The field of immunotherapy for unresectable soft tissue sarcomas (STS) is transitioning from a one-size-fits-all approach toward a precision immuno-oncology paradigm. The pronounced immunological heterogeneity among histological subtypes and the limited predictive capacity of traditional classification underscore the imperative for immune-based stratification. Approximately 20% of STS exhibit an immune-activated tumor microenvironment, characterized by robust cytotoxic T lymphocyte infiltration, B-cell enrichment, and tertiary lymphoid structures (TLS); these patients demonstrate significantly higher objective response rates to immune checkpoint inhibitors (ICIs) and may be prioritized for such therapy. TLS status could be incorporated into routine clinical decision-making as a robust immunological biomarker for treatment selection. For the majority of patients with TLS-negative, immunologically "cold" tumors, however, single-agent ICIs are insufficient. Combination strategies designed to remodel the immunosuppressive tumor microenvironment represent a promising approach: enhancing tumor immunogenicity through epigenetic modulators, improving antigen presentation via CD47/SIRPα blockade, and exploring dual-checkpoint blockade to overcome T-cell exhaustion. For translocation-associated sarcomas, where neoantigen generation is inherently limited, adoptive cell therapies targeting specific antigens represent a particularly promising avenue. Biomarker-driven basket or umbrella trial designs are paramount to efficiently identifying optimal combination regimens and improving overall survival outcomes for patients with unresectable disease.
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Progress in immuno-oncology has been stymied by poorly immunogenic 'cold' tumours and the focus on T cells at the cost of other immune cells. Here we report that AT-1965, a small molecule in a lipid nanoparticle, induces rapid regression of poorly immunogenic tumours with the formation of immune memory through interaction with Cap-specific RNA (nucleoside-2'-O-)-methyltransferase 2 (CMTR2) in cancer cells, triggering an innate inflammatory viral defence response. AT-1965-treated tumours were found to be highly infiltrated with B cells, which are known to act as early responders to viral signatures. Genetic knockout of functional B cells abrogated the anti-tumour efficacy of AT-1965, directly implicating B cells in the anti-tumour outcome. Our results rationalize clinical data showing that patients with high CMTR2 expression in tumours have a poor prognosis and that B cell infiltration is associated with long-term survival in multiple tumour types. The AT-1965 nanomedicine-inspired discovery of CMTR2 as a potential cancer target and B cell recruitment opens a new frontier for immuno-oncology.
Boron neutron capture therapy (BNCT) is a targeted radiotherapeutic modality that employs boron-10 (10B) to capture thermal neutrons, thereby releasing high-linear energy transfer α-particles and lithium ions that selectively eradicate tumor cells while sparing the surrounding normal tissues. Immune checkpoint inhibitors (ICIs), including inhibitors of PD-1/PD-L1 and CTLA-4, enhance antitumor immunity by alleviating immunosuppression within the tumor microenvironment. As immunotherapy continues to expand across multiple tumor types and becomes integral to systemic cancer treatment, attention has increasingly focused on combining systemic immunomodulation with localized therapies. BNCT functions primarily as a local therapeutic approach, whereas ICIs elicit systemic immune activation. Whether the two modalities can produce synergistic therapeutic effects remains an important question.In this narrative review, we summarize current mechanistic insights, preclinical and clinical developments, and ongoing challenges associated with the combination of BNCT and ICIs, highlighting the potential of this strategy to achieve durable, synergistic antitumor responses.
