Biology subjects

Qin, J.

Publications and source records attributed to Qin, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Mechanistic modeling of bacterial translation initiation across growth conditions

Translation frequency in bacteria depends on how ribosomes, mRNAs, and initiation factors are allocated across growth conditions. Here, we developed a mechanistic ODE-based model of Escherichia coli translation that represents initiation, elongation, termination, and coupled auxiliary processes. Growth-dependent abundances were derived from physiological relationships and reprocessed omics data, and simulated outputs were compared with translation-frequency and active-ribosome references. The model predicts a continuous shift from complex-formation-limited toward ribosome-limited behavior as growth increases. This shift is characterized by a decline in free-ribosome abundance, whereas initiation-factor pools remain largely unbound and do not become depleted in parallel. Together with the implemented IF-dependent kinetic term, this preserved availability provides a model-internal route through which productive initiation can be maintained despite increasing ribosome utilization. Consistently, transcript-wide ribosome loading remains below its theoretical maximum, while COG-level simulations reveal distinct sector-specific translation-frequency trajectories. The study therefore provides a resource-allocation framework for interpreting how mRNA--ribosome interactions shape bacterial translation across growth conditions.

systems biology↗

bioRxiv2026

Genome-wide screen for genes required for smooth lipopolysaccharide production in Escherichia coli K-12

Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria, contributing to membrane integrity and environmental interactions. Genome-wide studies defining bacterial gene functions have been extensively performed in the model strain Escherichia coli K-12 which lacks O-antigen (OAg), and therefore does not produce smooth LPS (S-LPS). Consequently, the genetic requirements for S-LPS production in this model system remain incompletely defined. Here, a functional wbbL gene was introduced into the E. coli K-12 KEIO single-gene deletion mutant library to restore OAg synthesis, enabling genome-wide analysis of S-LPS production by screening with colicin E2 (ColE2) and validated with LPS silver staining. This identified 319 mutants with increased sensitivity to ColE2 in the presence of OAg, suggesting broader envelope-associated effects during screening. In addition, 27 mutants showed defects in S-LPS production, corresponding to genes involved in OAg biosynthesis, LPS core and sugar precursor synthesis, OAg ligation and regulation, and enterobacterial common antigen biosynthesis. A further 18 mutants initially appeared defective in S-LPS production but could not be validated upon reconstruction, and whole-genome sequencing revealed secondary mutations responsible for the observed phenotypes. This study provides a validated genetic framework for S-LPS production in E. coli K-12 and highlights the importance of rigorous validation in genome-wide screening approaches.

microbiology↗

bioRxiv2026

An engineered IdeS variant with enhanced activity and performance for IgG degradation

IgG-degrading enzymes have emerged as innovative therapeutic agents for treating conditions driven by pathogenic antibodies. Here, we used structure-guided rational design to engineer IdeSM33, a double mutant (K167R/D226E) of the IgG-specific bacterial protease IdeS from Streptococcus pyogenes, with improved catalytic efficiency. Biolayer interferometry revealed a fourfold increase in binding affinity relative to wild-type IdeS (IdeSWT). This enhancement is likely attributable to mutations that strengthen hydrogen bonding at the enzyme-IgG Fc interface. In vitro, IdeSM33 has higher performance than IdeSWT in cleaving serum IgG. In vivo studies in rabbits demonstrated that IdeSM33 effectively depleted circulating IgG and showed better performance at a dose of 0.005 mg/kg than the IdeSWT. Although doses greater than 0.2 mg/kg demonstrated higher plasma concentrations of IdeS and a larger AUClast, they did not show a significant enhancement in the pharmacodynamics of IgG degradation. Importantly, a single dose of IdeSM33 (0.2 mg/kg) potently degraded binding and neutralizing antibodies against AAV9 within 1-2 days and restored hepatic AAV9 transduction in pre-immunized animals. Together, these findings highlight IdeSM33 as a potent and safe engineered enzyme with therapeutic potential for autoimmune disorders, transplant rejection, and overcoming pre-existing humoral immunity in gene therapy.

bioengineering↗

bioRxiv2026

Chronic Stress Alters Dorsal Bed Nucleus of Stria Terminalis Synaptic Neurotransmission in a Dravet Syndrome Mouse Model

