscopy, comprising 64 patients with microbiologically confirmed pulmonary infection and 48 non-infection controls. Cytological and flow cytometric readouts revealed a pronounced shift from a macrophage-dominant to a neutrophil-dominant alveolar profile during infection, with the neutrophil percentage rising from a median of 3.2% to 68.4% and the absolute neutrophil concentration increasing approximately 50-fold. Multiplex profiling resolved a coordinated three-module cytokine response, with chemokine, proinflammatory, and regulatory modules all upregulated and CXCL8/IL-8, IL-1β, IL-6, CXCL1/GRO-α, and G-CSF showing the strongest single-marker discrimination within the study cohort (area under the curve [AUC] 0.89 to 0.94). Integrative visualization using PCA, a correlation matrix, and a network graph revealed a tightly coupled neutrophil-chemokine axis at the participant level, with BALF neutrophil count emerging as the central network hub. The workflow uses only material routinely obtained from clinically indicated BAL, requires no additional sample volume, and provides an information-dense framework for resolving airway inflammation in respiratory infection that may be feasible in tertiary-care laboratories equipped with multiparameter flow cytometry and multiplex bead-array platforms, extending into longitudinal, single-cell, and therapeutic-evaluation studies. Broader clinical adoption of individual analytes as biomarkers will require prospective validation in independent multicenter cohorts.

Hematopoietic acute radiation syndrome (H-ARS) is an unmet medical requirement. Current cytokine-based countermeasures, such as granulocyte colony-stimulating factors, provide limited benefits once the surviving hematopoietic stem and progenitor cell pools are severely depleted. Toll-like receptor (TLR) agonists have been proposed as radiomitigators; however, their efficacy across ligands and receptor subtypes has not been systematically compared under equivalent conditions. Here we screened ten TLR agonists spanning eight receptor subtypes in a lethally irradiated mouse model and found that their radiomitigative efficacy was ligand-specific rather than receptor-class-specific. Monophosphoryl lipid A (MPLA), a detoxified TLR4 agonist, and FSL-1 (TLR2/6) exerted the strongest protective effects. MPLA conferred dose-dependent survival benefits when administered after lethal total-body irradiation, and this protection was substantially abolished by myeloid differentiation primary response 88 (MyD88) inhibition and partially reduced by competitive TLR4 blockade, suggesting that the TLR4-MyD88 signaling axis contributes substantially to the radiomitigative effect. Mechanistically, MPLA did not prevent the initial radiation-induced depletion of the bone marrow (BM), but markedly accelerated the subsequent regenerative phase, restoring multilineage peripheral blood counts, splenic and BM colony-forming activity, marrow architecture, and absolute numbers of hematopoietic stem/progenitor cell populations and mesenchymal stem/stromal cells. Regeneration was accompanied by selective induction of erythropoietin and thrombopoietin in the BM without broad changes in myeloid- or lymphoid-directed cytokines. Since MPLA already has an established human safety record as a licensed vaccine adjuvant, these findings support its repurposing as a potential radiomitigator for hematopoietic acute radiation syndrome.

Autologous stem cell transplantation (ASCT) following high dose chemotherapy is used for management of various pediatric malignancies, and plerixafor used in conjunction with GCSF is used to mobilize hematopoietic stem cells (HSCs) from bone marrow to peripheral blood for collection. While the efficacy and safety of plerixafor is established in adults, data for its use in children are limited. Factors influencing plerixafor response in pediatric patients have not been extensively investigated. This retrospective study aimed to identify predictors of response to plerixafor in pediatric patients undergoing ASCT. This single-center, retrospective study evaluated 22 pediatric patients who underwent mobilization with granulocyte colony-stimulating factor (GCSF) and plerixafor for ASCT between 2017 and 2024. Patient factors such as age, sex, baseline platelet count, marrow involvement, comorbidities, and malnutrition were recorded. Baseline platelet counts were measured on Day - 1 (the day prior to plerixafor administration). Treatment factors, including exposure to radiotherapy, anthracyclines, and multiple lines of chemotherapy, were also considered. CD34 + cell counts were measured before plerixafor administration and after harvest, and response was assessed by percentage, absolute, and log percent increase in CD34 counts. Regression analysis was done to look at the effect of various factors on response to plerixafor. The study found a significant increase in CD34 counts after plerixafor administration, with a median log percent increase of 3.54 (3.24-3.61). While age and malnutrition showed a significant correlation with log percent CD34 increase (p = 0.012 and p = 0.021, respectively), multivariate regression analysis revealed that only baseline platelet count achieved statistical significance as a predictor of response to plerixafor (p = 0.046). Pre-harvest baseline platelet count is an independent predictor of response to plerixafor in pediatric patients undergoing autologous stem cell transplantation. This finding suggests that baseline platelet counts can help predict the effectiveness of plerixafor-mediated stem cell mobilization in this patient population. Further larger prospective studies are warranted to validate these findings and identify additional factors influencing plerixafor response in pediatric patients.

