Search Results (51)

Two endogenous peptides, β-alanyl-L-histidine, named carnosine (Car), and glycyl-L-histidyl-L-lysine (GHK), derived from the matricellular protein Secreted Protein Acidic and Rich in Cysteine (SPARC), share many beneficial functions. The hydrolytic enzyme carnosinase for Car and the low stability for GHK can put into question their antioxidant, antiaggregating, and anti-inflammatory properties. The glycoconjugates of Car with a di- (trehalose, Tre) or polysaccharide (hyaluronan, HA) inhibit carnosinase, while the synthesis of HAGHK derivatives increases the tripeptide stability and protects/delays the biopolymer degradation. A synergic effect between the two components of the glycoconjugates is evident in their consequently preserved protective features. TreCar, HACar, and HAGHK maintain the copper-binding ability of the peptides alone, and the saccharides potentiate the Cu,Zn-superoxide dismutase-like ability of the copper(II) complexes with the glycoconjugates. These peptide derivatives behave as copper ionophores, utilizing Cu²⁺ present in the culture medium; also, an increase in the metal intracellular level occurs with a consequent stimulation of the copper-driven signaling pathways that produce the expression/release of trophic (Brain-Derived Neurotrophic Factor, BDNF, and Bone Morphogenetic Protein 2, BMP-2) and angiogenic (Vascular Endothelial Growth Factor, VEGF) proteins. Copper chaperons for SOD1, CCS, and Antioxidant 1 (Atox-1) are the copper chaperones that act as transcription factors. Full article

Background/Objectives: Cuproptosis, a newly discovered form of programmed cell death, is dependent on the regulation of copper ions. The roles and mechanisms of cuproptosis-related genes (CRGs) in the instability of atherosclerotic plaques are still unclear. Methods: GEO microarray datasets were downloaded to analyze stable and unstable human carotid artery plaques. Differential expression analysis was performed to screen for CRGs from the Molecular Signatures Database (MSigDB). Machine learning was applied to identify key genes and cluster unstable plaque genes. The identified genes were verified by immunohistochemistry (IHC) of human carotid plaque samples, and the effect of ATOX1 on cuproptosis was detected in human umbilical vein endothelial cells (HUVEC). Results: This study identified 27 CRGs differentially expressed between stable and unstable plaques. Five characteristic genes (LC3A, ATP7B, ATOX1, CTR1, and NLRP3) were selected by machine learning. A diagnostic model for unstable plaques was developed based on these genes. The expression of ATOX1 and NLRP3 was increased, while LC3A and ATP7B were decreased in unstable plaques. However, there was no significant change in CTR1. The Cell Counting Kit-8 (CCK-8) assay indicated that inhibiting ATOX1 reduced CuSO₄-induced HUVEC death. Conclusions: CRGs appear to influence atherosclerotic plaque formation. Five key genes (LC3A, ATP7B, ATOX1, CTR1, NLRP3) were identified as being differentially expressed in unstable plaques. Cluster analysis uncovered two subtypes (C1, C2) linked to cuproptosis and immune infiltration in unstable plaques. These genes likely affect atherosclerosis progression by influencing immune cell infiltration, thus impacting plaque stability. Furthermore, the cuproptosis-related gene ATOX1 can regulate CuSO₄-induced HUVEC death. This study contributes to predicting plaque instability and offers potential diagnostic and therapeutic targets. Full article