Stress is a commonly reported seizure precipitant and may contribute to the development of psychiatric comorbidities in epilepsy, yet how chronic stress interacts with epileptic circuits remains poorly understood. We investigated the impact of chronic restraint stress on physiological, behavioral, and synaptic outcomes in a mouse model of Dravet syndrome, specifically corticotropin-releasing factor (CRF) neurons in the bed nucleus of the stria terminalis (BNST), a stress-responsive region implicated in epilepsy patients. Chronic restraint stress produced divergent hypothalamic-pituitary-adrenal axis responses, with stressed Dravet syndrome mice exhibiting elevated corticosterone, increased mortality in females, and increased locomotion and anxiety-like behavior. Ex vivo electrophysiological recordings revealed that chronic stress increased spontaneous excitatory event frequency onto BNST CRF neurons in both genotypes and selectively increased sEPSC and sIPSC amplitude in Dravet syndrome mice. Evoked recordings demonstrated genotype-specific effects of stress on glutamatergic transmission in CRF neurons of the DS group. This suggests greater stress-dependent remodeling of spontaneous and evoked synaptic activity in DS. These findings suggest chronic stress may worsen physiological and behavioral outcomes in Dravet syndrome and promote specific maladaptive alterations in BNST CRF circuitry. More broadly, these results suggest that stress interacts with seizure vulnerability and potentially contributes to neuropsychiatric comorbidities and epilepsy.

neuroscience↗

bioRxiv2026

Bridging genomes and peptidomes: hybrid sequencing reveals conserved bioactive peptides in crustaceans

Endogenous peptides are critical regulators of signaling and immunity but remain difficult to characterize in organisms with incomplete genomic annotation. We developed a hybrid discovery platform that integrates transformer-based de novo sequencing (Casanovo), neuropeptide-focused database searching (EndoGenius), and empirical false discovery rate estimation via NovoBoard. This pipeline enables confident identification of endogenous peptides while expanding coverage beyond conventional database-only or de novo-only approaches. Applied to neuroendocrine tissues from Callinectes sapidus and Cancer borealis, the workflow revealed numerous high-abundance novel peptides and provided structural and genomic support for their biological relevance. Notably, we report the first histone-2A-derived antimicrobial peptide in the C. sapidus and characterize naturally occurring sequence variants. We also identified unexpected peptide homologies between crustaceans and Rattus norvegicus, enabling annotation of conserved housekeeping proteins in sparsely annotated genomes. This hybrid platform establishes a scalable, open-source strategy for advancing neuropeptidomics and endogenous peptide discovery in emerging model organisms.

bioinformatics↗

bioRxiv2026

Clarified an rDNA Gene Unit Pattern with (CTTT)n and (CT)n Microsatellites Aggregation Ahead of and Behind the Gene in Human Genome

BackgroundConventional models of human ribosomal DNA (rDNA) array organization have historically depended on transcription-centric boundaries, partitioning the unit into a [~]13 kb rDNA transcription region and a monolithic [~]31 kb intergenic spacer (IGS). While our previous identification of Duplication Segment Units (DSUs) mapped these arrays based on an intuitive analysis of the microsatellite density landscape of the complete reference human genome, our present deep mining of this landscape has revealed a more accurate rDNA Gene Unit Pattern. Methods & ResultsIn this study, we conducted a deep mining analysis of our previously established microsatellite density landscape of the T2T-CHM13 assembly, focusing specifically on nucleolar organizing regions (NORs). We suggest a more accurate rDNA Gene Unit Pattern containing a (CTTT)n microsatellite aggregation ahead of the rDNA gene and a (CT)n microsatellite aggregation behind the gene, rather than a pattern featuring an IGS region inserted between two rDNA genes. ConclusionsA correct rDNA gene pattern of the human genome probably includes a (CTTT)n microsatellite aggregation ahead of the gene and a (CT)n microsatellite aggregation behind it, which possibly constitute cis- and trans-regulating regions; the (CTTT)n and (CT)n microsatellite aggregations may provide two different local stable DNA structures for regulatory protein binding.

genetics↗

bioRxiv2026

Subchondral H-type Blood Vessel Formation Aggravates Articular Cartilage Degeneration through LEP-LEPR Axis

