Cellular Responses Research Articles

Cellular immune response against rabies vaccination in laboratory-bred dogs: A systematised review.

The immunogenicity of rabies vaccines is commonly measured by serological testing, which includes measuring rabies virus-neutralising antibody titre levels in the serum. Apart from humoral immunity, cellular immunity measurements are also helpful in assessing the immunogenicity and efficacy of rabies vaccinations. Recently, there has been an increased emphasis on cellular immunity measurements against rabies in humans and animals. This review aimed to systematically analyse the literature on the composition of cellular immune responses against rabies vaccination in laboratory dogs. A literature survey was conducted to collect suitable articles by searching the research databases PubMed, Web of Science and Scopus. Subsequently, a two-person screening was conducted to identify suitable articles based on predetermined inclusion and exclusion criteria. Data extraction was performed by two authors independently. A total of 1396 studies were identified from the initial search. Following the screening, six studies were selected for final review. Different methods of detecting immune parameters from peripheral blood mononuclear cells were identified from the studies. Reports have demonstrated positive outcomes of the use of adjuvants for cellular immunity development. Even though the lack of specific immunological techniques and the specific reagents have negatively affected these types of cellular immunity measurements, it was evident that combining both humoral and cellular immune parameters against the rabies antigen would provide a clearer picture of the level of responsiveness in animals towards vaccinations and the protection against the disease. The protocol for this review was published in the International Prospective Register of Systematic Review (CRD42022380023).

Comparison of immune responses to respiratory syncytial virus in infancy, childhood, and adulthood using an in vitro model of human respiratory infection.

Respiratory syncytial virus (RSV) is a major contributor to morbidity and mortality in infants. We developed an in vitro model of human respiratory infection to study cellular immune responses to RSV in infants, children, and adults. The model includes human lung epithelial A549 cells or human fetal lung fibroblasts infected with a clinical strain of RSV at a multiplicity of infection of 0.3, cocultured with human cord blood mononuclear cells (CBMCs) or peripheral blood mononuclear cells (PBMCs). Mononuclear cells were collected at multiple ages ranging from birth to adulthood. After 20 h of incubation, flow cytometry was used to measure CBMC/PBMC responses to RSV. A549s were more permissive to RSV and when infected produced more CCL5, CCL11, and CXCL9; less CSF-3, CXCL10, interleukin (IL)-1α, IL-1RA, and IL-6; and similar CCL2, CCL3, CCL4, CCL7, CXCL1, CXCL11, IL-1β, IL-7, IL-8, and tumor necrosis factor α compared with fibroblasts; A594s were used for subsequent experiments. CBMCs/PBMCs upregulated multiple markers of activation, maturation, and degranulation upon exposure to RSV-infected A549s. Interferon γ expression in natural killer, CD4, and CD8 cells and CD107a expression in natural killer cells showed a gradual increase from infancy to adulthood. IL-12 expression in dendritic cells and monocytes was highest in adult PBMCs. Our in vitro model of human RSV infection recapitulated the expected bias away from T helper 1 and effector responses to RSV infection in infancy and revealed changes in innate and adaptive RSV-specific cellular immune responses over time.

Curcumin enhances the anti-obesogenic activity of orlistat through SKN-1/NRF2-dependent regulation of nutrient metabolism in Caenorhabditis elegans.

