Search Results (8,905)

Background: The COVID-19 pandemic exposed critical gaps in pandemic preparedness and highlighted the need for vaccine platforms capable of rapid adaptation. Outer membrane vesicle (OMV)-based platforms utilizing vesicles derived from genetically detoxified Neisseria meningitidis serogroup B (Nm-nOMV) represent a promising plug-and-play approach. Methods: This Phase I, first-in-human, randomized, double-blind, placebo- and OMV-controlled trial, evaluated safety, tolerability, and immunogenicity of intranasally administered OMVs combined with SARS-CoV-2 Spike protein in healthy SARS-CoV-2 seropositive adults aged 18–55 years. Forty participants were enrolled across two cohorts: a low-dose cohort receiving 140 μg OMV/70 μg Spike (OMV + Spike, n = 13; OMV alone, n= 3; Placebo, n = 5) and a high-dose cohort receiving 280 μg of OMV/140 μg of Spike (OMV + Spike, n = 13; OMV alone, n = 3; Placebo, n = 3), administered on Days 1 and 22. Safety was assessed through adverse events, vital signs, laboratory parameters, ECG, and pulse oximetry. Immunogenicity was evaluated via systemic SARS-CoV-2 neutralizing antibodies, antigen-specific antibodies (IgG and IgA), and mucosal antibodies (IgA in nasal wash). Results: Intranasal administration of OMVs combined with SARS-CoV-2 Spike protein was safe, well-tolerated, and immunogenic. No serious adverse events were reported, and adverse events were predominantly mild and transient. Dose-dependent increases in systemic and mucosal immune responses were observed, with statistically significant enhanced serum IgG and nasal wash IgA antibodies in the high-dose group. Conclusions: The current clinical data confirm key aspects of the preclinical profile, which demonstrate the potential of the Nm-nOMV platform as a strong adjuvant for mucosal vaccines. These findings support the broader application of the Nm-nOMV vaccine platform in pandemic preparedness. Full article

Background/Objectives: For low-grade glioma (LGG) patients, adjuvant treatment (AT) with radiotherapy and chemotherapy may adversely impact cognition. However, existing evidence is limited by methodological heterogeneity and shortcomings. This study explored the cognitive functioning of LGG patients who underwent resection with both radiotherapy and chemotherapy (AT+) or without AT (AT−), from before resection to one year after resection. Methods: We included patients with World Health Organization 2021 grade 2 isocitrate dehydrogenase-mutated glioma who underwent resection between 2011 and 2024. All patients completed a neuropsychological screening battery one week before (T0) and twelve months after resection (T12), measuring reaction time, attention span, information processing speed, working memory, inhibition, cognitive flexibility, and verbal fluency. We compared cognitive performance between AT+ and AT− patients at T0 and T12, as well as trajectories of cognitive functioning, at the group and individual level. Results: We included 60 LGG patients (M age = 38.8 years; 63.3% male). Compared to AT− patients (n = 35), AT+ patients (n = 25) were significantly older, more frequently had tumors that crossed the midline, and reported more depressive symptoms. At T0, no significant cognitive performance differences existed between AT+ and AT− patients, despite lower observed performance in the AT+ group. At T12, AT+ patients performed significantly worse than AT− patients on mean information processing speed, due to an improvement over time in the AT− group. Conclusions: Patients allocated to AT may show limited cognitive recovery of information processing speed up to 12 months after surgery, without pronounced effects on other cognitive functions. These findings can guide future studies into treatment-related cognitive decline of LGG patients. Full article

Background and Objectives: Up to 40% of patients with cancer develop spinal metastases, and stereotactic spinal radiosurgery (SSRS) achieves high local control rates as definitive or postoperative treatment. Multiple tumor response assessments have been used but their compared clinical performance in post-surgical patients remains unclear. We sought to compare the applicability of RECIST1.1, MDACC and SPINO criteria. Materials and Methods: This IRB-approved retrospective study included patients with high-grade epidural spinal cord compression treated with decompressive surgery followed by adjuvant SSRS, with MRI follow-up available. Lesions were classified according to each of the scale’s objective (RECIST1.1 and MDACC) and subjective (SPINO [radiology reports]) criteria. Results: Ninety-four treated levels in 93 patients (median age 58.9 years) were analyzed. Most metastases were thoracic, and all cases had preoperative high-grade epidural spinal cord compression. Adjuvant SSRS was delivered in one or three fractions. Median follow-up was 16 months (range, 1–132), SPINO-based assessment was feasible in 100% of cases, RECIST1.1 in 43.6% and MDACC in 46.8%. Progressive disease criteria were met in 21.3% of cases using SPINO-based assessment, 19.5% using RECIST1.1, and 6.8% using MDACC. Conclusions: The SPINO recommendations provide a practical and comprehensive framework for radiographic response assessment in monitoring spinal metastases treated with a combination of surgical decompression and adjuvant SSRS. Full article