BackgroundOsteoarthritis (OA) is a chronic degenerative joint disease characterized by cartilage deterioration that has resulted in severe physical and economic costs for society. Risk factors for osteoarthritis encompass genetic predisposition, mechanical influences, obesity, inflammation, and metabolic problems. Recent reports indicate that the morphological alterations of subchondral bone evolve dynamically with the advancement of osteoarthritis. A significant alteration in subchondral bone is the development of aberrant blood vessels, particularly H-type vessels. H-type arteries are essential for preserving subchondral bone homeostasis. They support nutrients and oxygen to bone tissue and eliminate waste, thereby supporting the metabolic activities of osteoblasts and osteoclasts. However, the upstream factors governing H-type arteries are yet unknown. MethodsThe whole transcriptome analysis was performed by informatics which is further verified in clinical samples. Using LEP and LEPR knockout mice, the correlation between abnormal subchondral bone metabolism and LEP, LEPR, and H-type vessel formation were uncovered. ConclusionsThis study reveals that the lipid metabolism pathway was highly enriched on the OA lesion, and LEP and LEPR were markedly underexpressed in the subchondral bone on the side with severe OA lesions. The causal relationship of the LEP-LEPR-CD31 signalling pathway in aging and osteoarthritis models is revealed at the overall and molecular levels. This project will provide a new theoretical basis for the pathogenesis of early OA and new ideas and targets for the clinical treatment of OA by exploring the LEP-LEPR-CD31 signalling in osteoarthritis.

pathology↗

bioRxiv2025

Transcriptional profiling of Pseudomonas aeruginosa biofilm life cycle stages reveals dispersal-specific biomarkers

Bacteria exhibit two lifestyles: planktonic free-floating individual cells or sessile multicellular aggregates known as biofilms. The biofilm lifecycle is characterised by three distinct stages: attachment, maturation and dispersal. Distinct adaptations occur in each stage, determining cellular behaviours such as surface attachment or synthesis and degradation of extracellular matrix components. Characterising stage-specific bacterial profiles therefore represents a valuable strategy for the development of novel antibiofilm therapies. Here, we used the model biofilm-forming bacterium Pseudomonas aeruginosa PAO1 to characterise the transcriptional profiles of each stage of the biofilm life cycle: attachment, biofilm maturation and spontaneous dispersal in closed cultures. We report that surface attachment was accompanied by the upregulation of genes comprising the Pil-Chp mechanosensory system, whereas biofilm maturation was characterised by the upregulation of genes involved in Pel polysaccharide synthesis, siaD and PA4396 diguanylate cyclases as well as pipA, fimX and PA5442. In contrast, dispersing cells upregulated genes responsible for the biosynthesis of alginate, rhamnolipid, and extracellular nucleases (eddA, eddB), as well as the transcriptional regulator of dispersal amrZ. Additionally, genes encoding the spontaneous dispersal molecule cis-2-decenoic acid (dspS and dspI), canonical phosphodiesterases (nbdA and rbdA), four non-canonical HD-GYP phosphodiesterases and seven other c-di-GMP-related enzymes were also upregulated during dispersal. Our comprehensive analysis of transcriptional changes across biofilm stages therefore provides benchmarking stage-specific transcriptional profiles for P. aeruginosa biofilms in closed culture systems. Furthermore, it allowed the identification of a subset of fourteen genes as transcriptional biomarkers of dispersal, which were used to build reporter plasmids as tools to determine the onset of dispersal. ImportanceBiofilm infections by P. aeruginosa are a major medical challenge due to the increased tolerance to antimicrobials displayed by bacteria living in sessile communities, which is reduced during spontaneous biofilm dispersal. Attachment, biofilm maturation and dispersal represent the main stages of a dynamic process known as the biofilm lifecycle. However, the global regulatory responses governing transitions between these stages remain understudied. Here, we combine live microscopy and biomass quantification to track the progression of P. aeruginosa cultures through the three main stages of the biofilm lifecycle. We show that cells from each stage recapitulate canonical, stage-specific transcriptional responses and identify a set of biomarkers associated with the onset of dispersal. These biomarkers may offer a practical tool for rapidly screening dispersal-inducing compounds, aiding in the discovery of the next generation of antibiofilm therapeutics.

genomics↗

bioRxiv2025

AAV vectors with enhancer-controlled ITR promoters

Adeno-associated virus (AAV) is limited by its packaging capacity and the unwanted promoter activity of inverted terminal repeats (ITRs). Here we utilized mini-enhancers to regulate ITR promoters and drive AAV cargo expression in the absence of the canonical promoters, which not only solved the ITR issue but also released more payload for the cargo. As an example, this new design successfully enabled robust, tissue-specific SpCas9 gene editing via a single AAV, which otherwise requires a dual-AAV system due to the large size of SpCas9.

bioengineering↗

bioRxiv2025

PPARgene 2.0: leveraging large language models and multi-omics data for enhanced identification and prediction of PPAR target genes