Metabolic dysregulation, a defining feature of obesity, disrupts essential signalling pathways involved in nutrient sensing and mitochondria homeostasis. The nuclear factor erythroid 2-related factor 2 (NRF-2) serves as a pivotal regulator of the cellular stress response, and recent studies have implicated it in the pathogenesis of obesity, diabetes, and metabolic syndrome. Curcumin, a polyphenolic compound derived from turmeric, has been identified as a potent activator of NRF-2. Evidence suggests curcumin impacts obesity and metabolic disorders by modulating gut microbiota composition, increasing energy expenditure, and regulating lipid metabolism. Orlistat, an anti-obesity drug, inhibits fat absorption in the gastrointestinal tract, but its side effects limits its broader use. The present study aims to investigate the potential synergetic effect of a hybrid combination between orlistat and curcumin. Additionally, we provide a detailed understanding of the molecular mechanisms through which this combination mitigates glucose-induced lipid accumulation in Caenorhabditis elegans, with a focus on the role of the skinhead 1 (SKN-1) transcription factor, an orthologue of NRF2. We assessed the lipid accumulation and the changes in skn-1 transcriptional activity in C. elegans using confocal GFP-based detection, alongside mRNA expression analysis of genes from lipid metabolism and oxidative stress response in wild-type, QV225 and LD1 strains. Furthermore, we evaluated locomotion, chemotaxis and mitochondrial dynamics to enhance our understanding of the proposed molecular-based model. Our findings reveal that the orlistat/curcumin combination exerts an anti-obesogenic effect through SKN-1/NRF2-dependent regulation of conserved genes involved in carbohydrate and lipid metabolism in C. elegans. Moreover, the combination stimulates mitochondrial potential, further contributing to the observed synergistic effects. The hybrid combination of orlistat and curcumin demonstrates significant anti-obesity activity by regulating nutrient-sensing pathways through SKN-1/NRF-2 modulation. This approach may allow for the reduction of orlistat dosage, thereby minimizing its adverse effects while maintaining its therapeutic efficacy.

Mixed lipopeptide-based mucosal vaccine candidate induces cross-variant immunity and protects against SARS-CoV-2 infection in hamsters.

The global dissemination of SARS-CoV-2 led to a worldwide pandemic in March 2020. Even after the official downgrading of the COVID-19 pandemic, infection with SARS-CoV-2 variants continues. The rapid development and deployment of SARS-CoV-2 vaccines helped to mitigate the pandemic to a great extent. However, the current vaccines are suboptimal; they elicit incomplete and short-lived protection and are ineffective against evolving virus variants. Updating the spike antigen according to the prevailing variant and repeated boosters is not the long-term solution. We have designed a lipopeptide-based, mucosal, pan-coronavirus vaccine candidate, derived from highly conserved and/or functional regions of the SARS-CoV-2 spike, nucleocapsid, and membrane proteins. Our studies demonstrate that the designed lipopeptides (LPMix) induced both cellular and humoral (mucosal and systemic) immune responses upon intranasal immunization in mice. Furthermore, the antibodies bound to the wild-type and mutated S proteins of SARS-CoV-2 variants of concern, including Alpha, Beta, Delta and Omicron, and also led to efficient neutralization in a surrogate viral neutralization assay. Our sequence alignment and 3-dimensional molecular modeling studies demonstrated that spike-derived epitopes, P1 and P2, are sequentially and/or structurally conserved among the SARS-CoV-2 variants. The addition of a novel mucosal adjuvant, heat-killed Caulobacter crescentus, to the lipopeptide vaccine significantly bolstered mucosal antibody responses. Finally, the lipopeptide-based intranasal vaccine demonstrated significant improvement in lung pathologies in a hamster model of SARS-CoV-2 infection. These studies are fundamentally important and open new avenues in the investigation of an innovative, broadly protective intranasal vaccine platform for SARS-CoV-2 and its variants.

High-intensity focused ultrasound ablation to increase tumor-specific lymphocytes in prostate cancer.

Treatment options for localized prostate cancer have been expanded by FDA-approval of High-Intensity Focused Ultrasound (HIFU). Prostate cancer typically has few tumor-infiltrating lymphocytes, which are crucial for antitumor immunity. This study investigated the use of HIFU to increase lymphocyte infiltration into the tumor and enhance antitumor immunity. RM1 prostate tumors were implanted onto both flanks of syngeneic C57BL/6 J mice, with one tumor subjected to HIFU treatment. The growth of the contralateral tumor was monitored. Blood samples were obtained from patients both before and after prostatectomy or HIFU treatment. Peripheral blood mononuclear cells (PBMCs) were then isolated to analyze the immune cells. In murine experiments, the application of HIFU to one tumor decreased the growth of the contralateral (non-HIFU treated) tumor, when the contralateral tumor was the same tumor type, but not when it was a different tumor type. HIFU increased infiltration of CD4+ and CD8+ lymphocytes into the contralateral, same-type tumor. Lymphocyte depletion studies affirmed that the antitumor immune response triggered by HIFU relies on CD4+ and CD8+ lymphocytes. Addition of cholesterol-lowering intervention further increased antitumor immunity generated by HIFU in mice. In human subjects, HIFU, but not prostatectomy, stimulated anti-tumor CD4+ and CD8+ lymphocytes. We concluded that HIFU induced a potent cellular antitumor immune response that inhibited the progression of murine prostate tumors. HIFU stimulated tumor-specific cellular immunity in patients. Future clinical trials should explore the clinical benefits of HIFU, possibly in combination with existing immunotherapies, as immune modulators for both localized and metastatic disease.