Background/Objectives: In situ vaccines that directly release endogenous tumor antigens in situ to elicit anti-tumor immune responses without exogenous antigen preparation have emerged as a promising cancer immunotherapy strategy, due to their enhanced safety by local immunization that minimizes systematically adverse reactions. However, the anti-tumor efficacy of most in situ vaccines is affected by their limited access to tumors in distant sites and the toxicity of the adjuvants contained. Methods: To overcome these shortcomings, the present study explored the feasibility of utilizing extracellular vesicles from the probiotic bacteria Lactococcus lactis as both immune activators and drug carriers, which were formulated into nanoparticles to target distant tumors. Results: Using confocal microscopy and flow cytometry, we confirmed that the Lactococcus lactis-derived extracellular vesicles possess adjuvant activity that promoted the maturation of dendritic cells without affecting their viability or apoptosis rate. Moreover, the Lactococcus lactis-derived extracellular vesicles, both alone and when carrying the drug doxorubicin, could target and accumulate in solid tumor tissues via the enhanced permeability and retention effect. Interestingly, compared to healthy cells, the Lactococcus lactis-derived extracellular vesicles tended to be taken up more by tumor cells and readily released their encapsulated doxorubicin in the acidic tumor environment, which resulted in their enhanced reactive oxygen species production and immunogenic cell death. Ultimately, systemic administration of Lactococcus lactis-derived extracellular vesicle-encapsulated doxorubicin greatly increased the anti-tumor efficacy by boosting the number of infiltrating dendritic cells and CD8⁺ T cells in the tumor tissues and doxorubicin-mediated immunogenic cell death. Conclusions: Collectively, this study demonstrated that the probiotic Lactococcus lactis-derived extracellular vesicles are both safer adjuvants and effective drug carriers with immunostimulatory activity and tumor-targeting capability, shedding an interesting light on this vaccine design platform for future cancer immunotherapy. Full article

Background: Papillary thyroid carcinoma (PTC) is typically associated with an excellent prognosis, and distant metastases are uncommon. When present, metastases most frequently involve the lungs and bones, while intra-abdominal dissemination is exceedingly rare and poorly characterized. The optimal management of such atypical metastatic patterns, particularly the role of surgery, remains undefined. Case Presentation: A 69-year-old female presented eight years after total thyroidectomy, cervical lymph node dissection, and radioactive iodine therapy for PTC with progressive abdominal pain, nausea, vomiting, and painful abdominal wall and bilateral inguinal masses. Imaging demonstrated extensive metastatic disease involving the anterior abdominal wall, gastrohepatic ligament, pelvis, retroperitoneum, diaphragm, urinary bladder, uterus, and inguinal lymph nodes, which was confirmed on biopsy as metastatic PTC. Serum thyroglobulin was markedly elevated at 473 µg/L. Given the bulky, symptomatic, and progressive disease refractory to prior radioactive iodine therapy, a multidisciplinary tumor board recommended cytoreductive surgery with palliative intent. The patient underwent resection of eleven major metastatic deposits over five hours without intraoperative complications, achieving complete macroscopic clearance of intra-abdominal disease. Histopathology confirmed metastatic PTC in all specimens. Postoperative serum thyroglobulin declined to 107 µg/L following surgery and adjuvant radioactive iodine therapy, and the patient reported complete resolution of abdominal symptoms at follow-up. Conclusions: This case highlights the potential role of cytoreductive surgery as a palliative strategy in carefully selected patients with advanced intra-abdominal metastatic PTC. Such rare presentations underscore the importance of long-term surveillance and multidisciplinary decision-making. Further case accumulation is needed to better define optimal management strategies and patient selection criteria in this setting. Full article