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors of the nuclear receptor superfamily. Upon ligand binding, PPARs activate target gene transcription and regulate a variety of important physiological processes such as lipid metabolism, inflammation, wound healing and immune responses. PPARgene is a database that integrates literature-curated and computationally predicted PPAR target genes. It provides gene-level annotations including tissue specificity, species, and supporting PubMed IDs. Computational predictions are generated using a machine learning method that combines PPRE motif analysis with microarray expression data. Here, we introduce PPARgene 2.0, a 10-year update to the original PPARgene database. This update adds 35 newly reported PPAR target genes, 20 PPAR{beta}/{delta} target genes, and 72 PPAR{gamma} target genes, bringing the total number of curated target genes in the database to 337. To retrieve newly reported PPAR target genes from the literature, we used two language models to screen publications after 2016. Candidate papers were then manually reviewed, and verified target genes were added to the updated database. This update also improved the predictive method by expanding the volume of high-throughput gene expression data and incorporating PPAR-related ChIP-seq datasets alongside in silico PPRE analysis. Fivefold cross-validation demonstrated that the new predictive method outperforms the original one. The updated prediction tool is available as part of the PPARgene 2.0 platform. The database is openly accessible at https://www.ppargene.org.

bioinformatics↗

bioRxiv2025

Disease mutations in the N-terminal extension and α 2'-helix of dyskerin do not abolish dimerization but disrupt binding to the telomerase RNA

The H/ACA ribonucleoprotein complex component dyskerin is essential for the biogenesis of H/ACA RNAs, including the human telomerase RNA (hTR). The N-terminal extension and 2 helix of dyskerin are hotspots for disease-associated mutations linked to X-linked dyskeratosis congenita (X-DC), a premature aging disorder. Some of these mutations disrupt dyskerin-hTR interactions, leading to hTR destabilization and reduced telomerase activity. Cryo-EM structures of human telomerase have shown that the N-terminal extension and 2 helix participate in dyskerin dimerization. However, biochemical evidence for dyskerin dimerization is still lacking, and it remains unclear whether mutations in these regions impair hTR binding by disrupting dimerization. Here, we provide the first biochemical evidence that dyskerin undergoes dimerization. We further demonstrate that dimerization is RNA independent and not abolished by disease mutations in the N-terminal extension or 2 helix. Instead, these mutations impair hTR binding. Our findings offer new mechanistic insight into how mutations in the dyskerin N-terminal extension and 2 helix contribute to the pathogenesis of X-DC.

cell biology↗

bioRxiv2025

MSF-CPMP: A Novel Multi-Source Feature Fusion Model for Prediction of Cyclic Peptide Membrane Permeability

Cyclic peptides are becoming attractive molecules for drug discovery because of their properties with inherent stability and structural diversity. However, the high potential of cyclic peptide drugs is challenged by the limited membrane permeability cross cell membrane. To predict cyclic peptide membrane permeability (CPMP), an increased number of computational models or tools are designed and used. But these existing algorithms or models do not appropriately capture feature diversity of cyclic peptides. In this study, we introduce a novel multi-source feature fusion model called MSF-CPMP, which aims to increase the accuracy of predicted CPMP. The MSF-CPMP model incorporates three features extracted from SMILES sequences, graph-based molecular structures, and physicochemical properties of cyclic peptides. By benchmarking with other machine learning and deep learning-based methods, MSF-CPMP achieved the highest levels of the evaluation metrics such as accuracy of 0.9062 and AUROC of 0.9546, and further validated MSF-CPMP robustness in learning capabilities and efficacy of its multi-source fusion. Our result demonstrates that MSF-CPMP outperforms other methods in predicting CPMP, that provides also exemplifies the power of advanced deep learning methods in tackling complex biological challenges, offering contributions to computational biology and clinical treatment. Code is available at https://github.com/wanglabhku/MSF-CPMP Author summaryWe have recognized that cyclic peptides are one type of main macro-molecular drugs useful for treatment of human diseases. However, the structural diversity of cyclic peptides results in limited permeability across cell membranes which is challenging drug research or industrial development. Previously some computational models or tools including machine learning or deep learning ones tried to solve this problem, but they still are not efficient. There are urgent requirement for more effective methodologies. Thus, we introduce a novel multi-source feature fusion model called MSF-CPMP. Our aim is to enhance accuracy and efficiency of predicted CPMP. We carried out evaluation of performance metrics by comparative analysis between MSF-CPMP and other machine learning or deep learning methods. Our result demonstrates that MSF-CPMP outperforms other models in predicting CPMP. We further validated MSF-CPMP robustness in learning capabilities and efficacy of its multi-source feature fusion. In the future we would further improve our methods that can integrate broader biomedicine and biomedical information and provide guide for drug diversity in clinically treating complex human diseases.