Background: Lung adenocarcinoma (LUAD), the most prevalent and malignant form of lung cancer subtypes, is in urgent need of additional therapeutic targets and prognostic indicators. Antioxidant 1 (ATOX1) copper chaperone and RhoA/Rho kinase 1 (ROCK1) are novel anti-tumour targets in cancers. However, their prognostic value and synergistic inhibitory effect remain unclear in LUAD. Methods: We re-analyzed the open-access proteomic landscape study of LUAD in 2019 and investigated the prognostic value of ATOX1/ROCK1 expression patterns. Then we verified it immunohistochemically using an independent cohort from our hospital enrolling 35 patients with TNM stage III/IV LUAD. In vitro, double fluorescence was used to confirm the co-expression and location of ATOX1/ROCK1. The CCK—8 assay and Transwell assay were carried out to assess the changes in proliferation and migration of Lewis lung carcinoma (LLC) cells following treatment with ATOX1/ROCK1 si-RNA or inhibitory drugs. Western blot was used to confirm protein expression after si-RNA transfection. Moreover, ATOX1/ROCK1-targeted drugs’ therapeutic effects were further investigated in the LLC allogeneic transplantation model and MNU-induced tumour model. Results: Firstly, according to the ATOX1/ROCK1 expression pattern derived from proteomic data, double-low expression of ATOX1/ROCK1 indicated a better Disease Free Survival (DFS) (log-rank test p = 0.01) and Overall Survival (OS) (log-rank test p = 8.2 × 10⁻³), whose expression was also correlated with the lower expression of MCM family proteins. Further, we verified this prognostic correlation in our cohort. The IHC-defined ATOX1/ROCK1 low subtype also had the best OS (log-rank test p = 2.4 × 10⁻³). In vitro, double fluorescence confirmed that ATOX1/ROCK1 was highly expressed together in Lewis cells. Co-inhibition of ATOX1 and ROCK1 either by siRNA transfection or inhibitory drugs could lead to a significant decrease in tumour proliferation. Interestingly, transcriptional inhibition of ATOX1 can lead to the up-regulation of ROCK1, while inhibition of ROCK1 resulted in the promotion of ATOX1. Moreover, in the analysis of migration ability, a similar synergistic effect from the co-inhibition of ATOX1/ROCK1 was also observed. Finally, the Lewis and Mnu-induced allogeneic transplantation model also demonstrated a greatly improved therapeutic effect by combining targeting ATOX1 and ROCK1. Conclusions: Collectively, our results suggest that a low expression pattern of ATOX1/ROCK1 can predict better clinical outcomes in LUAD. Combining the inhibition of these two targets can reach a significantly better therapeutic effect than targeting either alone. Full article

This study aimed to evaluate how natural dietary supplements, including essential oils (EOs) and probiotics, influence the growth performance, carcass traits, serum components, gut function, gene expression, and jejunal histomorphology of growing quails. A total of 240 unsexed 7-day-old growing Japanese quails were randomly assigned to four experimental groups (n = 60 per group), with each group further divided into six replicates (10 quails per replicate). The control group (S₀) received a basal diet without incorporating any additives, while the experimental groups were supplemented with (i) essential oils (S₁); (ii) probiotics (S₂); or (iii) a mixture of EOs and probiotics (S₃) at a level of 1.5 kg/ton and 0.55 g per kg diet, respectively, and the ratio of the mixture of EOs and probiotics was approximately 2.73:1. The results showed that, from 7 to 35 days of age, S₃ quails showed increased growth performance, carcass weight, and serum total protein with a decreased lipid profile, outperforming the individual supplementation of either additive (p < 0.05). Importantly, EOs or probiotics enhanced immune-antioxidant status in growing quails, particularly those who were fed both EOs and probiotics, showing significantly increased levels of the serum immune parameters IgY and IgM as well as boosting T-AOC, SOD, and GPx levels when MDA content was lowered compared to S₀ quails (p < 0.05). Additionally, in quails fed a mixture of EOs and probiotics, the primary pro-inflammatory factors TNF-α, IL-1β, and IL-6 were downregulated, and the anti-inflammatory factors TGF-β and IL-10 were elevated compared to the S₀ group (p < 0.05). In this context, there was a notable increase in growth (IGF-I, myogenin, and AvUCP), immunological (IL-2, IL-4, IL-6, and AVBD), antioxidant (SOD, CAT, GPx, and ATOX1), and intestinal absorption (VEGF, MUC2, GLUT2, calbindin, and FABP6) markers in quails supplemented with EOs and/or probiotics when compared to the control group (p < 0.05). The combination of EOs and probiotics had the most noticeable impact on the markers’ expression patterns compared to either additive alone (p < 0.05). Consistent with our results, quails given both EOs and probiotics showed significantly greater villi in terms of height and width when compared to the control group in intestinal histomorphology, the primary measure of intestinal wellness. In conclusion, quail diets could benefit from the use of EOs or probiotics as natural growth promoters since they improve growth performance, blood parameters associated with protein and lipid profiles, immune-antioxidant status and inflammation, and marker gene expression profiles of growth, immune, antioxidant, and intestinal absorption. Full article

