Search Results (2,327)

This study investigated colostrum source impact on passive immunity transfer and humoral immune development in Polypay lambs. Newborn lambs (4.80 ± 0.33 kg BW) were assigned to four groups: fresh ewe colostrum (FrC; n = 10), frozen ewe colostrum (FZ; n = 11), frozen cattle colostrum (CC; n = 11), or artificial cattle colostrum (AC; n = 11). Lambs received 65 mL/kg colostrum within 4 h post-parturition and were raised on milk replacer, creep feed, and hay until weaned (28 days of age [d]). Immunoglobulin G concentrations were measured at birth and then weekly until 28 d. Lambs were immunized with 1 mL ovalbumin (2 mg/mL PBS) at 35 and 63 d. Ovalbumin-specific antibodies were quantified. A tendency (p = 0.06) suggested FrC yielded higher relative IgG at later time points (7, 14, 21, and 28 d) compared to FZ. Frozen cattle colostrum resulted in significantly (p = 0.02) higher IgG concentrations than AC at 24 h, 7, 14, and 21 d. Fresh ewe colostrum and AC showed a faster (1 week-post-immunization) and more robust (>175%; p ≤ 0.07) ovalbumin humoral response compared to CC lambs 2 to 8 weeks-post-immunization. Fresh ewe colostrum seems to provide the best passive and adaptive immunity compared to other colostrum sources. Moreover, our results suggest that the best colostrum alternative for newborn lambs is frozen ewe colostrum. Full article

Human T-lymphotropic virus type 1 (HTLV-1) infection is associated with a spectrum of clinical outcomes, ranging from lifelong asymptomatic carriage to severe conditions such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T-cell leukemia/lymphoma (ATLL). Although antibody responses are known to shape immune regulation, the functional relevance of IgG idiotype repertoires in HTLV-1 pathogenesis remains poorly understood. This study investigated the immunomodulatory effects of IgG from individuals with distinct HTLV-1 clinical outcomes. IgG was purified from pooled serum samples of asymptomatic carriers (ACs), HAM/TSP, and ATLL patients and used to stimulate peripheral blood mononuclear cells (PBMCs) from healthy donors. Cytokine production in CD4⁺, CD8⁺, and γδ T cells was assessed by flow cytometry. Additionally, proteome-wide IgG reactivity was evaluated using a human protein microarray encompassing over 21,000 proteins, and bioinformatic analyses were conducted to identify protein–protein interaction networks and tissue-specific autoreactivity. HAM/TSP-derived IgG selectively enhanced IFN-γ production in all T-cell subsets and suppressed IL-4 in CD4⁺ T cells. ATLL-derived IgG induced IL-9 and IL-13 production in CD4⁺ T cells, and both HAM/TSP and ATLL IgG elevated IL-13 levels in CD8⁺ T cells. Microarray data revealed distinct autoreactive IgG profiles across clinical groups, targeting immune-related proteins, apoptotic regulators, and proteins expressed in T cells, monocytes, and non-immune tissues such as brain and testis. Notably, no functional or structural clustering was observed in protein–protein interaction networks, suggesting these reactivities reflect complex, idiotype-specific immune alterations rather than compensatory responses. The present findings suggest that HTLV-1 infection may be associated with the development of distinct IgG repertoires that potentially modulate cytokine responses and exhibit broad reactivity toward human proteins. Such patterns could contribute to immune dysregulation and may partially explain the divergent clinical trajectories observed in HAM/TSP and ATLL. Further investigations are warranted to validate these observations at the individual level and to clarify their mechanistic relevance in disease progression. Full article