Inflammation in the female genital tract (FGT) is a key risk factor for HIV acquisition, but it remains unclear whether clinical or immunological measures best predict risk. We aimed to prospectively compare HIV acquisition among women with clinically and/or immunologically defined inflammation. HIV-uninfected women enrolled in the CAPRISA 004 tenofovir gel randomized controlled trial in South Africa were followed for up to 34 months. We analyzed data from 889 women, with cytokine measurements available for 774 participants. Clinical genital abnormalities were assessed at scheduled visits, and 48 cytokines were measured in cervicovaginal lavage samples. HIV incidence was compared across categories of clinical and immunological inflammation using time varying Cox proportional hazards models, adjusting for relevant covariates. Immunological inflammation, defined as ≥9 elevated cytokines, was present in 18% (140/774) of women. Among specific clinical signs, abnormal genital discharge (aHR: 2.67, 95% CI: 1.14-6.23, p=0.024) and cervicitis (aHR: 10.34, 95% CI: 2.46-43.65, p=0.001) were significantly associated with increased HIV acquisition. Women with both clinical and immunological inflammation had the highest risk of HIV acquisition, with adjusted hazard ratios of 2.08 (95% CI: 1.10-3.91, p=0.022) and 2.46 (95% CI: 1.21-5.03, p=0.013), respectively. Clinical and immunological definitions of inflammation were each independently associated with increased HIV acquisition risk and combined they reflected greater susceptibility. These findings highlight the important role of genital inflammation in women's HIV susceptibility, suggesting that clinical signs may provide practical early indicators of risk even as cytokine profiles provide a more sensitive measure of underlying inflammation.
Tumour-draining lymph nodes (TDLNs) serve as the closest immunological hubs to the primary tumour site in colorectal cancer (CRC). Owing to their unique and irreplaceable anatomical advantage and dynamic immune cell repertoire, TDLNs represent an intensively studied immunological niche and are a potential therapeutic target. This study aimed to map the immune microenvironment of CRC TDLNs to identify targetable immunomodulatory axes. Single-cell RNA sequencing (scRNA-seq) was performed on 23 quadruplet-matched samples, including primary tumours, adjacent normal tissues, tumour-free lymph nodes (TFLN) and tumour-invaded lymph nodes (TILN) from seven CRC patients. Mechanistically significant findings were further validated through in vitro functional assays, CRISPR knockout in primary regulatory T cells (Tregs), in vivo murine footpad-popliteal lymph node metastasis models with lipid nanoparticle-encapsulated siSPP1 (LNP-siSPP1) and/or anti-CD44 mAb, and multiomics analysis of independent CRC cohorts. scRNA-seq analysis delineated TILN-specific immunological landscapes dominated by SPP1+ macrophage expansion and active Treg differentiation niches, establishing TILNs as maturation hubs for Tregs versus TFLNs. Mechanistically, SPP1+ macrophages drove Treg differentiation into immunosuppressive CD137+ subsets via the SPP1-CD44 axis, which required NF-κB1 to directly bind the TNFRSF9 promoter. In vivo, LNP-siSPP1 plus anti-CD44 mAb synergistically suppressed lymph node metastasis, reduced CD137+ Tregs and enhanced CD8+ T cell function. Findings were consistently observed across all experimental models and patient-derived datasets. SPP1+ macrophages established an immunosuppressive niche in CRC TDLNs by promoting CD137+ Treg maturation via the SPP1-CD44-NF-κB1 axis. Targeting this axis with LNP-siSPP1 and anti-CD44 mAb might overcome Treg-mediated immunosuppression in CRC.
Autologous tumor vaccines have been investigated as immunotherapeutic strategies in veterinary oncology and may represent relevant translational models for comparative cancer immunotherapy. However, reported clinical outcomes in dogs remain heterogeneous. To evaluate the safety, clinical outcomes, and immunological assessment methods of autologous tumor vaccines in dogs. A systematic review of PubMed, Web of Science, and Scopus identified 24 eligible studies evaluating autologous tumor vaccines in dogs. Vaccine platforms, safety data, clinical outcomes, and immunological assessments were analyzed descriptively. Exploratory pooled hazard ratio analyses were performed using random-effects models when applicable. Autologous tumor vaccines were generally reported as well tolerated, with predominantly mild and transient adverse events. Clinical outcomes varied substantially across tumor types and study designs. Canine lymphoma represented the setting with the greatest concentration of evidence, including the few randomized and controlled studies identified. Several lymphoma studies reported longer time to progression and lymphoma-specific survival in vaccinated dogs compared with chemotherapy-alone or historical comparator groups, although findings were not consistent across all studies. Exploratory pooled analyses suggested a trend toward reduced risk of progression and lymphoma-related mortality in immunotherapy-treated dogs; however, these findings should be interpreted cautiously given the limited number of studies and methodological heterogeneity. In solid tumors, outcomes were more variable and evidence remained predominantly exploratory. Current evidence suggests that autologous tumor vaccines in dogs are generally feasible and well tolerated, although methodological limitations restrict interpretation of therapeutic efficacy. Further prospective and standardized studies are needed to clarify their role in veterinary oncology and comparative cancer immunotherapy.