Mycobacterium abscessus subsp. massiliense (Mabs) is an emerging nontuberculous mycobacterium associated with difficult-to-treat infections due to intrinsic antimicrobial resistance, intracellular persistence, biofilm formation, and limited responsiveness to currently available therapeutic regimens. In this context, adjuvant strategies targeting iron-dependent metabolic pathways and metal homeostasis may enhance the efficacy of conventional antimicrobials. This study investigated deferoxamine (DFO), a clinically approved iron chelator, as a potential adjuvant against Mabs using integrated in vitro and in silico approaches. Cytocompatibility was assessed using an MTT assay in RAW 264.7 macrophages and a hemolysis assay in human erythrocytes. Antimicrobial activity was evaluated through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays, while interactions with clarithromycin (CLA) and amikacin (AMK) were assessed using the checkerboard method. Effects on virulence-associated phenotypes were examined through biofilm formation assays and protein quantification in extracellular vesicle-enriched fractions. Intracellular activity and modulation of inflammatory mediator gene expression were investigated in Mabs-infected RAW 264.7 macrophages through colony-forming unit (CFU) recovery and reverse transcription quantitative polymerase chain reaction (qPCR). DFO exhibited low cytotoxicity and negligible hemolytic activity under the tested conditions. Direct antimicrobial testing revealed a predominantly bacteriostatic profile (MIC = 9.75 µg/mL; MBC > 10 mg/mL), whereas checkerboard analysis suggested a synergistic interaction with CLA (FICI = 0.047), which requires further confirmation by time-kill or CFU-based combination assays. Furthermore, DFO reduced biofilm biomass, decreased protein levels in vesicle-enriched fractions, lowered intracellular bacterial burden, and modulated cytokine gene expression in infected macrophages. Molecular docking, ADME/Tox, and PASS analyses generated exploratory hypotheses regarding potential molecular interactions and pharmacological properties. Overall, these findings support DFO as a promising experimental adjuvant candidate for further investigation against Mabs, particularly in combination with clarithromycin. However, confirmation of a putative iron-restriction-associated mechanism and its translational relevance will require validation in additional clinical isolates, iron-rescue experiments, mature biofilm models, and in vivo studies. Full article

Glioblastoma multiforme (GBM) remains one of the most aggressive primary brain tumors with poor prognosis despite multimodal therapy. Photodynamic therapy (PDT) using indocyanine green (ICG) is an emerging adjuvant approach aimed at eliminating residual tumor cells after resection. While ICG-PDT exerts cytotoxic effects, its impact on molecular pathways regulating programmed cell death in glioma cells is not fully understood. In this study, T98G and U-118MG glioblastoma cells were divided into four groups: untreated control, light-only (10 min broadband irradiation), ICG-only (15 min incubation), and ICG-PDT (15 min ICG + 10 min broadband irradiation). Relative mRNA expression of apoptosis-related genes (BAX, BCL2, CASP3, FAS) and ferroptosis-related genes (GPX4, ACSL4, SLC7A11, GCH1) was quantified 24 h post-treatment by RT-qPCR using the 2^−ΔΔCt method. ICG-PDT significantly reduced cell viability to 67.79% ± 3.39% (vs. 86.66% ± 4.33% in control), confirming effective phototoxicity. No statistically significant differences in mRNA levels were observed for any of the investigated genes across the groups (one-way ANOVA and Kruskal–Wallis, all p > 0.05). The largest non-significant deviation was a mild decrease in GPX4 (fold change 0.87) in the ICG-PDT group. Fluctuations in GCH1 were accompanied by high variance, likely reflecting technical noise rather than a true biological trend. The mRNA BAX/BCL2 ratio remained stable (~30) across all conditions. In contrast, the U-118MG line showed greater transcriptional sensitivity, with statistically significant decreases in CASP3 (p = 0.012) and ACSL4 (p = 0.031) expression, along with downward trends in BCL2 and GPX4 following ICG-PDT. ICG-PDT does not induce significant transcriptional changes in the analyzed genes T98G at the 24 h time point under the applied experimental conditions. In U-118MG cells, moderate transcriptional engagement of both apoptotic and ferroptotic routes was observed. Further studies at the protein and functional levels, across multiple time points and models, are warranted to fully elucidate the mechanisms of ICG-PDT in glioblastoma. Full article