Background/Objectives: Subunit vaccines composed of purified proteins and adjuvants offer excellent safety, but often generate short-lived immunity due to rapid antigen clearance and limited antigen-presenting cell engagement. Sustained, localized delivery of antigen and adjuvant may improve the magnitude and durability of the immune response without compromising safety. This study evaluated an in-situ polymerizing type I oligomeric collagen (Oligomer) scaffold to localize antigen/adjuvant at the injection site and prolong antigen presentation. Methods: Mice were immunized intramuscularly with ovalbumin (OVA) and CpG oligonucleotide adjuvant delivered alone or co-formulated with Oligomer. Antibody response and inflammation at the injection site were assessed post-booster at early (Day 32) and late (Day 68) time points. Antigen retention and dendritic cell trafficking to draining lymph nodes were evaluated using fluorescently labeled OVA. Results: The Oligomer scaffold retained vaccine antigen at the injection site without eliciting a material-mediated foreign body response. Co-delivery of OVA and CpG within the scaffold enhanced germinal center activity, increased follicular helper T cells and germinal center B cells, and skewed CD4⁺ T cells toward a Th1 phenotype. Humoral responses were greater and more durable, with higher OVA-specific IgG, IgG1, and IgG2a titers and an increased number of bone marrow antibody-secreting cells persisting through Day 68. Antigen-positive dendritic cells, including both resident and migratory subsets, were elevated in draining lymph nodes, indicating enhanced antigen transport. No anti-mouse collagen I antibodies were detected, confirming the maintenance of collagen self-tolerance. Conclusions: The Oligomer delivery platform functioned as a localized, immunotolerant vaccine depot, sustaining antigen availability and immune cell engagement. This spatiotemporal control enhanced germinal center responses and generated a more robust, durable humoral immune response, supporting its potential to improve subunit vaccine efficacy while maintaining an excellent safety profile. Full article

Influenza is a highly contagious disease and is transmitted by the upper respiratory tract. Vaccination is an effective strategy to prevent and control seasonal influenza. The current predominant split-inactivated influenza vaccine presents a high safety profile but has weak immunogenicity. The addition of adjuvants is one method to optimize the immunogenicity of the seasonal influenza vaccine. In this study, we compared the effect of aluminum (Alum), MF59-like adjuvant AddaVax, and ISA71 VG adjuvants for the seasonal split influenza vaccine in a mouse model based on the induction of influenza-virus-specific antibody levels, body weight changes, and survival rates after lethal challenge. Two very low and sub-optimal HA doses, 0.003 µg and 0.01 µg, representing the calculated amount of HA from the A/California/07/2009 (H1N1) strain only per mouse dose, were selected and used in this study. The 0.003 µg antigen (Ag) plus AddaVax showed the best adjuvant effect among these three adjuvants. The 0.01 µg Ag plus ISA 71 VG induced the highest total IgG, IgG1, and IgG2a. Both the 0.003 µg and 0.01 µg Ag plus AddaVax protected all the immunized mice from the lethal challenge, and Alum exhibited the protective potency intermediate between that of the AddaVax and ISA 71VG. The 0.01 µg Ag plus one of these three adjuvants could enhance the efficacy of the split influenza vaccine against lethal challenge. Therefore, AddaVax is the first candidate for the further development of the adjuvanted split seasonal influenza vaccine among these three adjuvants. These initial findings offer valuable guidance for selecting promising adjuvanted influenza vaccine formulations. Full article

Over the past few decades, Fc fragment-conjugated proteins, such as Protein A, have been extensively utilized across a range of applications, including antibody purification, site-specific immobilization of antibodies, and the development of biosensing platforms. In this study, building upon our group prior research, we designed and screened an affinity DNA functional ligand (A-DNAFL) and experimentally validated its binding affinity (K_D = 6.59 × 10⁻⁸) toward mouse IgG antibodies, whose binding performance was comparable to that of protein A. Systematic evaluations were performed to assess the binding efficiency under varying pH levels and ionic strength conditions. Optimal antibody immobilization was achieved in PBST-B buffer under physiological pH 7.2–7.4 and containing approximately 154 mM Na⁺ and 4 mM K⁺. Two competitive binding assays confirmed that the A-DNAFL binds to the Fc fragment of murine IgG antibody. Furthermore, molecular docking simulations were employed to investigate the interaction mode, revealing key residues involved in binding as well as the contributions of hydrogen bonding and hydrophobic interactions to complex stabilization. Leveraging these insights, A-DNAFL was utilized as a tool for oriented immobilization of antibodies on the sensing interface, enabling the construction of an immunosensor for ricin detection. Following optimization of immobilization parameters, the biosensor exhibited a detection limit of 30.5 ng/mL with the linear regression equation is lg(Response) = 0.329 lg(C_ricin) − 2.027 (N = 9, R = 0.938, p < 0.001)—representing a 64-fold improvement compared to conventional protein A-based methods. The system demonstrated robust resistance to nonspecific interference. Sensing interface reusability was also evaluated, showing only 8.55% signal reduction after two regeneration cycles, indicating that glycine effectively elutes bound antibodies while preserving sensor activity. In summary, the A-DNAFL presented in this study represents a novel antibody-directed immobilization material that serves as a promising alternative to protein A. It offers several advantages, including high modifiability, low production cost, and a relatively small molecular weight. These features collectively contribute to its broad application potential in biosensing, antibody purification, and other areas of life science research. Full article