The oxygen level in air directs airway epithelial cell differentiation by controlling mitochondrial citrate export.

Oxygen controls most metazoan metabolism, yet in mammals, tissue O2 levels vary widely. While extensive research has explored cellular responses to hypoxia, understanding how cells respond to physiologically high O2 levels remains uncertain. To address this problem, we investigated respiratory epithelia as their contact with air exposes them to some of the highest O2 levels in the body. We asked how the O2 level in air controls differentiation of airway basal stem cells into the ciliated epithelial cells essential for clearing airborne pathogens from the lung. Through a metabolomics screen and 13C tracing on primary cultures of human airway basal cells, we found that the O2 level in air directs ciliated cell differentiation by increasing mitochondrial citrate export. Unexpectedly, disrupting mitochondrial citrate export elicited hypoxia transcriptional responses independently of HIF1α stabilization and at O2 levels that would be hyperoxic for most tissues. These findings identify mitochondrial citrate export as a cellular mechanism for responding to physiologically high O2 levels.

Transcriptome-wide alternative mRNA splicing analysis reveals post-transcriptional regulation of neuronal differentiation.

Alternative splicing (AS) plays an important role in neuronal development, function, and disease. Efforts to analyze the transcriptome of AS inneurons on a wide scale are currently limited. We characterized the transcriptome-wide AS changes in SH-SY5Y neuronal differentiation model, which is widely used to study neuronal function and disorders. Our analysis revealed global changes in five AS programs that drive neuronal differentiation. Motif analysis revealed the contribution of RNA-binding proteins (RBPs) to the regulation of AS during neuronal development. We concentrated on the primary alternative splicing program that occurs during differentiation, specifically on events involving exon skipping (SE). Motif analysis revealed motifs for polypyrimidine tract-binding protein 1 (PTB) and ELAV-like RNA binding protein 1 (HuR/ELAVL1) to be the top enriched in SE events, and their protein levels were downregulated after differentiation. shRNA knockdown of either PTB and HuR was associated with enhanced neuronal differentiation and transcriptome-wide exon skipping events that drive the process of differentiation. At the level of gene expression, we observed only modest changes, indicating predominant post-transcriptional effects of PTB and HuR. We also observed that both RBPs altered cellular responses to oxidative stress, in line with the differentiated phenotype observed after either gene knockdown. Our work characterizes the AS changes in a widely used and important model of neuronal development and neuroscience research and reveals intricate post-transcriptional regulation of neuronal differentiation.

Gut Microbiota-Bone Axis.

Knowledge of the complex interplay between gut microbiota and human health is gradually increasing as it has just recently been a field of such great interest. Recent studies have reported that communities of microorganisms inhabiting the gut influence the immune system through cellular responses and shape many physiological and pathophysiological aspects of the body, including muscle and bone metabolism (formation and resorption). Specifically, the gut microbiota affects skeletal homeostasis through changes in host metabolism, the immune system, hormone secretion, and the gut-brain axis. The major role on gut-bone axis is due to short-chain fatty acids (SCFAs). They have the ability to influence regulatory T-cell (Tregs) development and activate bone metabolism through the action of Wnt10. SCFA production may be a mechanism by which the microbial community, by increasing the serum level of insulin-like growth factor 1 (IGF-1), leads to the growth and regulation of bone homeostasis. A specific SCFA, butyrate, diffuses into the bone marrow where it expands Tregs. The Tregs induce production of the Wnt ligand Wnt10b by CD8+ T cells, leading to activation of Wnt signaling and stimulation of bone formation. At the hormonal level, the effect of the gut microbiota on bone homeostasis is expressed through the biphasic action of serotonin. Some microbiota, such as spore-forming microbes, regulate the level of serotonin in the gut, serum, and feces. Another group of bacterial species (Lactococcus, Mucispirillum, Lactobacillus, and Bifidobacterium) can increase the level of peripheral/vascular leptin, which in turn manages bone homeostasis through the action of brain serotonin.