Psoriatic arthritis (PsA) often requires escalation from conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) to biologic therapy (bDMARDs), yet biomarkers guiding treatment decisions remain limited. Anti-CD74 autoantibodies have shown diagnostic potential in axial spondyloarthritis, but their relevance in PsA is insufficiently characterized. To evaluate whether IgA anti-CD74 levels are associated with treatment escalation in peripheral PsA (pPsA) and to characterize their relationships with clinical and immunological parameters. Serum samples from 171 PsA (127 pPsA, 44 axial PsA [axPsA]), 43 non-rheumatic disease controls (NRD), and 43 rheumatoid arthritis (RA) patients were analyzed. IgA anti-CD74 levels were measured by enzyme-linked immunosorbent assay (ELISA). Patients were stratified by disease duration and treatment exposure. Correlation analyses, receiver operating characteristic (ROC) curves, and logistic regression models were performed. A prospective subgroup of 53 early pPsA patients was followed to assess treatment initiation. IgA anti-CD74 levels were elevated in PsA compared with NRD controls (median 13 vs. 6 U/mL, p < 0.0001), with similar levels in pPsA and axPsA and comparable values in RA, indicating an association with inflammatory disease rather than disease specificity. Anti-CD74 positivity (>15 U/mL) was observed in 31.5% of pPsA and 43.2% of axPsA versus 2.3% of NRD, independent of disease duration. Anti-CD74 levels were associated with treatment escalation in pPsA, with higher levels in bDMARD-treated patients (median 13.0 vs. 11.0 U/mL, p = 0.04). In multivariate analyses, anti-CD74 was independently associated with csDMARD (OR 1.113, p = 0.022) and bDMARD use (OR 1.052, p = 0.02). After false discovery rate (FDR) correction, anti-CD74 remained associated with serum IgA (q = 0.0008) and weakly with IgG (q = 0.0250), but not with C-reactive protein (CRP) or age. Longitudinal associations were not significant after FDR correction (csDMARD initiation: p = 0.047, q = 0.094; bDMARD initiation: p = 0.19, q = 0.1866), indicating these findings are exploratory. IgA anti-CD74 levels are elevated in PsA and appear to reflect immunological activity not captured by CRP. Their independent association with treatment escalation in pPsA supports further evaluation as a biomarker candidate, although findings remain exploratory and require validation in larger longitudinal cohorts.
Osteoarthritis is the predominant joint ailment. Multiple studies demonstrate that the dysregulation of catalytic regulators of ubiquitination and deubiquitination disrupts cartilage imbalance, consequently facilitating the advancement of osteoarthritis. Ubiquitination-related biomarkers for osteoarthritis were found by differential expression, weighted gene co-expression network analysis, Mendelian randomization, Receiver Operating Characteristic curves, and expression analyses. Subsequently, an examination of immune infiltration was conducted to evaluate the contrasting immunological circumstances between osteoarthritis and controls. Additionally, single-cell analysis was employed to screen key cell types, and analyze the expression of biomarkers during their differentiation. The expression of biomarkers was subsequently validated using real time quantitative polymerase chain reaction. CBLB and NQO2 were determined as biomarkers, having risk effects on osteoarthritis (odd ratio > 1). Analysis of immune infiltration indicated a significant disparity in the number of 15 immune cell types between osteoarthritis and control groups, such as type 2 T helper cells and macrophages, and two biomarkers showed opposite associations with these immune cells. Single-cell analysis annotated seven cell types, with prehypertrophic chondrocytes as the key cells. Notably, two biomarkers had expression early and late stages during prehypertrophic chondrocytes differentiation. Finally, experiments analysis indicated that CBLB decreased and NQO2 increased in osteoarthritis samples. CBLB and NQO2 were biomarkers associated with ubiquitination that exert causal effects on osteoarthritis. These findings provide potential therapeutic targets for clinical intervention and help to personalize treatment for osteoarthritis patients.