Background and Aims: C1q is an autoantigen in different autoimmune disorders, Systemic Lupus Erythematosus (SLE) and Lupus Nephritis (LN) among them. The two functional domains of C1q, the collagen-like region (CLR) and the globular head region (gC1q), are frequently recognized by autoantibodies in SLE and LN when C1q is immobilized. We studied whether autoantibodies to C1q in SLE and LN patients recognized C1q as a soluble autoantigen and whether the act of immobilization was a prerequisite for the recognition of C1q autoepitopes localized on gC1q domains. Methods: The interaction of soluble C1q and its globular fragments ghA, ghB, and ghC with immobilized IgG autoantibodies (and vice versa) from sera of 48 patients with SLE and LN was studied with ELISA. Data were compared using Spearman correlation coefficient. Fluorescence spectroscopy was used to study the interaction between C1q and LN IgG autoantibodies both presented in solution. Results: We found that anti-C1q autoantibodies from SLE and LN patients specifically bound C1q and gC1q fragments, ghA, ghB, and ghC, both as immobilized and soluble antigens. Correlation analysis indicated a negative correlation between the levels of autoantibodies against immobilized and soluble C1q and immobilized and soluble gC1q fragments which indicates different epitopes when these proteins were recognized as autoantigens in soluble and immobilized conformations. Conclusions: Serum C1q in patients with SLE is a target molecule for binding from anti-C1q autoantibodies. The gC1q region undergoes a conformational change in an immobilized and a soluble form, thus exposing different epitope-binding sites. Full article

Background: Latvia is one of the most endemic tick-borne encephalitis (TBE) countries in Europe. However, assessing TBE seroprevalence and true infection rates has been challenging. Current diagnostics cannot reliably distinguish between immune responses induced by natural infection from those induced by vaccination, especially in TBE endemic countries with recommended immunisation programmes. A recently developed ELISA targeting antibodies against TBEV non-structural protein 1 (NS1) offers improved specificity for natural infection and can differentiate among three clinically relevant TBEV subtypes. Methods: We conducted a cross-sectional TBEV seroprevalence study in the Latvian population during 2019–2022, consisting of two pools: 1020 residents living in different regions of Latvia and 200 random Latvian Biobank blood samples. We used the standard anti-TBEV IgG ELISA (VIDITEST, VIDIA, Czech Republic) for the screening and detection of TBEV (whole virus) IgG antibodies and the newly established research-use anti-TBEV NS1 IgG ELISA for the detection of subtype-specific TBEV NS1 IgG antibodies against three virus subtypes: European, Siberian and Far Eastern. Results: The total TBEV seroprevalence among 1020 residents was 39.7%, representing all age cohorts of the population from all regions of Latvia. In total, 33.4% of the enrolled population were vaccinated against TBE with at least one dose of the TBE vaccine. Among the unvaccinated population, 16.3% had positive TBEV-specific IgG antibodies by standard ELISA. On the contrary, NS1-specific antibodies, indicating past natural infection, were detected in only 4.3% of the overall study population. Subtype differentiation revealed infections from all three major TBEV subtypes present in Latvia. Conclusions: In conclusion, this population-based study highlights a high risk of TBE in Latvia, with substantial seroprevalence even among unvaccinated individuals. The NS1-based ELISA enhances the accuracy of TBE surveillance and offers important clinical utility by facilitating more reliable diagnosis and case classification, regardless of vaccination status. Full article