Comprehensive quantification of C4 to C26 free fatty acids using a supercritical fluid chromatography-mass spectrometry method in pharmaceutical-grade egg yolk powders intended for total parenteral nutrition use.

Free fatty acids (FFAs) are important energy sources and significant for energy transport in the body. They also play a crucial role in cellular oxidative stress responses, following cell membrane depolarization, making accurate quantification of FFAs essential. This study presents a novel supercritical fluid chromatography-mass spectrometry (SFC-MS) method using selected ion recording in negative electrospray ionization mode, enabling rapid quantification of 31 FFAs within 6min without derivatization. FFAs are identified and quantified using an HSS C18 SB column and a secondary mobile phase consisting of methanol with formic acid by detecting their [M - H]- ions. Calibration curves showed strong linearity (R2 ≥ 0.9910), spanning 1000-12,000ng/mL for short-chain FFAs and 50-1200ng/mL for medium- and long-chain FFAs. The method achieves detection limits as low as 1ng/µL for short-chain FFAs and 0.05pg/µL for other FFAs per on-column injection. The method demonstrated high accuracy and precision, with bias and coefficients of variation maintained below 15% across five quality control levels. Freeze-thaw and autosampler stability studies confirmed the behavior of matrix-matched standards under optimal storage conditions. The validated method was applied to the analysis of pharmaceutical-grade egg yolk powders, using 13 deuterated FFAs as internal standards (IS) in comparison with heptadecanoic acid (C17:0). Significant variations in FFA quantification using two different IS approaches underscore the importance of selecting an appropriate IS. In summary, this study introduces a reliable and validated SFC-MS method for analyzing FFAs ranging from C4 to C26, requiring minimal sample preparation.

Inhibition of vascular smooth muscle cell PERK/ATF4 ER stress signaling protects against abdominal aortic aneurysms.

Abdominal aortic aneurysms (AAA) are a life-threatening cardiovascular disease for which there is a lack of effective therapy preventing aortic rupture. During AAA formation, pathological vascular remodeling is driven by vascular smooth muscle cell (VSMC) dysfunction and apoptosis, for which the mechanisms regulating loss of VSMCs within the aortic wall remain poorly defined. Using single-cell RNA-Seq of human AAA tissues, we identified increased activation of the endoplasmic reticulum stress response pathway, PERK/eIF2α/ATF4, in aortic VSMCs resulting in upregulation of an apoptotic cellular response. Mechanistically, we reported that aberrant TNF-α activity within the aortic wall induces VSMC ATF4 activation through the PERK endoplasmic reticulum stress response, resulting in progressive apoptosis. In vivo targeted inhibition of the PERK pathway, with VSMC-specific genetic depletion (Eif2ak3fl/fl Myh11-CreERT2) or pharmacological inhibition in the elastase and angiotensin II-induced AAA model preserved VSMC function, decreased elastin fragmentation, attenuated VSMC apoptosis, and markedly reduced AAA expansion. Together, our findings suggest that cell-specific pharmacologic therapy targeting the PERK/eIF2α/ATF4 pathway in VSMCs may be an effective intervention to prevent AAA expansion.

Development of a novel multi-epitope mRNA vaccine candidate to combat SFTSV pandemic.

Severe Fever with Thrombocytopenia Syndrome virus (SFTSV) is a novel identified pathogen, despite two decades of research on SFTSV, the potential widespread threats pose a significant challenge for researchers in developing new treatment and prevention methods. In this present, we have developed a multi-epitope mRNA vaccine for SFTSV and valid it with in silico methods. We screened 9 immunodominant epitopes for cytotoxic T cells (CTL), 7 for helper T cells (HTL), and 8 for Linear B-cell (LBL) based on promising candidate protein Gn, Gc, Np, and NSs. All predicted epitopes demonstrated strong antigenicity without any potential harm to humans. Additionally, the high conservancy is required to cover different strains. All epitopes as well as adjuvants were constructed into a final vaccine, which was further assesd by calculating of physicochemical properties. Then, we docked the vaccine protein with immune receptors and analyzed the complexes with dynamic simulations to evaluate its affinity to receptors. Finally, the vaccine sequence was constructed into a mRNA sequence. The constructed vaccine is a potential candidate for combating SFTSV by stimulating protective humoral and cellular immune responses.