bioinformatics↗

bioRxiv2025

Nitric oxide donor sodium nitroprusside serves as a source of iron supporting Pseudomonas aeruginosa growth and biofilm formation

Biofilm dispersal agents, like nitric oxide (NO), restore antimicrobial effectiveness against biofilm infections by inducing bacteria to shift from a biofilm to a planktonic state, thereby overcoming the antimicrobial tolerance typically associated with biofilms. Sodium nitroprusside (SNP) is a widely used NO donor for investigating the molecular mechanisms underlying NO-mediated biofilm dispersal in the nosocomial pathogen Pseudomonas aeruginosa. However, the biofilm effects of SNP are variable depending on the in vitro experimental conditions, with some studies reporting enhanced growth in both planktonic and biofilm forms instead of dispersal. These discrepancies suggest that SNP affects P. aeruginosa biofilm-residing cells beyond the release of NO. In this study, we compared SNP with another NO donor, Spermine NONOate, to systematically contrast their effects on biofilm and planktonic cultures of P. aeruginosa. We found that SNP, but not Spermine NONOate, increased the biomass of P. aeruginosa biofilms in microplate cultures. This effect was also observed when biofilms were supplemented with iron. Additionally, supplementation with SNP rescued the planktonic growth of P. aeruginosa in iron-depleted media, similar to FeSO4 supplementation, suggesting that SNP may serve as an iron source. Our findings suggest that SNPs potential as an NO agent used for biofilm dispersal may be confounded by its role in promoting both biofilm and planktonic growth through its iron centre. Our study cautions investigators using SNP for studying NO-mediated biofilm dispersal.

microbiology↗

bioRxiv2025

Disruptions in Outer Membrane-Peptidoglycan Interactions Enhance Bile Salt Resistance in O-antigen-Producing E. coli

Bile salts (BS) are derived from cholesterol in the liver and act as antimicrobial agents in the intestines by disrupting bacterial cell membranes and inducing oxidative stress. The gut bacterium E. coli is naturally resistant to BS, including the model strain K12 that produces a truncated LPS without O-antigen (OAg). Paradoxically, restoring a wild-type like LPS with OAg sensitises E. coli K12 to exogenous BS. In this study, we investigate this phenomenon. We show that mutations causing truncation of the LPS core oligosaccharide render these strains even more susceptible to BS, similar to the mutant strain MG1655-S{Delta}waaL defective in OAg ligase, primarily due to the accumulation of the lipid-linked intermediate UndPP-OAg. Through the characterisation of BS-resistant suppressor mutants of MG1655-S{Delta}waaL, we identify key genetic disruptions involved in resistance. Notably, we observed the highest BS resistance in strains with a weaker connection between the outer membrane (OM) and peptidoglycan (PG), including strains lacking the major OM-anchored, PG-binding proteins OmpA or Lpp, or expressing versions of these that lack PG-binding. Our data suggest that BS-induced stress in OAg-producing E. coli is due to the spatial constraints between OM and PG, and that mutations disrupting OM-PG interactions alleviate this stress, enhancing BS resistance. These findings provide new insights into a major challenge E. coli faces in the gut environment where it needs to produce OAg for stable colonisation and resists BS. E. coli can only survive BS exposure by fine-tuning the connectivity between its cell envelope layers, which highlights a potential target for modulating bacterial responses to BS in the gut. Author summaryEnteric bacteria residing in the human gut must withstand the host-derived antimicrobial agents, bile salts (BS), but the underlying resistance mechanisms are not fully elucidated. This study investigates the BS resistance mechanisms in O-antigen (OAg)-producing Escherichia coli K-12. We show that truncation of lipopolysaccharide (LPS) core oligosaccharides or restoration of OAg production increases BS sensitivity due to the accumulation of UndPP-OAg intermediates. By analysing suppressor mutants, we identify key genetic disruptions, particularly affecting the level of outer membrane-peptidoglycan (OM-PG) interactions involving OmpA and Lpp, which confer heightened BS resistance. Our findings highlight how BS-induced stress is linked to spatial constraints between the OM and PG layer, offering new insights into bacterial adaptation to BS stress. This research may provide new targets for therapeutic interventions to modulate gut microbial responses to BS.

microbiology↗

bioRxiv2025

Radiation exposure induced blood-brain barrier injury via mitochondria-mediated sterile inflammation