Background and Objectives: Primary Epstein–Barr virus (EBV) infection in pediatric kidney transplant recipients with donor/recipient mismatch (D+/R−) carries the highest risk of post-transplant lymphoproliferative disorder (PTLD). Current prophylactic strategies are not standardized. Intravenous immunoglobulins (IVIG), containing anti-EBV antibodies, have been proposed as a potential preventive option, but evidence is lacking. This single-center retrospective case–control study evaluated the efficacy of serial IVIG administration in preventing primary EBV infection and promoting long-term immunity in this high-risk population. Materials and Methods: We retrospectively analyzed 26 pediatric kidney transplant recipients (age 1–18 years) with EBV D+/R− mismatch and a median follow-up of 7.5 years. Fourteen patients received scheduled IVIG infusions (200 mg/kg monthly for six months post-transplantation), while twelve received no EBV-directed prophylaxis. The primary endpoint was the cumulative incidence of primary EBV infection, defined as EBV-DNA > 1000 copies/mL in peripheral blood. The secondary endpoint was Epstein–Barr Nuclear Antigen-Immunoglobulin G (EBNA-IgG) seroconversion. Results: Patients receiving IVIG were significantly younger than controls (median age 4.2 vs. 10.8 years, p = 0.01). No significant variations were observed between groups in renal function or immunosuppressive levels during follow-up. IVIG prophylaxis was unexpectedly linked to a higher cumulative incidence of EBV infection compared with controls (64% vs. 25%, p = 0.047). Time-to-event analysis confirmed an increased, although not statistically significant, risk of EBV acquisition in the IVIG group (Hazard Ratio [HR] 3.24, 95% Confidence Interval [CI] 0.87–12.01; p = 0.079). EBV-specific immunity, assessed by EBNA-IgG seroconversion, was comparable between groups (HR 1.78; p = 0.45), confirming no immunological advantage of IVIG. One IVIG-treated patient (7.1%) developed PTLD, while none did in the control group. Conclusions: Scheduled IVIG administration during the first six months after transplantation does not constitute an effective strategy to prevent primary EBV infection or to enhance long-term immunity in high-risk EBV D+/R− pediatric kidney recipients and may even increase susceptibility to viral acquisition. These findings argue against the use of IVIG as EBV prophylaxis in this population. Full article

This study aimed to characterize Mycoplasmosis bovis strain 16M—a highly virulent isolate from one Chinese outbreak—as a candidate for challenge models and inactivated vaccine development. We assessed strain 16M through morphological observation, PCR identification, drug susceptibility testing, growth titer and biofilm quantification, immunological profiling, and calf challenge experiments. We used genomic resequencing to evaluate the genetic stability across 150 passages. Classified as the prevalent ST52 lineage in China, strain 16M showed phylogenetic proximity to strain 08M and exhibited multidrug resistance (notably to macrolides). It achieved higher titers and stronger biofilm formation than other isolates and the reference strain PG45. In calves, intratracheal inoculation with 16M induced universal infection, severe pulmonary consolidation, and peribronchial cuffing, with significantly higher disease scores (p < 0.01). The inactivated 16M vaccine elicited elevated antigen-specific IgG titers, PBMC proliferation, and IFN-γ production versus PG45. Post challenge, immunized calves showed reduced pathological lesions, shorter bacterial shedding, and lower disease scores than the infected controls (p < 0.05). Genetic stability was confirmed for virulence-associated genes (e.g., adhesion proteins), with stable titers and biofilm production within 50 generations. Strain 16M combines high virulence for challenge modeling and industrial-scale vaccine suitability, owing to its robust growth, stable immunogenicity, and genetic consistency. Full article

Background: While host immune responses to SARS-CoV-2 vaccination are routinely assessed through IgG measurements, less is known about the temporal dynamics of vaccine-induced cellular immunity, particularly in individuals who fail to develop detectable IgG antibodies after COVID-19 vaccination. Objective: To investigate the development and timing of T-cell immunity following SARS-CoV-2 vaccination in antibody-non-responders to COVID-19 vaccination. Methods: A cross-sectional analysis was conducted on COVID-19-naive individuals who had received full SARS-CoV-2 vaccination, categorized by SARS-CoV-2 IgG serostatus. T-cell response was evaluated using the IGRA methodology T-SPOT^®.COVID (Oxford Immunotec, Abingdon, Oxfordshire, UK). T-cell response rates and levels were compared between SARS-CoV-2 seropositive and seronegative groups, and a temporal cutoff analysis was applied to examine trends in T-cell response over time. Results: Within the seronegative group, IgG levels showed a strong negative correlation with time since vaccination (Spearman ρ = −0.65, p < 0.001), while T-cell response levels exhibited a weak positive time-dependent trend (ρ = 0.15, p = 0.019). Temporal cutoff analysis identified a critical time-point beginning at 80 days post-vaccination, after which both T-cell response rates and levels were significantly higher. Specifically, individuals tested after 80 days showed increased median T-cell response levels (U = 4205, p < 0.001) and higher positive T-cell response rate (67% vs. 38%, Χ² = 17.06, p < 0.001). Conclusions: Cellular immunity response against SARS-CoV-2 may emerge later than expected in antibody-non-responders to COVID-19 vaccination, with the 80-day post-vaccination mark emerging as a critical time point. Our results support the inclusion of cellular assays in post-vaccination monitoring and emphasize the need to reconsider the timing and criteria for evaluating vaccine response. Full article