Mitochondrial protein import stress.

Mitochondria have to import a large number of precursor proteins from the cytosol. Chaperones keep these proteins in a largely unfolded state and guide them to the mitochondrial import sites. Premature folding, mitochondrial stress and import defects can cause clogging of import sites and accumulation of non-imported precursors, representing a critical burden for cellular proteostasis. Here we discuss how cells respond to mitochondrial protein import stress by regenerating clogged import sites and inducing stress responses. The mitochondrial protein import machinery has a dual role by serving as sensor for detecting mitochondrial dysfunction and inducing stress-response pathways. The production of chaperones that fold or sequester precursor proteins in deposits is induced and the proteasomal activity is increased to remove the excess precursor proteins. Together, these pathways reveal how mitochondria are tightly integrated into a cellular proteostasis and stress response network to maintain cell viability.

Clinical utility of the lymphocyte proliferation assay, an in vitro functional readout of the adaptive immune response.

Despite great advancements in the discovery of genetic variants underlying inborn errors of immunity, functional assessment of the immunological profile of patients in routine clinical practice remains challenging. The lymphocyte proliferation assay using 3H-thymidine incorporation has been the gold standard for decades for functional evaluation of T cells in the clinical laboratory, however, recently developed flow cytometry methods allowing for single cell analysis provide non-radioactive alternatives. Understanding the technical and analytical challenges of test development, validation and maintenance is essential for correct interpretation and test utilization, to assure appropriate and timely patient care. This review will discuss the technological aspects, validation and clinical utilization of in vitro lymphocyte proliferation assays performed in the clinical immunology laboratory, providing a diagnostic readout for T lymphocyte function, an essential hallmark of a functional cellular immune response, and allowing for the detection of impaired responses such as in patients with functional T cell defects.

Composite Graphene for the Dimension- and Pore-Size-Mediated Stem Cell Differentiation to Bone Regenerative Medicine.

As one of the most promising means to repair diseased tissues, stem cell therapy with immense potential to differentiate into mature specialized cells has been rapidly developed. However, the clinical application of stem-cell-dominated regenerative medicine was heavily hindered by the loss of pluripotency during the long-term in vitro expansion. Here, a composite three-dimensional (3D) graphene-based biomaterial, denoted as GO-Por-CMP@CaP, with hierarchical pore structure (micro- to macropore), was developed to guide the directional differentiation of human umbilical cord MSCs (hucMSCs) into osteoblasts. GO-Por-CMP@CaP could act as a high-efficiency living composite material without a "dead space", effectively regulating the cellular response. The 3D topological structure generated via the two-step modification on two-dimensional graphene could effectively mimic the natural 3D microenvironment of cells, enhancing the stem cell attachment, which is not only conducive for the proliferation of stem cells but also beneficial for the osteogenic differentiation. Meanwhile, the wide existence of interconnected macropores was favorable for bone ingrowth, capillary formation, as well as the nutrients transportation. Furthermore, the concurrent existence of micro- and mesopores significantly promoted the extracellular matrix (ECM) adsorption, which ensured cellular attachment, leading to multiscale osteointegration. Both in vitro and in vivo assay demonstrated the above three factors collaborated mutually with nanosized calcium phosphate (CaP, with chemical similarities to the inorganic components of bone), which provided abundant adhesive sites to adequately induce osteogenic differentiation in the absence of any soluble growth factors. Proteomic analysis experiments confirmed that GO-Por-CMP@CaP promoted the differentiation of hucMSCs cells into osteoblasts by affecting the PI3K-Akt signaling pathway through the up-regulation of SPP1 protein. Our study offers a pure material-based stem cell differentiation regulating behavior via engineering the dimension and porosity of material, which provides insights into the design and development of substitutes to bone repair materials.

Green revolution gene drives adaptation of Arabidopsis to the extremely high altitude.