The Danish Database for Chronic Hepatitis B and C (DANHEP) was established in 2002 to support research into the clinical, epidemiological, virological, and immunological aspects of chronic viral hepatitis in Denmark. The objective of this paper is to describe methods of data acquisition, content, completeness and accuracy of data in DANHEP. Since 2002 all individuals with chronic viral hepatitis B or C at least 16 years of age, who has presented to a hospital department specialized in infectious diseases or gastroenterology in Denmark, have been enrolled in DANHEP. Information on co-infections, viral DNA/RNA detection, serology, liver parameters, liver fibrosis, treatment initiation, -cessation and -efficacy is manually transferred from electronic patient files and recorded in DANHEP annually. In 2004, a biobank storing annual blood samples was established. By December 2023 DANHEP stored data on 15,886 individuals with chronic hepatitis B or C and the biobank held approximately 38,180 samples. We verified a random sample of key variables by comparing with electronic patient files and found an error proportion of less than 5%. DANHEP is well suited for studying the epidemiology, virology and immunology of chronic viral hepatitis. We encourage future collaborations to address the many remaining questions in the field of viral hepatitis.
Allergic sensitisation is traditionally viewed as a stable trait despite studies suggesting that it may vary across life-course. This study aims to provide longitudinal evidence of allergic sensitisation and its associated factors in the general population. Aeroallergen sensitisation was assessed in the Austrian population-based cohort using repeated skin prick testing (SPT) across 3 visits (mean visit interval, 4.25 ± 0.33 years). Longitudinal sensitisation patterns were characterised as stable nonsensitised, stable sensitised, new-onset, resolution or fluctuating. We examined demographic, metabolic, behavioural, environmental and immunological factors across age strata (< 18, 18 ≤ 40, 40 ≤ and ≥ 60 years). Among 5046 individuals with follow-up and valid SPT in all 3 visits, 54.4% remained stably nonsensitised and 30.6% stably sensitised, 5.6% experienced resolution, 4.8% new onset and 4.6% fluctuating sensitisation. New onset expectedly predominated in < 18 years, fluctuation in 18 ≤ 60 years and resolution in ≥ 60 years, indicating that sensitisation is modifiable and time-varying. Stable sensitisation was strongly associated with parental allergy and blood eosinophils while cumulative smoking modestly reduced its likelihood. Fluctuation was linked to adiposity (in adults ≥ 40 years old) and environmental exposures. Aeroallergen-specific analysis showed new-onset sensitisation was driven by outdoor allergens (ragweed, tree pollens and pets) whereas fluctuation was driven by seasonal pollens. Although predominantly stable, allergic sensitisation is dynamic and exhibits substantial temporal variation. Resolution and fluctuation occur across all ages and new sensitisation can emerge well into adulthood. These findings challenge the traditional view of fixed sensitisation and suggest it reflects a modifiable phenotype shaped by age and environmental and lifestyle exposures.
Neuroimmune communication is essential for regulating inflammation and maintaining cardiovascular homeostasis, but the role of sensory pathways in this process is poorly understood. Arterial baroreceptors are typically defined as mechanoreceptors essential for arterial pressure homeostasis and have been associated with modulation of the immune response. However, their role in sensing systemic inflammation remains unknown. Here, we establish the molecular profile of the rat aortic depressor nerve (ADN) as an immune-competent tissue and investigate its response to lipopolysaccharide (LPS)-induced endotoxemia. Using analysis of gene expression, total protein quantification, and immunofluorescence assay, we demonstrate that the ADN, from male Sprague-Dawley rats (7-8 weeks old), constitutively expresses key components for innate immune signalling, including Toll-like receptor 4 (TLR4), MyD88, and phosphorylated NF-κB, indicating a state of constant immunological vigilance. LPS administration induced an inflammatory response within the ADN, upregulating gene expression of NF-κB, interleukin-6, and type I interleukin 1 receptor, and it also increased the ADN electrical activity. Notably, the increase in nerve firing occurred while the animals were experiencing systemic hypotension and also during the diastolic phase, indicating that this response is not from the mechanosensory reflex. Furthermore, we characterized the progression of this immune response in the nodose ganglion and aortic arch, identifying a coordinated neuroimmune sensory axis. These findings reposition arterial baroreceptors from purely mechanoreceptors to integrative immunosensors that actively detect and respond to systemic inflammation. This novel neuroimmune circuit represents a critical link between inflammation and cardiovascular system, offering a novel therapeutic target for treating cardiovascular and inflammatory conditions.