Radiation-induced brain injury (RIBI) is caused by exposure to high doses of ionizing radiation and characterized by severe cognitive dysfunction and brain necrosis. The widespread application of radiotherapy and rapid development of deep space exploration have substantially increased the risk of RIBI. However, the pathogenesis of RIBI is not fully understood, and no effective intervention is available. This work described a blood-brain barrier (BBB) microphysiological system (MPS), that allowed to explore the responses of BBB and distinct brain cells to radiation exposure. This BBB MPS can recapitulate the interface structure and function of BBB in brain microenvironment, including brain endothelial cells, astrocytes, pericytes and microglia co-cultured under flow condition. Following acute exposure to radiation of X-ray or {gamma}-ray, characteristic RIBI-associated pathological responses were observed, including BBB compromise, DNA breaks, inhibited cell proliferation, cell hypertrophy and pro-inflammatory cytokine release. Among the distinctive types of cells, brain endothelial cells showed the highest radiosensitivity as compared to other cells in the MPS. Intriguingly, X-ray and {gamma}-ray radiation consistently induced prominent sterile inflammation responses, especially type I interferon response, in the BBB MPS. These responses were mediated by radiation-induced mitochondrial DNA release and subsequent activation of cGAS-STING signaling pathway. Furthermore, we found abrocitinib (JAK1 inhibitor) and idebenone (mitochondrial protectant) can attenuate radiation-induced inflammation and ameliorate BBB injury. These findings revealed the involvement of mitochondria-mediated inflammation in RIBI pathogenesis, identifying mitochondria as a potential target for new radioprotective measures.

neuroscience↗

bioRxiv2025

Optimal membrane fluidity modulates biofilm formation in Escherichia coli

IMPACT STATEMENTMembrane fluidity plays a crucial role in bacterial fitness and adaptation to cope rapid environmental changes. While high membrane fluidity promotes robust biofilm formation in Klebsiella pneumoniae, studies in several other species, including Salmonella enterica, suggest that biofilm formation is associated with reduced fluidity. This paradox may reflect the complex relationship between lipid composition and biofilm formation. Our findings demonstrated that both low and high extremes of lipid fluidity restrict biofilm formation. We propose that the required fluidity for biofilm growth, relative to that required for planktonic growth, may differ between species and are readily adjusted during lifestyle transitions.

microbiology↗

bioRxiv2024

Selection, Design and Immunogenicity Studies of ASFV Antigens for Subunit mRNA Cocktail Vaccines with Specific Immune Response Profiles

Development of safe and effective subunit vaccines for controlling African Swine Fever Virus (ASFV) infection has been hampered by a lack of protective viral antigens, complex virion structures, and multiple mechanisms of infection. Here, we selected ASFV antigens based on their localization on the virion, known functions, and homologies to the subunits of the protective vaccinia virus vaccine. We also engineered viral capsid proteins for inducing optimal antibody responses and designed T cell-directed antigen for inducing broad and robust cellular immunity. The selected antigens in lipid nanoparticle-mRNA formulations were evaluated for immunogenicity in both mice and pigs with concordant results. Different antigens induced divergent immune response profiles, including the levels of IgG and T cell responses and effector functions of anti-sera. We further developed a computational approach to combine antigens into cocktails for inducing specific immune response profiles and validated candidate cocktail vaccines in mice. Our results provide a basis for further evaluating candidate subunit mRNA vaccines in challenge studies. TeaserNovel strategies to develop subunit vaccines for ASFV and other complex large DNA viruses.

immunology↗

bioRxiv2024

Membrane Expression Enhances Folding, Multimeric Structure Formation, and Immunogenicity of Viral Capsid Proteins

Viral capsid proteins are widely explored for subunit vaccine development but are often hampered by their complexity of production and low immunogenicity. Here, we report a simple approach to overcome these challenges by combining mRNA vaccine technology with protein engineering. Using African swine fever virus (ASFV) capsid proteins P72 and penton as models, we engineered them into membrane-bound and secreted forms, and compared their immunogenicity to the native intracellular form in mice and pigs through mRNA vaccination. The membrane-bound and secreted P72 and penton folded into their native multimeric structure independent of viral chaperone, therefore preserving their conformational epitopes. The membrane-bound P72 and penton also elicited significantly stronger antibody and T cell responses than their secreted or intracellular counterparts. Our study provides a simple approach to enhance folding, multimeric structure formation, and immunogenicity of viral capsid proteins for ASFV subunit vaccine development and immunogenicity of intracellular proteins in general.

immunology↗