To elucidate the process of adaptation, particularly the traits subject to natural selection and the molecular mechanisms underlying their natural variation, is one of the primary objectives of evolutionary biology. The uplifted landscape offers an excellent framework for understanding how organisms adapt to dramatic climatic gradients. To investigate the genetic basis of plant adaptation to the extremely high altitude, we first compared the genomic and phenotypic variations of two closely related Arabidopsis thaliana accessions from high altitude (Xizang, also known as "Tibet") and low altitude (Yunnan), respectively. The Xizang population represents a relict group characterized by a small effective population size. Notably, the Xizang genome has more transposable elements (TEs) and more gene loss-of-function (LoF) mutations. Differentially expressed genes were enriched in biological processes of cellular response to oxygen-containing compound, regulation of defense response, and response to light intensity. Intriguingly, the phenotypic selection analysis revealed that silique density was under natural selection. Furthermore, we genetically mapped and validated that the LoF mutation of GA20ox1, the homologous gene of green revolution in rice, resulted in a higher silique density in Xizang Arabidopsis. Given that GA20ox1 is linked to Arabidopsis adaptation to the Alps Mountains, its parallel evolution plays an important role in the adaptation to Alpine habitats. Overall, our results highlight that identifying adaptive traits and elucidating the molecular mechanisms underlying natural variation of these traits is crucial for unraveling the mystery of adaptive evolution and has significant implications for crop breeding.

A comprehensive review of arsenic-induced neurotoxicity: Exploring the role of glial cell pathways and mechanisms.

The review aims to examine the neurotoxic effects of arsenic, particularly exploring the roles of glial cells-astrocytes, microglia, and oligodendrocytes, amid its widespread environmental contamination and impact on cognitive impairments. It highlights the role of altered neurotrophin and growth factor signaling in disrupting neuronal health and cognitive performance. It elucidates the intricate interactions between oxidative stress, DNA damage, neurotransmitter disruption, and cellular signaling alterations, underscoring the vital importance of the glial cells. These cells are crucial for preserving neural health and responding to environmental toxins, and arsenic disrupts their functions, resulting in decreased antioxidative responses, induction of inflammatory pathways, and subsequent neuronal dysfunction. The brain's cytotoxic impact arises from a complex network of cellular responses, with pathways such as MAPK, transcription factor and autophagy signaling to play critical roles in mediating these dysregulated inflammation and oxidative stress mechanisms. The detailed exploration into specific impacts of arsenic on glial cell morphology, activation, and mitochondrial functions illuminates the cascade of neuroinflammatory and neurodegenerative changes that may be triggered upon arsenic exposure. The review recommends a multidisciplinary research approach by emphasizing the significance of the brain's microenvironment, methylation processes, and the enzyme AS3MT in arsenic neurotoxicity. It calls for converging environmental science, neurobiology, and toxicology to develop targeted interventions for preventing and mitigating arsenic's neurotoxic effects. This in-depth exploration into glial cell dynamics aims to advance public health and neurotoxicology research, striving to devise strategies that reduce the cognitive and neurodegenerative damage caused by arsenic, thereby enhancing global health outcomes.

Short-term oral exposure to nanoplastics does not significantly impact the antiviral immune response of the mouse.

The increasing prevalence of nanoplastics (NPs) in the environment, particularly polystyrene (PS) nanoparticles, raises concerns regarding their potential impact on human and animal health. Given their small size, NPs can cross biological barriers and accumulate in organs, including those critical for immune functions. This study investigates the effects of short-term oral exposure to 100 and 500 nm PS NPs on the adaptive immune responses during viral infections in vivo, using vesicular stomatitis virus (VSV) and lymphocytic choriomeningitis virus (LCMV) as models. Male and female C57BL/6 mice were orally exposed to PS NP for a period of 28 days, during which they were infected with either VSV or LCMV to study the humoral and cellular responses, respectively. The humoral responses were assessed by measuring total and VSV-specific antibody levels, and splenic immune populations. T cell phenotypes, activation, exhaustion and functionality towards LCMV epitopes were studied as readouts of the cellular responses. Our results demonstrate that short-term NP exposure does not significantly affect the generation or neutralizing capacity of antibodies against VSV, nor the cellular responses directed against LCMV. These findings indicate that, under these conditions, PS NP exposure does not significantly compromise the adaptive immune responses during viral infections, underscoring the value of in vivo models.