Background/Objectives: Varicellovirus bovinealpha1 and Varicellovirus bovinealpha5 (BoAHV-1 and BoAHV-5), respectively, are widely distributed pathogens that cause distinct clinical conditions in cattle including infectious bovine rhinotracheitis, infectious pustular vulvovaginitis/balanoposthitis, and meningoencephalitis. Due to the establishment of viral latency, controlling these infections is challenging, and vaccination remains the most effective strategy. In this study, vaccine candidates targeting both BoAHV-1 and BoAHV-5 were developed. Methods: A synthetic gene encoding immunodominant epitopes from the gB and gD proteins and tegument phosphoprotein of BoAHV-1 and BoAHV-5 was designed to produce a multi-epitope recombinant antigen, expressed both in a prokaryotic system (RecBoAHV) and by a modified vaccinia Ankara (MVA-BoAHV) viral vector. The binding affinity of MHC-I to bovine leukocyte antigens (BoLA) was predicted using the NetMHCpan tool (version 4.1). The immunogenicity of the vaccine candidates was evaluated in rabbit and mouse models, using prime-boost immunization protocols. Sera from bovines naturally infected with BoAHV-1 and/or BoAHV-5 were used to evaluate the chimeric protein antigenicity. Immune responses were assessed by indirect ELISA and Western blot. Results: The recombinant multi-epitope protein was effectively recognized by IgG and IgM antibodies in sera from cattle naturally infected with BoAHV-1 or BoAHV-5, confirming the antigenic specificity. Both RecBoAHV and MVA-RecBoAHV induced strong and specific humoral immune responses in rabbits following a homologous prime-boost regimen. In mice, both homologous and heterologous prime-boost protocols revealed robust immunogenicity, particularly after the second booster dose. Conclusions: These findings highlight the immunogenic potential of the RecBoAHV multi-epitope vaccine candidates for controlling BoAHV-1 and BoAHV-5 infections. Further characterization of these vaccine formulations is currently underway in bovine, the target specie. Full article

(1) Background: This study investigated the presence of human papillomavirus (HPV), HPV genotypes, Epstein–Barr virus (EBV), cytomegalovirus (CMV) and herpes simplex virus (HSV) in patients with Head and Neck Cancer (HNC) at both molecular and serological levels. (2) Methods Fifty patients with histopathologically confirmed HNC who were admitted to the Department of Otorhinolaryngology, Istanbul Faculty of Medicine. Viral DNA was detected using quantitative real-time PCR, and serological IgM and IgG antibodies were analyzed using the CMIA method; (3) Results: In blood samples, CMV and HSV DNA were not detected, whereas EBV DNA was identified in 2% and HPV DNA in 4% of patients. In tumor tissues, CMV DNA was detected in 8%, EBV DNA in 10%, and HPV DNA in 6%; HSV DNA is 6%. HPV genotypes 18, 45, and 69 were found in tissue samples. Serologically, IgG positivity for CMV, EBV, and HSV-1 exceeded 90%, whereas IgM positivity was low and not statistically significant; (4) Conclusions: HPV, EBV, and CMV DNA were detected at low frequencies in patients with HNC, while HSV DNA was absent. These findings underline the need for larger multi-center studies and support the consideration of routine viral screening, particularly for HPV, in specific tumor subtypes. Full article