Chimeric antigen receptor T-cell (CAR-T) therapy was initially used to treat B-cell malignancies, and it is now considered an effective treatment option for multiple sclerosis (MS). CAR-T therapy selectively targets and depletes pathogenic B cells within lymphoid tissue and the central nervous system (CNS), showing promise for achieving deep, sustained remission and long-term treatment-free disease control in patients with refractory MS. A comprehensive analysis was carried out by searching multiple keywords with combinations such as "CAR-T", "MS", "Demyelination", "Autoimmunity", "CD19", "Inflammation", "B cells", T cells", "Neurodegeneration ", "Neurological Disorders", "Immunity", etc. The review included preclinical and clinical research articles publicly available till March 2026. This study was conducted to explore the mechanisms of action, clinical effectiveness, safety profile, and prospects for CAR-T treatment for MS. From the beginning clinical testing indicates that CD19 targeted CAR-T cells can efficiently and permanently destroy through B-cells, leading to a significant decrease in disease progression, a recovery of impairment, and an immense reduction in inflammatory markers in individuals who have progressive MS. New techniques for engineering such as allogeneic CAR-T cells and enhanced CRISPR-based safety switches, are being investigated for making things safer and easier for individuals. CAR-T treatment represents a revolutionary approach for individuals with refractory MS. With ongoing improvements in safety and specificity, it has the potential to transform the therapeutic paradigm toward a sustainable immunological reset and prolonged remission in clinical neuroimmunology.
Cell migration across epithelial barriers occurs in diverse developmental, immunological, and pathological contexts. Here, we investigate the contribution of heterotypic adhesion between migrating cells and epithelial "substrate" cells to transepithelial migration. Using an in silico model inspired by the migration of primordial germ cells across the midgut epithelium in the Drosophila embryo, we show that heterotypic adhesion modulates migration efficiency in a nonmonotonic manner, revealing the existence of an optimal adhesion regime. Consistent with this prediction, in vivo overexpression of E-cadherin in germ cells accelerated their exit from the midgut relative to controls. Beyond providing experimentally testable predictions, our model integrates and explains previous observations on the role of heterotypic adhesion in cell-on-cell migration, offering a framework for understanding transepithelial migration across biological contexts.
Hydrosalpinx is a chronic pathological manifestation seen in approximately 30% of tubal disease cases. In addition to contributing to infertility and pelvic pain, hydrosalpinx has been associated with early pregnancy loss and reduced implantation in assisted reproductive technology. Though not fully understood, previous studies have demonstrated altered immunological activity in those with hydrosalpinx, which has been suggested to impact endometrial receptivity. To validate these findings and to explore the pathogenesis of tubal disease, the aim of this study was to identify and quantify the immune cell population in tubes with hydrosalpinx and endometrium from the same patients. This observational study included patients in the secretory phase of the menstrual cycle undergoing salpingectomy/salpingo-oophorectomy with/without hysterectomy for benign gynaecological conditions. The number and type of immune cells present in endometrial and fallopian tube tissue sections were investigated using immunostaining for CD45, CD3, CD4, CD8, CD56 and CD68. We show that T cells are more abundant in the endometrium of women with hydrosalpinx and that cytotoxic T cell numbers are lower in diseased tubes. Uterine natural killer cells and macrophages are significantly elevated in both tubal and endometrial tissue in hydrosalpinx, demonstrating a disruption in the normal uterine immune cell microenvironment that may affect endometrial receptivity, with implications for tubal disease-associated subfertility. Hydrosalpinx is a common condition characterised by inflammation, distortion and fluid accumulation in the fallopian tube. Hydrosalpinx is a leading cause of subfertility, however, the mechanisms that cause this problem are not fully understood. We studied the immune cells present in both the fallopian tubes and uterine lining (endometrium) of women with hydrosalpinx and compared them to samples from women without tubal disease. The samples were collected in the second half of the menstrual cycle, when the uterine lining is prepared for implantation of a fertilised embryo. We quantified the total immune cell numbers and specific immune cell subtypes (these are immune cells with specific roles in the body, such as destroying harmful bacteria or regulating the immune system) in the different tissues and compared them between the hydrosalpinx and control groups. We found that hydrosalpinx is associated with both tubal and endometrial immune cell changes, which may contribute to infertility by affecting the normal physiological processes that allow the endometrium to accept an embryo and establish a pregnancy.