Background/Objectives: Proper regulation of copper is essential for maintaining neuronal stability and is facilitated by several chaperone proteins, protecting cells from oxidative damage that would otherwise be caused by improperly regulated copper ions. Oxidative stress, resulting from such dysregulation, is hypothesized to play a significant role in the pathogenesis of Alzheimer’s disease (AD). Methods: In this study, we evaluated the concentrations of the copper chaperones CCS, DCTN4, and ATOX1 in control and AD cases via Western blotting and ELISA, and quantified the copper concentrations in fractionated neurons using ICP-MS. Results: Our findings reveal a significant reduction in CCS levels in AD cases (p = 0.0085), with a progressive decline observed with advancing age. This decline was more pronounced in women, although the difference did not reach statistical significance (p = 0.0768). No significant differences were observed in copper concentrations within synaptosomal (p = 0.3869) or cytosolic fractions (p = 0.4461) between the AD and control cases. Additionally, comprehensive analyses of the effects of sex and age showed no significant impact on the levels of copper chaperones or copper distribution across cellular compartments. Conclusions: These results suggest a strong association between reduced CCS levels and AD pathology, highlighting a potential role for CCS in the redistribution of copper ions within neurons. This redistribution may contribute to oxidative stress and neuronal dysfunction, offering new insights into the mechanisms underlying AD pathogenesis. Full article

Research on the effects of organic and inorganic Cu sources on metabolic processes and mechanisms in pigs is lacking. This study investigated the effects of different copper (Cu) sources and levels on hepatic Cu metabolism and transporter factors in growing pigs. Sixty healthy piglets (initial body weight 14.00 ± 0.30 kg) were randomly divided into four groups with five replicates of three pigs each. Four diets (AM, AH, BM, and BH) had different Cu sources [Cu sulphate (CuSO₄): A and Cu amino acids (Cu-AA): B] and levels [supplemented (120 mg/kg DM): M, supplemented (240 mg/kg DM): H]. The pre-feeding period was 7 days, followed by a 45-day feeding period. Slaughter and sample collection were carried out on the 46th day of the formal feeding period. Significant differences were considered at p < 0.05. The final weight and average daily gain (ADG) of growing pigs in the Cu-AA groups were significantly higher than those in the CuSO₄ groups. Serum Cu increased with increasing Cu supplementation on days 20 and 40. Cu concentrations in muscle, liver, and liver subcellular organelles were higher in Cu-AA groups. In the CuSO₄ groups, Cu concentrations were higher in kidneys and faeces. In Cu-AA groups, both the Cu concentrations in lysosomes and cytosol were higher, and the activities of cathepsin D (CTSD), β-glucosidase (BGL), and acid phosphatase (ACP) in lysosomes and cytoplasm were higher. Comparisons between groups showed that liver mRNA of copper transporter protein 1 (CTR1), ATPase copper-transporting beta (ATP7B), ceruloplasmin (CP), antioxidant protein 1 (ATOX1), and metallothionein (MT) was lower in the CuSO₄ group than in the Cu-AA group, with the best performance at 120 mg/kg Cu. mRNAs for ATPase copper-transporting alpha (ATP7A), cytochrome c oxidase copper chaperone 17 (COX17), and copper chaperone for superoxide dismutase (CCS) showed a decreasing trend in the Cu-AA groups. Cu-AA is better for Cu deposition, enhances the utilisation of Cu, reduces Cu excretion, and promotes the expression of relevant enzymes and transporters in the liver. Full article

Background: Multiple myeloma (MM) is a hematologic malignancy characterized by the clonal proliferation of plasma cells, with extramedullary myeloma (EMM) being an aggressive form involving malignant infiltration beyond the bone marrow. Copper metabolism is essential for tumor proliferation and metastasis, with copper metabolism MURR1 domain (COMMD) proteins regulating these processes and maintaining copper homeostasis. Dysregulated copper homeostasis contributes to cancer progression, including MM, with elevated copper levels linked to disease aggressiveness and poor prognosis. This study investigates the role of the COMMD3 in mediating MM cell progression, particularly its influence on copper metabolism. Methods: Comprehensive bioinformatics analyses were conducted on bone marrow and extramedullary samples to determine the expression of COMMD3, which was validated through in vitro and in vivo functional assays. The MM cell lines RPMI8226 and MM1S underwent lentiviral transfection for COMMD3 overexpression and knockdown. RNA sequencing was conducted on COMMD3 knockdown cells to identify differentially expressed genes. Functional assays measured cell proliferation, migration, apoptosis, and copper metabolism, with a non-obese diabetic severe combined immune-deficiency gamma (NSG) mouse xenograft model providing in vivo validation. Results: Elevated COMMD3 expression was correlated with extramedullary myeloma and poor prognosis in MM patients. COMMD3 promoted MM cell proliferation and migration, modulating intracellular copper levels, likely through the ATOX1-ATP7A-LOX copper-metabolism-related pathway. High ATOX1 expression was correlated with worse outcomes, and ATOX1 inhibition abolished COMMD3’s effects. Conclusions: This study highlights the pivotal role of COMMD3 in MM progression, particularly via the ATOX1-ATP7A-LOX axis. These findings provide insights into EMM mechanisms and position COMMD3 as a potential therapeutic target. Future research is needed to validate these findings in larger clinical cohorts and to unravel the precise molecular interactions between COMMD3 and copper metabolism proteins. Full article