Adjuvant XELOX (capecitabine plus oxaliplatin) and TS-1 (S-1) monotherapy are both guideline-recommended following D2 gastrectomy for gastric cancer, yet head-to-head real-world data exclusively in stage III disease remain scarce. Using Taiwan’s National Health Insurance Research Database linked to the Taiwan Cancer Registry–Long Form, we identified stage III gastric cancer patients who underwent D2 gastrectomy (2010–2019) and received adjuvant XELOX or TS-1 for ≥3 months. Propensity score matching balanced chemotherapy and non-chemotherapy cohorts (1706/group). Overall survival (OS) was the primary endpoint; disease progression was defined as initiation of FOLFOX salvage chemotherapy (used as a pragmatic proxy for disease recurrence). A second propensity score matching was performed directly between XELOX (n = 533) and TS-1 (n = 893) groups, yielding 490 matched pairs with well-balanced baseline characteristics. Multivariable Cox regression was adjusted for sex, age, comorbidities, and Charlson Comorbidity Index. TS-1 was associated with significantly better OS (adjusted HR 0.73, 95% CI 0.61–0.86; p < 0.001) and lower progression (adjusted HR 0.38, 95% CI 0.23–0.62; p < 0.001) versus XELOX; the corresponding 3-year OS was approximately 65.4% for TS-1 versus 56.8% for XELOX, and extrapolated 5-year OS approximately 50.2% versus 41.7%, respectively (note: these 5-year estimates are Kaplan–Meier projections beyond the mean follow-up of ~2.6 years and carry substantial uncertainty; they should be interpreted with caution). Benefits were confined to stage IIIA (OS HR 0.64, 95% CI 0.45–0.89; p = 0.009; interaction p = 0.006; progression HR 0.29, 95% CI 0.11–0.76; p = 0.011), with comparable outcomes in IIIB and IIIC. Adjuvant TS-1 monotherapy was associated with superior OS and lower disease progression versus XELOX in stage III gastric cancer, particularly in stage IIIA; these findings are hypothesis-generating and warrant confirmation in prospective randomized trials, whereas in stage IIIB/IIIC outcomes were comparable between the two regimens. Full article

Background: Colorectal cancer (CRC) remains a leading cause of mortality worldwide, with a significant proportion of patients presenting with metastatic disease (mCRC). While molecularly targeted therapies, including anti-EGFR and anti-VEGF agents, have improved survival outcomes, their efficacy is often limited by drug resistance, toxicity, and high costs. There is a growing need for sustainable strategies to enhance therapeutic efficacy. Methods: This review explores the emerging role of plant-derived compounds as synergistic adjuvants. Specifically, PubMed, Scopus, and Web of Science were searched for English-language articles published between January 2004 and June 2026, using combination of terms related to colorectal cancer, metastatic disease, anti-EGFR/anti-VEGF targeted therapy, phytochemicals/natural products, and gut microbiota; both primary studies and reviews were eligible. Results: Targeted therapies such as cetuximab and bevacizumab are the standard of care but face challenges related to RAS/BRAF mutations and primary tumour location. Clinical data demonstrate that while cetuximab improves overall survival in patients with RAS wild-type, left-sided tumours (median OS 31 vs. 26 months; HR 0.76, p = 0.012), progression-free survival remains comparable to that of bevacizumab. Concurrently, natural products like Vitis vinifera, Dendrobium candidum, and quercetin demonstrate significant preclinical potential in inhibiting angiogenesis, inducing apoptosis, and modulating the tumour microenvironment. The gut microbiome, particularly Fusobacterium nucleatum (whose reported prevalence varies widely across cohorts and reaches up to ~98% of CRC tissues only in selected series), has emerged as a key factor in chemoresistance. It should be emphasised that the great majority of the phytochemical-targeted therapy combinations discussed here are currently supported primarily by preclinical (in vitro and animal) studies rather than by clinical trials. Conclusions: Integrating evidence-based phytochemicals with conventional targeted therapies is a mechanistically compelling and potentially sustainable strategy that may enhance therapeutic efficacy, help overcome resistance, and mitigate adverse effects in mCRC management. However, because current support is largely preclinical, these combinations should be regarded as hypothesis-generating and require validation in prospective, biomarker-stratified clinical trials before clinical adoption. Full article