Tularemia, caused by Francisella tularensis, is a zoonotic disease with various sources, transmission routes, and geographically different clinical signs. Diagnosis is challenging due to nonspecific symptoms, highlighting the importance of laboratory testing for accurate detection, effective outbreak management, and targeted treatment. Recent improvements have enhanced the accuracy of sensitive and specific immunological and molecular methods. Techniques such as enzyme-linked immunosorbent assay, immunochromatography, microagglutination tests, indirect immunofluorescence assays, and PCR continue to be key detection tools. Molecular typing methods such as whole-genome sequencing, single-nucleotide polymorphism analysis, multiple-locus variable-number tandem repeat analysis, pulsed-field gel electrophoresis, and matrix-assisted laser desorption ionization-time of flight enable precise genetic characterization of F. tularensis strains. These methods improve understanding of phylogeny, strain diversity, and transmission routes, while supporting outbreak investigations. This review aims to provide a comprehensive overview of current laboratory methods for diagnosing and typing tularemia as well as their application in clinical and research settings.
Children with Down syndrome (DS) have a high prevalence of obstructive sleep apnea (OSA) due to anatomic, neuromuscular, immunological and metabolic factors, yet the contribution of the tonsillar microbiome to airway obstruction in this population remains unexplored. We hypothesized that DS-associated OSA would be associated with a distinct tonsillar microbiome compared to non-DS OSA. Tonsillar tissue from 22 DS and 18 NDS participants were analyzed by 16S rRNA sequencing. Alpha and beta diversity were assessed using Faith's phylogenetic diversity and UniFrac distances, respectively, and significantly different taxa were identified with ANCOM-BC and Mann-Whitney testing. Although overall microbial richness and community structure were similar between groups, overweight DS participants demonstrated increased phylogenetic diversity compared to normal-weight DS peers. Taxonomic profiling of the entire patient cohort revealed that in DS tonsils there were selective alterations in key genera with selective depletion of Haemophilus and enrichment of Staphylococcus , Rothia , and Lactobacillales . Haemophilus abundance correlated positively with tonsil weight in both cohorts. These findings suggest that while global diversity is preserved, specific microbial shifts distinguish the DS tonsillar niche, potentially reflecting altered immune and metabolic environments associated with trisomy 21. Understanding these microbial differences may reveal mechanisms underlying the higher incidence and persistence of OSA in DS and inform targeted therapeutic strategies.
Based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO), we profiled the role of Colony-Stimulating Factor 1 Receptor (CSF1R) in cancer. We specifically focused on pancreatic ductal adenocarcinoma (PAAD), analysing its regulative function in cancer progression and its usefulness as a potential biomarker and therapeutic target. Using the TCGA and GEO databases, gene expression, survival prognosis, genetic abnormalities, immune infiltration, and CSF1R-related gene enrichment related to the CSF1R gene in patients with PAAD were studied. CSF1R expression appears to affect the development of PAAD; there is more CSF1R expression in the tumour tissue than in nearby normal tissues. There was no statistical difference in the overall survival or diseasefree survival of CSF1R expression. Analysis of changes in the CSF1R gene showed that PAAD samples had a low mutation frequency, and CSF1R gene amplification was the main reason. It was additionally emphasized that the impact of CSF1R on the tumor microenvironment, as evidenced by an immunological infiltration analysis, showed a strong correlation between the estimated infiltration values of cancer-associated fibroblasts and CSF1R expression in PAAD. This additional evidence was found for CSF1R expression in cancer-associated fibroblasts in PAAD. Targeting CSF1R might be a promising strategy for PAAD treatment. Inhibiting CSF1R activity or TGF-β binding to CSF1R inhibits tumour growth and immune escape. Investigating the link between CSF1R, TGF-β, and the immune system leads to new opportunities for combination therapies that integrate targeted medicine with immunotherapy. Targeting CSF1R might be a PAAD treatment. Inhibiting CSF1R activity or TGF-β binding to CSF1R inhibits tumour growth and immune escape. Investigating the link between CSF1R, TGF-β, and the immune system opens new opportunities for combining targeted medicines with immunotherapy.