Arthritis is a leading cause of economic loss in livestock farming including sheep. This study examined the changes in gene expression, antioxidants, pro-inflammatory cytokines, acute-phase proteins (APPs), hormonal assays and iron profiles linked to sheep arthritis, as well as the diagnostic utility of these markers. Blood samples were obtained from 30 apparently healthy rams and 30 rams with arthritis for gene expression and biochemical analyses. Gene expression intensities were much higher in the arthritis-affected rams than in the healthy ones for the genes IL-1α, IL-1β, IL-6, IL-10, TNFα, NCF4, NFKB, TMED, FCAMR, iNOS and COX18. The SOD3, CAT, GPX and ATOX1 genes were expressed at substantially lower levels in arthritis-affected rams. Disparities in the nucleotide sequence variants for the amplified DNA bases linked to arthritis for the studied genes were found in the PCR-DNA sequence verdicts of the affected and healthy rams. Immunological, acute-phase protein (APP), antioxidant, hormonal and iron profiles were estimated in both groups and statistically analyzed. The arthritic group in relation to the healthy one showed a significant (p < 0.05) increase in pro-inflammatory cytokines, APPs, free radicals, immunoglobulins, cortisol, GH, TSH, ferritin, TIBC and UIBC and a significant (p ˂ 0.05) decrease in anti-inflammatory cytokines, antioxidants, complements, insulin, T3, T4, SI, and Tf and Tf sat.% serum levels. The estimated pro-inflammatory cytokines and APPs achieved high values of sensitivity and specificity, positive predictive values (PPVs), negative predictive values (NPVs), a high accuracy rate and a moderate likelihood ratio (LR). The study concluded that ovine arthritis stimulates innate and humeral immunity, resulting in prominent alterations in gene expression, pro-inflammatory cytokines, APP assays and antioxidant profiles, which could be valuable indicators of sheep arthritis. Full article

Neurodegenerative diseases have been increasingly plaguing the global population, with attempts to understand their etiopathogenesis and pursue therapeutics being at the forefront of multidisciplinary efforts. To that end, research was launched in our lab, based on natural products and bioessential metal ion complex forms to peruse their antioxidant and neuroprotective potential at the cellular level. To that end, the bioactivity profile of optimized Cornus mas L. extracts and supplemented mixtures thereof with soluble-bioavailable well-characterized hybrid materials, Zn(II)-Cit and V(IV)-Cit, was investigated. In vitro experiments on sensitive brain tissue cell lines (N2a58, SH-SY5Y) showed that the extracts and the metal complexes were atoxic (morphology, proliferation, chemotacticity) in a concentration-dependent manner. Subsequently, the antioxidant potential of all materials was examined, with H₂O₂ as the oxidizing agent, thereby revealing through viability and reactive oxygen species (ROS) visualization significant antioxidant activity, while specific genes (NFE2L2, Hmox1, GCLM) were crucial in divulging mechanistic aspects of the antioxidation. Concurrently, the anti-inflammatory activity was evaluated through gene expression (TNF-a, IL-6), with Zn(II) bioavailability projecting intracellular levels linked to the observed sustainable activity. The collective bioactivity profile of the extracts and Zn(II)-Cit reveals significant neuroprotective properties, thereby meriting development of new naturally-based neutraceuticals that proactively avert neuropathological aberrations. Full article