Background: Predicting long-term outcomes after pulmonary metastasectomy for colorectal cancer remains challenging because existing prognostic methods lack precision. We developed and validated a prognostic scoring system derived from a major international meta-analysis to improve risk stratification and to evaluate the benefit of adjuvant chemotherapy across risk groups. Methods: Using a Japanese registry of 819 patients who underwent lung resection between 2010 and 2019, we constructed a 0–13-point score based on eight variables including tumor size, number, biological markers, and intrathoracic lymph node status, which may require intraoperative or pathological confirmation. Granular data on chemotherapy regimens, timing, and duration were unavailable. Patients were classified as low, intermediate, or high risk. The primary analysis used inverse probability of treatment weighting to adjust for baseline imbalances; however, only 819 of 1657 patients (49.4%) had complete prognostic data, introducing potential selection bias. Results: The score separated patients into three groups with distinct five-year survival rates: 81.1% (low), 67.8% (intermediate), and 59.1% (high). In high-risk patients, chemotherapy was associated with improved overall survival but did not delay recurrence. In low-risk patients, chemotherapy correlated with reduced recurrence-free survival, a finding that persisted after adjustment. Conclusions: This validated scoring system aids individualized surgical decision making by identifying patients unlikely to benefit from routine postoperative chemotherapy. Observed survival advantages in high-risk patients may reflect selection of fitter individuals rather than direct treatment effects, underscoring the need to address selection bias in future trials. Full article

The continuous evolution of the SARS-CoV-2 virus, marked by the emergence of new variants, poses a significant threat to the efficacy of existing vaccines. However, a promising approach to addressing vaccine failure caused by viral mutations (particularly in the spike protein) is the development of a variant-proof (conserved), non-spike, multiepitope universal nanostructure vaccine with multifunctionality, biocompatibility, self-adjuvanticity, and structural similarity to pathogens in terms of size and shape. This study aimed to design a self-assembled nanostructure vaccine (SANV) featuring pentameric and trimeric coiled-coil peptide motifs, as well as other functional motifs, including epitopes, TAT, PADRE, and adjuvant. The cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and B lymphocyte (BL) epitopes of SANV were screened from the IEDB with more than 50% individual predicted population coverage (PPC) and fused using linkers to enable self-assembly. The multimerization of the 24 SANV monomers was modeled using the GalaxyHomomer and AlphaFold web servers. Subsequently, the leading SANV constructs with (SANVa9) and without (SANVb6) adjuvant were analyzed for their physicochemical profiles and assessed for antigenicity, allergenicity, solubility, and antioxidant potential. Furthermore, the molecular interactions, specificity, and stability of SANVa9 and SANVb6 with the broadly neutralizing sarbecovirus antibody 5817 and toll-like receptors (TLR2, TLR3, and TLR7) were analyzed using molecular docking and simulation over a 100-nanosecond time scale. Finally, the comparative immune simulation profiles of SANVa9 and SANVb6 with controls indicated stronger, broad-spectrum immune responses that could be translated into in vitro and in vivo studies and warrant further evaluation before clinical use. Full article

Simple and non-invasive transdermal vaccination is an attractive alternative to conventional injection-based immunization. However, the effectiveness of transdermal vaccines is often constrained by the stratum corneum barrier. Although the use of solid-in-oil (S/O) nanodispersion technology has successfully facilitated skin permeation to induce an immunological response, the antibody titers remain suboptimal. Herein, a dectin-1 selective ligand, laminarin, was used as an immunostimulatory adjuvant to enhance the immune response. S/O nanodispersions loaded with laminarin and ovalbumin (OVA) were systematically developed and characterized in terms of particle size, in vitro OVA release behavior, and skin permeation performance using excised mouse skin. In vivo immunization via transcutaneous administration was performed to evaluate biocompatibility and antigen-specific immunoglobulin-G (IgG) responses. Laminarin-loaded S/O nanodispersions demonstrated long-term stability and efficient ex vivo skin permeability. All the prepared laminarin-loaded S/O nanodispersions showed increased OVA-specific IgG responses compared with the laminarin-free S/O formulation. Among the formulations, the S/O nanodispersion containing OVA and laminarin at a 1:4 weight ratio induced 20-fold higher OVA-specific IgG responses than PBS and 7-fold higher responses than laminarin-free S/O formulations. This study clearly demonstrates the potential of laminarin-loaded S/O nanodispersions as a non-invasive vaccine delivery platform for enhancing antigen-specific antibody responses. Full article

