A TLR4-Targeting Bioactive Peptide Hydrogel to Regulate Immune-Microenvironment for Diabetic Wound Repair.

Persistent inflammation and disrupted immunoregulation are critical factors in impeding diabetic wound healing. While immunoregulatory hydrogel dressings hold significant promise for clinical applications in diabetic wound healing, the current application often demands intricate interventions and high-cost treatments involving cytokines and cell therapies. The development of single component immunoregulatory hydrogels remains a complex challenge. To address this issue, an active peptide hydrogel with immunoregulatory properties targeting the TLR4/NF-kB pathway, aiming to promote rapid diabetic wound healing, is engineered. The hydrogel sequence comprises naphthalene derivative, phenylalanine, and glycine to modulate hydrophilic/hydrophobic characteristics. The amino group on arginine contributes to tissue adhesion and regulation of intermolecular forces, ultimately yielding stable gels. The results underscore the formation of the peptide hydrogel (NFA) via the physical crosslinking of self-assembled nanofibers in water, thereby affording both excellent injectability and tissue adhesion. Notably, NFA demonstrates significant potential in promoting wound healing in a mouse model with full-thickness wounds by regulating macrophage responses in the inflammatory microenvironment through the TLR4/NF-kB pathway.

A therapeutic hepatitis B mRNA vaccine with strong immunogenicity and persistent virological suppression.

Here we report on the development and comprehensive evaluations of an mRNA vaccine for chronic hepatitis B (CHB) treatment. In two different HBV carrier mouse models generated by viral vector-mediated HBV transfection (pAAV-HBV1.2 and rAAV8-HBV1.3), this vaccine demonstrates sufficient and persistent virological suppression, and robust immunogenicity in terms of induction of strong innate immune activation, high-level virus-specific antibodies, memory B cells and T cells. mRNA platform therefore holds prospects for therapeutic vaccine development to combat CHB.

Herpes zoster mRNA vaccine induces superior vaccine immunity over licensed vaccine in mice and rhesus macaques.

Herpes zoster remains an important global health issue and mainly occurs in aged and immunocompromised individuals with an early exposure history to Varicella Zoster Virus (VZV). Although the licensed vaccine Shingrix has remarkably high efficacy, undesired reactogenicity and increasing global demand causing vaccine shortage urged the development of improved or novel VZV vaccines. In this study, we developed a novel VZV mRNA vaccine candidate (named as ZOSAL) containing sequence-optimized mRNAs encoding full-length glycoprotein E encapsulated in an ionizable lipid nanoparticle. In mice and rhesus macaques, ZOSAL demonstrated superior immunogenicity and safety in multiple aspects over Shingrix, especially in the induction of strong T-cell immunity. Transcriptomic analysis revealed that both ZOSAL and Shingrix could robustly activate innate immune compartments, especially Type-I IFN signalling and antigen processing/presentation. Multivariate correlation analysis further identified several early factors of innate compartments that can predict the magnitude of T-cell responses, which further increased our understanding of the mode of action of two different VZV vaccine modalities. Collectively, our data demonstrated the superiority of VZV mRNA vaccine over licensed subunit vaccine. The mRNA platform therefore holds prospects for further investigations in next-generation VZV vaccine development.

Multivalent antigen display on nanoparticle immunogens increases B cell clonotype diversity and neutralization breadth to pneumoviruses.

Nanoparticles for multivalent display and delivery of vaccine antigens have emerged as a promising avenue for enhancing B cell responses to protein subunit vaccines. Here, we evaluated B cell responses in rhesus macaques immunized with prefusion-stabilized respiratory syncytial virus (RSV) F glycoprotein trimer compared with nanoparticles displaying 10 or 20 copies of the same antigen. We show that multivalent display skews antibody specificities and drives epitope-focusing of responding B cells. Antibody cloning and repertoire sequencing revealed that focusing was driven by the expansion of clonally distinct B cells through recruitment of diverse precursors. We identified two antibody lineages that developed either ultrapotent neutralization or pneumovirus cross-neutralization from precursor B cells with low initial affinity for the RSV-F immunogen. This suggests that increased avidity by multivalent display facilitates the activation and recruitment of these cells. Diversification of the B cell response by multivalent nanoparticle immunogens has broad implications for vaccine design.

Performance Evaluation for Clinical Stroke Rehabilitation via an Automatic Mobile Gait Trainer.

This paper investigates the clinical efficacy of an automatic mobile trainer for gait training in stroke patients. Neuro-Developmental Treatment (NDT) is a rehabilitation method for stroke patients that enhances motor learning through repeated practice. Despite the proven effectiveness of therapist-assisted NDT, it is labor-intensive and demands health resources. Therefore, we developed automatic trainers based on NDT principles to perform gait training. This paper modifies the mobile trainer's intervention patterns to improve the subject's longitudinal gait symmetry, lateral pelvic displacement symmetry, and pelvic rotation. We first invited ten healthy subjects to test the modified trainer and then recruited 26 stroke patients to undergo the same gait training. Longitudinal symmetry, lateral symmetry, and pelvic rotation were assessed before, during, and after the intervention. Most subjects show improvements in longitudinal symmetry, lateral symmetry, and pelvic rotation after using the trainer. These results confirm the trainer's effectiveness of the modified intervention schemes in helping clinical gait rehabilitation for stroke patients.