A Universal Therapeutic Vaccine Leveraging Autologous Pre-Existing Immunity to Eliminate in Situ Uniformly Engineered Heterogeneous Tumor Cells.

Tumor vaccines that activate the autologous immune system to eliminate tumor cells represent a promising approach in cancer immunotherapy. However, challenges such as tumor heterogeneity, limited antigen selection, insufficient antigen presentation, and the slow onset of de novo immune responses have resulted in poor universality and suboptimal response rates. In contrast, pathogen-specific pre-existing immunity acquired through infection or vaccination, can rapidly generate a more potent and enduring immune response upon re-encounter with the same antigen. Here, an adeno-associated virus (AAV)-based therapeutic vaccine capable of genetically modifying diverse tumor cells to uniformly overexpress and efficiently present the highly immunogenic transmembrane SARS-CoV-2 receptor binding domain (RBD), and to release RBD-enveloped virus-like particles, which awaken and enhance the RBD-specific pre-existing immunity, leading to significant tumor remission, is engineered. Mechanistically, this therapeutic vaccine leverages the robust RBD-specific pre-existing immunity and heightened antibody-mediated phagocytosis to eliminate engineered tumor cells while inducing antigen spreading, thereby provoking a more diverse tumor-specific cellular immune responses. Notably, widespread administration of vaccines against various pathogens has provided a versatile pool of pre-existing immunity that can be redirected to eradicate tumors. These findings offer a novel perspective on overcoming the limitations posed by tumor heterogeneity and personalized medicine.

De novo genome assembly and annotations of Bombus lapidarius and Bombus niveatus provide insights into the environmental adaptability

Bumblebees are ubiquitous, cold-adapted, primitively eusocial bees and important pollinators for crops and vegetation. However, many species are declining worldwide due to multiple factors, including human-induced habitat loss, agricultural chemicals, global warming, and climate change. In particular, future climate scenarios predict a shift in the spatial distribution of bumblebees under global warming, with some species declining and others potentially expanding. Here, we report a de novo genome assembly and annotation for Bombus lapidarius and Bombus niveatus to decipher species-specific potential genomic capacity against such environmental stressors. With harboring more than 23,000 protein-coding genes, the assembled genomes of B. lapidarius and B. niveatus are 244.44 Mb (scaffold N50 of 9.45 Mb) and 259.84 Mb (scaffold N50 of 10.94 Mb), respectively, which exhibit similar trends in terms of genome size and composition with other bumblebees. Gene family analysis reveals differences in species-specific expanded gene families. B. lapidarius exhibits expanded genes related to pre/postsynaptic organization, while B. niveatus shows a distinct expansion in gene families regulating cellular growth, aging, and responses to abiotic and biotic stressors, such as those containing SCAN domains, WD-repeats, and Ras-related proteins. Our genome-wide screens revealed positive selection on environmental stress-responsive genes such as dip2, yme1l, and spg7 in B. lapidarius, whereas positive selection signatures were found in genes such as myd88, mybbp1A, and rhau, which are involved in environmental stress resistance for B. niveatus. These high-quality genome assemblies and comparative genome analysis unveil potential drivers that underlie genome evolution in bumblebees, offering valuable insights into environmental adaptation and conservation efforts.

Incomplete elimination of viral genomes is associated with chronic inflammation in nonhuman primate livers after AAV-mediated gene transfer.

The liver is a unique organ where immunity can be biased toward ineffective response notably in the context of viral infections. Chronic viral hepatitis depends on the inability of the T-cell immune response to eradicate antigen. In the case of recombinant Adeno-Associated-Virus, used for therapeutic gene transfer, conflicting reports describe tolerance induction to different transgene products while other studies have shown conventional cytotoxic CD8+ T cell responses with a rapid loss of transgene expression. We performed a 1 year follow up of 6 non-human primates after all animals received an rAAV8 vector carrying the GFP transgene at doses of 7×1012 vg/kg. We report that despite anti-GFP peripheral cellular response and loss of hepatic transgene expression, we were still able to detect persisting viral genomes in the liver until 1-year post-injection. These viral genomes were associated with liver inflammation, fibrosis and signs of CD8 T cell exhaustion, including high expression of PD-1. Our study shows that AAV8-mediated gene transfer can results to loss of transgene expression in liver and chronic inflammation several months after gene transfer.