Background and Clinical Significance: Malignant transformation of giant cell tumor of bone (GCTB) is a rare but clinically significant event, most commonly associated with radiotherapy, denosumab therapy, or recurrent disease. Secondary malignant transformation occurring in the absence of recognized risk factors is exceptionally uncommon. We report a rare case of high-grade sarcomatous transformation of proximal humeral GCTB after a prolonged latency period without prior radiotherapy, denosumab exposure, or documented recurrence; Case Presentation: A 27-year-old female initially presented with right shoulder pain and was diagnosed with proximal humeral GCTB. She underwent intralesional curettage and bone grafting, with histopathological confirmation of benign GCTB. Nine years later, she developed progressive shoulder pain, functional limitation, and systemic symptoms. Imaging demonstrated an aggressive lytic lesion with cortical destruction and soft-tissue extension involving the proximal humerus. Repeat curettage and histopathological evaluation revealed high-grade spindle cell sarcoma consistent with malignant transformation of GCTB. The patient received neoadjuvant chemotherapy followed by wide resection and endoprosthetic reconstruction of the proximal humerus, with additional adjuvant chemotherapy postoperatively. At two-year follow-up, she remained disease-free with excellent functional recovery and satisfactory quality of life; Conclusions: This case highlights the potential for delayed malignant transformation of GCTB even in the absence of established predisposing factors. Clinicians should maintain long-term vigilance in patients treated for GCTB, particularly when new pain, functional decline, or aggressive radiologic features develop years after initial treatment. Early recognition and multidisciplinary management are essential to optimize oncologic and functional outcomes. Full article

Background: Lipopolysaccharide (LPS) functions as a Toll-like receptor 4 (TLR4) agonist that triggers innate immunity; however, structural variations between pathogenic and commensal bacteria distinctly influence its immunostimulatory profile. This study evaluated the immunostimulatory activity of LPS derived from the commensal bacterium Hafnia alvei and explored its potential as an exploratory vaccine adjuvant. Methods: Cytokine induction was evaluated in immune cells across diverse host species, and receptor activation was assessed via reporter assays. To investigate in vivo immunogenicity and preliminary tolerability, H. alvei LPS was formulated into a prototype oil-in-water (O/W) emulsion utilizing ovalbumin (OVA) as a model antigen. Results: LPS from H. alvei strain BA2000346 exhibited immunostimulatory activity comparable to that of Escherichia coli, while inducing greater TNF-α expression than pathogenic Salmonella and Pseudomonas strains. Distinct from E. coli LPS, it demonstrated the capacity to activate both TLR4 and the mannose-recognizing Dectin-2 receptor in reporter systems. This cytokine induction was consistent across various strains and host species. Furthermore, the prototype O/W emulsion formulation enhanced antigen-specific humoral and cellular immune responses while demonstrating preliminary tolerability based on body-weight monitoring and visual clinical observation. Conclusions: H. alvei-derived LPS exhibits TLR4 and Dectin-2 agonist activity in vitro. When synergized with an O/W emulsion delivery system, it provides a preliminary indication of cross-species stimulatory potential and supports further investigation as a hypothesis-generating platform for future vaccine adjuvant development. Full article

Fasciolosis, caused by the liver fluke Fasciola gigantica, remains a major parasitic disease affecting livestock in tropical regions and results in substantial economic losses. Although anthelmintic drugs are widely used for disease control, increasing reports of drug resistance highlight the need for alternative strategies such as vaccination. In this study, a recombinant digestive protein-based chimeric antigen (rFgCHI-DP) composed of three F. gigantica antigens—cathepsin L1 (FgCL1), cathepsin B1 (FgCB1), and saposin-like protein 2 (FgSAP2)—was designed and expressed in Escherichia coli. The mature regions of these proteins were sequentially linked to form a single chimeric construct. The recombinant protein was successfully expressed and purified under denaturing conditions, producing a protein of approximately 62 kDa. To evaluate its immunogenicity, BALB/c mice were immunized with rFgCHI-DP formulated with Quil A adjuvant. Indirect ELISA revealed that immunization induced antigen-specific IgG responses. Antibody responses showed strong reactivity toward FgCL1 and FgCB1, whereas the response against FgSAP2 was comparatively lower. Western blot analysis further demonstrated that antibodies generated against rFgCHI-DP recognized native parasite antigens. Immunolocalization also revealed that the anti-rFgCHI-DP antibodies could detect targeted antigens in the cecal epithelium of the parasite. These findings indicate that the adult-stage chimeric protein rFgCHI-DP is immunogenic in mice and capable of inducing specific antibody responses against F. gigantica. The results support the potential of rFgCHI-DP as a candidate antigen for future fasciolosis vaccine development. Full article