Tetanus neurotoxins (TeNT) and botulinum neurotoxins (BoNTs) are closely related ~150 kDa protein toxins that together comprise the group of clostridial neurotoxins (CNTs) expressed by various species of Clostridia. While TeNT is expressed as a single polypeptide, BoNTs are always produced alongside multiple non-toxic proteins that form a stabilizing complex with BoNT and are encoded in a conserved toxin gene cluster. It is unknown how tent evolved without a similar gene cluster and why complex-free TeNT is secreted as a stable and soluble protein by C. tetani, whereas complexing proteins appear to be essential for BoNT stability in culture supernatants of C. botulinum. To assess whether the stability of TeNT is due to an innate property of the toxin or is a result of C. tetani’s intra- and extra-cellular environment, both TeNT and complex-free BoNT/A1^ERY were expressed recombinantly in atoxic C. tetani and analyzed for expression and stability. The strong clostridial ferredoxin (fdx) promotor resulted in the expression of recombinant TeNT at greater levels and earlier time points than endogenously produced TeNT. Recombinant BoNT/A1^ERY was similarly expressed by atoxic C. tetani, although partial degradation was observed. The rBoNT/A1^ERY produced in C. tetani was also partially proteolytically processed to the dichain form. Investigations of bacterial growth media and pH conditions found that the stability of rTeNT and rBoNT/A1^ERY in spent media of C. tetani or C. botulinum was affected by growth media but not by pH. These data indicate that the distinct metabolism of C. tetani or C. botulinum under various growth conditions is a primary factor in creating a more or less favorable environment for complex-free CNT stability. Full article

Recombinant mutant holotoxin BoNTs (rBoNTs) are being evaluated as possible vaccines against botulism. Previously, several rBoNTs containing 2–3 amino acid mutations in the light chain (LC) showed significant decreases in toxicity (2.5-million-fold–12.5-million-fold) versus wild-type BoNT/A1, leading to their current exclusion from the Federal Select Agent list. In this study, we added four additional mutations in the receptor-binding domain, translocation domain, and enzymatic cleft to further decrease toxicity, creating 7M rBoNT/A1. Due to poor expression in E. coli, 7M rBoNT/A1 was produced in an endogenous C. botulinum expression system. This protein had higher residual toxicity (LD50: 280 ng/mouse) than previously reported for the catalytically inactive rBoNT/A1 containing only three of the mutations (>10 µg/mouse). To investigate this discrepancy, several additional rBoNT/A1 constructs containing individual sets of amino acid substitutions from 7M rBoNT/A1 and related mutations were also endogenously produced. Similarly to endogenously produced 7M rBoNT/A1, all of the endogenously produced mutants had ~100–1000-fold greater toxicity than what was reported for their original heterologous host counterparts. A combination of mutations in multiple functional domains resulted in a greater but not multiplicative reduction in toxicity. This report demonstrates the impact of production systems on residual toxicity of genetically inactivated rBoNTs. Full article

Atomic oxygen (AtOx) is a major component of the space environment between 200 and 800 km (LEO—low Earth orbit region) and is the principal source of erosion for exposed aerospace structures. The damage to surface materials is proportional to the AtOx fluence, which depends on altitude, exposure time, orbital inclination, and solar activity, and it is caused by the formation of volatile oxides which do not adhere to the surface; furthermore, the mass loss may also be worsened by UV radiation, which increases the chemical degradation of the exposed material. Carbon/carbon (C/C) is an advanced ceramic composite that is frequently found as a base component of thermal protection systems (TPS), rocket nozzles, or other spacecraft subsystems. In this work, a simulation of the AtOx/UV synergistic effects on C/C plates exposed at different attitude positions were carried out by experimental tests performed at the Aerospace Systems Laboratory (LSA—Sapienza University of Rome) by means of an Atomic Oxygen OS-Prey RF plasma source, which also included a high-power UV-ray generator. The present experimental plan was built on the activity developed during recent years at LSA concerning the study of C/C materials for protecting aerospace structures from thermal shock in re-entry missions. The tests were conceived by considering a fixed time of exposure with a base fluence of 7.6 × 10¹⁹ n.s./cm², as evaluated from the erosion of the reference samples exposed to AtOx flux at a normal incidence; the simulation of the different attitude positions was then analyzed, also considering the simultaneous effect of UV radiation. The results of the aging ground test suggest the following: (i) C/C oxidation in LEO must be taken into full consideration in the TPS design with reference to protective coating solutions, (ii) the LEO environment simulation is closely related to AtOx/UV combined irradiation, as well as to the spacecraft’s in-orbit attitude. Full article

In the field of human health research, the homeostasis of copper (Cu) is receiving increased attention due to its connection to pathological conditions, including diabetes mellitus (DM). Recent studies have demonstrated that proteins associated with Cu homeostasis, such as ATOX1, FDX1, ATP7A, ATPB, SLC31A1, p53, and UPS, also contribute to DM. Cuproptosis, characterized by Cu homeostasis dysregulation and Cu overload, has been found to cause the oligomerization of lipoylated proteins in mitochondria, loss of iron–sulfur protein, depletion of glutathione, production of reactive oxygen species, and cell death. Further research into how cuproptosis affects DM is essential to uncover its mechanism of action and identify effective interventions. In this article, we review the molecular mechanism of Cu homeostasis and the role of cuproptosis in the pathogenesis of DM. The study of small-molecule drugs that affect these proteins offers the possibility of moving from symptomatic treatment to treating the underlying causes of DM. Full article