Background: The AFFINITY trial (NCT05890872) is a prospective, non-randomized, open-label, single-arm study evaluating the safety, immunological impact, and preliminary efficacy of Aliya pulsed electric field ablation in patients with solid tumors. Thirty-one patients were enrolled; thirty received lung lesion ablation prior to continuation on standard-of-care treatment. This manuscript reports six-month local control outcomes and immunological response characteristics. Radiological outcomes were assessed using a modified RECIST 1.1, and immunological impact was evaluated via changes in peripheral blood immunocyte populations and detection of immunoglobulins (Ig) to tumor-associated antigens in serum post-ablation. Methods: Twenty-eight patients underwent radiological assessment of ablated lesions at approximately 1-, 3-, and 6-month post-ablation to evaluate local control. Peripheral blood was collected for immune monitoring using flow cytometry and to detect IgG responses to biopsy-specific and tumor-associated antigens. Results: At 6 months, two cohorts emerged: 12 received ablation only, and 16 received ablation plus systemic and/or focal therapies (radiotherapy or second ablation). In the ablation-only group, imaging showed local control in all ablated lesions (8/12 SD, 4/12 PR), suggesting local efficacy without systemic therapy in those patients. Immunophenotyping showed dynamic changes in circulating immune cells, including T and B cell activation. A subset also exhibited modulation of tumor antigen-specific IgG, indicating a systemic humoral response. Conclusions: This analysis provides preliminary evidence that this form of ablation may promote local tumor control and modulate systemic immune function. These findings support the immunogenic potential of this specialized energy and warrant further investigation. Extended 12-month data for the full cohort will be reported in a future manuscript. Full article

Background/Objectives: Allergen-specific immunotherapy remains the only disease-modifying treatment for allergic diseases, and the use of recombinant hypoallergenic derivatives is a promising therapeutic approach. Among these, BTH2 has previously shown efficacy in an acute murine model of allergy induced by Blomia tropicalis. The present study aimed to evaluate both the efficacy and safety of BTH2 in a chronic asthma model induced by B. tropicalis. Methods: A/J male mice (n = 6) were sensitized and chronically challenged with B. tropicalis extract over four months. One group repeatedly received subcutaneous doses of BTH2 (25 µg) for three months (65 doses). Parameters of allergic airway inflammation, antibody profiles, cytokine levels, and markers of AIT success were evaluated in bronchoalveolar lavage fluid, lung tissue, serum, and splenocyte cultures. Results: Repeated BTH2 administration was well tolerated, with no signs of systemic toxicity. BTH2 significantly reduced neutrophilic and eosinophilic airway inflammation, while increasing lymphocytes and regulatory cytokines in the lungs. It suppressed IgE against B. tropicalis allergens, while inducing mucosal IgA responses and systemic IgG, which may be linked to the observed blocking antibody activity in BTH2-treated mice. The treatment also led to downregulation of Th2 cytokines and enhanced expression of regulatory and Th1-associated cytokines, especially IL-10, TGF-β and IFN-γ. Correlation matrix analyses indicated that regulatory cytokines were correlated with beneficial antibody responses and reduced inflammation. Conclusions: BTH2 shows strong therapeutic and immunomodulatory effects in a chronic asthma model induced by B. tropicalis, with a favorable safety profile. These findings support its potential for future clinical trials, including those involving patients with allergic asthma. Full article

Background: In response to the emergence of immune-evasive variants of SARS-CoV-2, this study explores a novel heterologous vaccination strategy using a microparticulate formulation approach that is delivered via oral dissolving film (ODF) formulations into the buccal cavity. Heterologous administration has the potential to generate cross-reactive antibodies, which can be especially beneficial against viruses with ever-mutating variants. Moreover, the microparticulate oral dissolving film-based vaccine approach is a non-invasive vaccine delivery platform. Methods: The vaccine design incorporated whole inactivated Delta and Omicron variants of the virus, administered at prime and booster doses, respectively, effectively encapsulated in a Poly(lactic-co-glycolic) acid (PLGA) polymer matrix, and adjuvanted with Alum to enhance immune activation. Following vaccination, serum, mucosal, and tissue samples were analyzed to evaluate humoral and cellular immune responses against the model antigen, as well as other variants such as Alpha and Beta variants, to understand the cross-reactive response. Result: In vitro evaluations confirmed the vaccine’s safety and its ability to stimulate immune responses. On administering microparticulate oral dissolving films to mice, whole inactivated delta and omicron variant-specific antibodies were observed in serum samples along with neutralizing titers in terminal week. The formulated vaccine showed significant secretory IgA antibody levels in mucosal samples. Moreover, CD4⁺ and CD8a cellular responses were observed in tissue samples of spleen and lymph nodes, along with antibodies (IgG, IgA, and IgM) detected in lung supernatant samples. Humoral and cellular cross-reactive antibodies were observed in the samples. Conclusions: This approach offers a promising platform for developing next-generation vaccines capable of inducing broad immunity. Full article