Background and objectives Comprehensive immune-cell subset characterisation in healthy adults is the prerequisite for enabling accurate diagnosis of primary immunodeficiencies and informed therapeutic decision-making in targeted immunotherapies. Herein, we aim to establish biological reference intervals (BRIs) for leukocyte subsets for interpreting immunophenotypic data and identifying deviations in different pathological settings. Methods This prospective study was conducted at a tertiary care hospital and research centre in northern India over a period of six months using standardised multiparametric flow-cytometric immunophenotyping (FCMI). Peripheral blood from 120 healthy adult donors was analysed by 13-color, 15-parameter flow cytometry (Beckman Coulter Dx Flex). Thirty-seven leucocyte subsets were identified based on expression of surface markers including lymphoid (B, T and their subsets), myeloid (granulocytes, dendritic cells, monocytes and its subsets) and NK cell subsets. Frequencies and absolute counts were compared across gender and age groups by Mann-Whitney U and Kruskal-Wallis tests (P≤0.05). Results Males showed higher median total lymphocytes, absolute T-cells, cytotoxic T and memory T-regs, while females exhibited higher % of T-lymphocytes, helper T cells, and early terminally differentiated effector memory re-activating cells (TEMRA) fractions. B-cell analysis revealed higher mature, transitional, switched-memory, and circulating plasma-cell counts in males. Naïve B cells remained the predominant subset across adulthood, showing biphasic variation with age, while memory B cells peaked in early adulthood, followed by a decline. Interpretation and conclusions When compared with Euro-Flow data, Indian adults displayed overall concordance with global reference ranges, though with broader variability in T and B subsets. These baseline values will refine immune monitoring, improve diagnostic precision in immunodeficiency disorders and guide therapeutic decisions in immune-modifying therapies.

Hypereosinophilic syndrome (HES) is defined by persistent hypereosinophilia Q5 associated with eosinophil-mediated organ damage. However, the relationship between HES etiology and organ-involvement patterns remains incompletely defined, particularly in non-hematological forms. This study aimed to describe organ involvement across HES subtypes and to explore clinical associations with multiorgan disease.

Enfortumab vedotin (EV) is a crucial treatment for patients with metastatic urothelial carcinoma (mUC). However, a significant proportion of patients experience adverse events (AEs). The identification of biomarkers for AEs is imperative for the detection and treatment of AEs at an early stage. In this exploratory study, we aimed to identify biomarkers of AEs in patients with mUC treated with EV. We retrospectively examined 10 factors identified from the data of 116 patients with mUC treated with EV to identify biomarkers (age, body mass index, C-reactive protein level, Eastern Cooperative Oncology Group performance status, eosinophil proportion, history of diabetes, lymphocyte proportion, neutrophil proportion, neutrophil-to-lymphocyte ratio, and platelet count) associated with the occurrence of AEs of any grade. The candidate biomarkers were measured at the start of EV treatment. The least absolute shrinkage and selection operator method was used to select the most useful parameters for predicting AE occurrence. Among the 10 factors, eosinophil proportion was identified as the only potential biomarker. The optimal cutoff value for eosinophil proportion against the occurrence of AEs of any grade was 2.5% (area under the curve = 0.625). Univariable logistic regression analyses showed that an eosinophil proportion of ≥ 2.5% was a risk factor for AE development (odds ratio = 4.35, 95% confidence interval = 1.35-14.0). Therefore, the results of this exploratory study indicated that an eosinophil proportion of ≥ 2.5% at the start of EV treatment may be a candidate biomarker for the occurrence of AEs of any grade.

Autoimmune neutropenia (AIN) in early childhood is caused by autoantibodies targeting neutrophil surface antigens, most commonly CD16 (FCGR3B), and must be distinguished from congenital neutropenia, malignancy-associated neutropenia and chronic idiopathic neutropenia (CIN). In this study, the diagnostic performance of the cell-based granulocyte agglutination test (GAT) and granulocyte immunofluorescence test (GIFT) was compared with the bead-based LabScreen Multi (LSM) assay in 214-suspected AIN cases. While 36% of samples were positive for anti-CD16 antibodies in GAT or GIFT, only 19% reacted against the same antigen in LSM, primarily against weak or Immunoglobulin M (IgM)-restricted antibody responses. Patients testing positive in all three assays exhibited significantly higher pro-LL-37 levels and lower absolute neutrophil counts (ANCs), indicating preserved granulopoiesis with peripheral destruction. Our findings question the use of LSM as a standalone test in AIN diagnostics. Incorporating pro-LL-37 and ANC in AIN diagnosis may improve differentiation between antibody-mediated neutropenia and other neutropenia forms in children.