A performance evaluation of a quantitative research use only (RUO) immunological assay utilizing the Meso Scale Discovery (MSD) platform for assessing IgM, IgA, and IgG binding and SARS-CoV-2 pseudo-neutralization was performed. Comparative analyses of antibody quantification of IgA, IgG, and IgM isotypes against SARS-CoV-2 nucleocapsid, spike, Receptor Binding Domain (RBD) and N-Terminal Domain (NTD) antigens were performed using MSD multi-spot electro-chemiluminescent V-PLEX COVID-19 test kits, and two well characterized serological panels. Cross reactivity was assessed against COVID-19 negative/Influenza samples. Precision was assessed by comparing variation between duplicate values within run; accuracy was assessed by comparing values obtained across runs against expected values. The MSD IgG, IgA, and IgM binding, and ACE2 inhibitory antibody assays demonstrated acceptable precision and accuracy within the expected 20%/30% variability, respectively per manufacturer's protocol; no cross reactivity with influenza samples was observed. Different MSD plate configurations, multiple (two) MSD analysis instruments, and multiple operators did not impact antibody quantification; thus, confirming the robustness of both assays. Comparative analysis between MSD and commercial Euroimmun, Siemens ADVIA, and Beckman Coulter DXLTM ACCESS assays demonstrated better performance overall for the MSD assay. IMPORTANCE: This work describes and validates the MSD platform, as a highly flexible, customizable, operator independent, and reliable platform for antibody assessment of COVID-19 response which is highly adaptable to immune assessment of emerging or re-emerging pathogens.
Osteoarticular tuberculosis (OATB) is one of the most common clinical types of extrapulmonary tuberculosis (EPTB), characterized primarily by progressive bone destruction and high morbidity. Standard anti-tuberculous chemotherapy has limited efficacy in reversing established bone damage. Vitamin D, a fat-soluble steroid hormone with immunomodulatory and bone-metabolic properties, shows promise as an adjunctive therapeutic agent. Mechanistically, vitamin D enhances anti-tuberculous immunity through multiple pathways: inducing antimicrobial peptide LL-37 expression, activating autophagic flux, and modulating Th1/Th17/regulatory T cell balance. At the bone level, vitamin D counteracts Mtb-induced destruction via the OPG/RANKL axis, Wnt/β-catenin signaling activation, and correction of secondary hyperparathyroidism. However, rifampicin-a cornerstone chemotherapy agent-accelerates vitamin D catabolism by inducing CYP3A4/CYP24A1, creating a clinically significant drug-nutrient interaction that exacerbates vitamin D depletion. Evidence supporting vitamin D supplementation remains predominantly derived from pulmonary tuberculosis (PTB) patients in randomized controlled trials (RCTs), with only one small-sample study specific to OATB (n = 41, 8 weeks). These PTB-derived trials employed single high-dose bolus regimens with 3-6 months follow-up, demonstrating modest benefits (HR 0.58-0.89) primarily in vitamin D-deficient subgroups, with negligible effects in replete populations. Direct applicability to OATB remains unverified. This review systematizes vitamin D's immunological and bone-metabolic mechanisms in OATB, critically appraises existing evidence quality and extrapolation limitations, and proposes an RCT framework with bone structural repair and functional recovery as primary endpoints. Our aim is to guide theoretical development and future clinical investigation of vitamin D as adjunctive therapy for OATB.