Background: Cuproptosis induction is seen as a promising alternative for immunotherapies and targeted therapies in breast cancer. The objective of this research was to examine the prognostic and biological importance of cuproptosis-related genes (CRGs) in lung adenocarcinoma (LUAD). Methods: The following methods were used: GSE10072 dataset and TCGA database analysis, differential expression analysis of CRGs, and biological function (BP) and signaling pathway enrichment analysis, prognostic analysis and clinical analysis of CRGs, construction of the prognostic signature and RNA modified genes and miRNA analysis of CRGs in LUAD, immunoinfiltration analysis and immunohistochemical staining of DβH, UBE2D3, SOD1, UBE2D1 and LOXL2. Results: AOC1, ATOX1, CCL8, CCS, COX11, CP, LOXL2, MAP2K2, PDK1, SCO2, SOD1, UBE2D1, UBE2D3 and VEGFA showed significantly higher expression, while ATP7B, DβH, PDE3B, SLC31A2, UBE2D2, UBE2D4 and ULK2 showed lower expression in LUAD tissues than normal tissues. We also found that ATP7B (4%), AOC1 (3%) PDE3B (2%), DβH (2%), CP (1%), ULK2 (1%), PDK1 (1%), LOXL2 (1%) and UBE2D3 (1%) showed higher mutation frequencies. The univariate Cox analysis was used to identify CRGs that have prognostic value. It identified 21 genes that showed significant prognostic value, containing DβH, UBE2D3, SOD1, UBE2D1 and LOXL2. Patients with DβH up–expression have a longer survival time and patients with UBE2D3, SOD1, UBE2D1 and LOXL2 down–expression also have a longer survival time. hsa–miR–29c–3p, hsa–miR–29a–3p, hsa–miR–181c–5p, hsa–miR–1245a, etc., play an important role in the miRNA regulatory network, and in LUAD, miR–29a, miR–29c and miR–181c high expression survival was longer, and miR–1245a low expression survival was longer. We also performed an analysis to examine the relationships between DβH, LOXL2, SOD1, UBE2D1 and UBE2D3 and immune infiltration in LUAD, including B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils and DCs. Conclusion: DβH, UBE2D3, SOD1, UBE2D1, and LOXL2 are potential candidates implicated in LUAD and can be further explored for their application as diagnostic, prognostic, and therapeutic biomarkers for LUAD. Full article

Vaccines are one of the most effective strategies to prevent pathogen-induced illness in humans. The earliest vaccines were based on live inoculations with low doses of live or related pathogens, which carried a relatively high risk of developing the disease they were meant to prevent. The introduction of attenuated and killed pathogens as vaccines dramatically reduced these risks; however, attenuated live vaccines still carry a risk of reversion to a pathogenic strain capable of causing disease. This risk is completely eliminated with recombinant protein or subunit vaccines, which are atoxic and non-infectious. However, these vaccines require adjuvants and often significant optimization to induce robust T-cell responses and long-lasting immune memory. Some pathogens produce protein toxins that cause or contribute to disease. To protect against the effects of such toxins, chemically inactivated toxoid vaccines have been found to be effective. Toxoid vaccines are successfully used today at a global scale to protect against tetanus and diphtheria. Recent developments for toxoid vaccines are investigating the possibilities of utilizing recombinant protein toxins mutated to eliminate biologic activity instead of chemically inactivated toxins. Finally, one of the most contemporary approaches toward vaccine design utilizes messenger RNA (mRNA) as a vaccine candidate. This approach was used globally to protect against coronavirus disease during the COVID-19 pandemic that began in 2019, due to its advantages of quick production and scale-up, and effectiveness in eliciting a neutralizing antibody response. Nonetheless, mRNA vaccines require specialized storage and transport conditions, posing challenges for low- and middle-income countries. Among multiple available technologies for vaccine design and formulation, which technology is most appropriate? This review focuses on the considerable developments that have been made in utilizing diverse vaccine technologies with a focus on vaccines targeting bacterial toxins. We describe how advancements in vaccine technology, combined with a deeper understanding of pathogen–host interactions, offer exciting and promising avenues for the development of new and improved vaccines. Full article