Unmodified rabies mRNA vaccine elicits high cross-neutralizing antibody titers and diverse B cell memory responses.

Licensed rabies virus vaccines based on whole inactivated virus are effective in humans. However, there is a lack of detailed investigations of the elicited immune response, and whether responses can be improved using novel vaccine platforms. Here we show that two doses of a lipid nanoparticle-formulated unmodified mRNA vaccine encoding the rabies virus glycoprotein (RABV-G) induces higher levels of RABV-G specific plasmablasts and T cells in blood, and plasma cells in the bone marrow compared to two doses of Rabipur in non-human primates. The mRNA vaccine also generates higher RABV-G binding and neutralizing antibody titers than Rabipur, while the degree of somatic hypermutation and clonal diversity of the response are similar for the two vaccines. The higher overall antibody titers induced by the mRNA vaccine translates into improved cross-neutralization of related lyssavirus strains, suggesting that this platform has potential for the development of a broadly protective vaccine against these viruses.

Algorithm for optimized mRNA design improves stability and immunogenicity.

Messenger RNA (mRNA) vaccines are being used to combat the spread of COVID-19 (refs. 1-3), but they still exhibit critical limitations caused by mRNA instability and degradation, which are major obstacles for the storage, distribution and efficacy of the vaccine products4. Increasing secondary structure lengthens mRNA half-life, which, together with optimal codons, improves protein expression5. Therefore, a principled mRNA design algorithm must optimize both structural stability and codon usage. However, owing to synonymous codons, the mRNA design space is prohibitively large-for example, there are around 2.4 × 10632 candidate mRNA sequences for the SARS-CoV-2 spike protein. This poses insurmountable computational challenges. Here we provide a simple and unexpected solution using the classical concept of lattice parsing in computational linguistics, where finding the optimal mRNA sequence is analogous to identifying the most likely sentence among similar-sounding alternatives6. Our algorithm LinearDesign finds an optimal mRNA design for the spike protein in just 11 minutes, and can concurrently optimize stability and codon usage. LinearDesign substantially improves mRNA half-life and protein expression, and profoundly increases antibody titre by up to 128 times in mice compared to the codon-optimization benchmark on mRNA vaccines for COVID-19 and varicella-zoster virus. This result reveals the great potential of principled mRNA design and enables the exploration of previously unreachable but highly stable and efficient designs. Our work is a timely tool for vaccines and other mRNA-based medicines encoding therapeutic proteins such as monoclonal antibodies and anti-cancer drugs7,8.

Delayed generation of functional virus-specific circulating T follicular helper cells correlates with severe COVID-19.

Effective humoral immune responses require well-orchestrated B and T follicular helper (Tfh) cell interactions. Whether these interactions are impaired and associated with COVID-19 disease severity is unclear. Here, longitudinal blood samples across COVID-19 disease severity are analysed. We find that during acute infection SARS-CoV-2-specific circulating Tfh (cTfh) cells expand with disease severity. SARS-CoV-2-specific cTfh cell frequencies correlate with plasmablast frequencies and SARS-CoV-2 antibody titers, avidity and neutralization. Furthermore, cTfh cells but not other memory CD4 T cells, from severe patients better induce plasmablast differentiation and antibody production compared to cTfh cells from mild patients. However, virus-specific cTfh cell development is delayed in patients that display or later develop severe disease compared to those with mild disease, which correlates with delayed induction of high-avidity neutralizing antibodies. Our study suggests that impaired generation of functional virus-specific cTfh cells delays high-quality antibody production at an early stage, potentially enabling progression to severe disease.

Single-cell transcriptome atlas reveals protective characteristics of COVID-19 mRNA vaccine.

Messenger RNA (mRNA) vaccines are promising alternatives to conventional vaccines in many aspects. We previously developed a lipopolyplex (LPP)-based mRNA vaccine (SW0123) that demonstrated robust immunogenicity and strong protective capacity against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection in mice and rhesus macaques. However, the immune profiles and mechanisms of pulmonary protection induced by SW0123 remain unclear. Through high-resolution single-cell analysis, we found that SW0123 vaccination effectively suppressed SARS-CoV-2-induced inflammatory responses by inhibiting the recruitment of proinflammatory macrophages and increasing the frequency of polymorphonuclear myeloid-derived suppressor cells. In addition, the apoptotic process in both lung epithelial and endothelial cells was significantly inhibited, which was proposed to be one major mechanism contributing to vaccine-induced lung protection. Cell-cell interaction in the lung compartment was also altered by vaccination. These data collectively unravel the mechanisms by which the SW0123 protects against lung damage caused by SARS-CoV-2 infection.

Superior Gait Symmetry and Postural Stability among Yoga Instructors-Inertial Measurement Unit-Based Evaluation.

This study investigates gait symmetry and single-leg stance balance of professional yoga instructors versus age-matched typically developed controls using inertial measurement unit (IMU)-based evaluation. We recruited twenty-five yoga instructors and twenty-five healthy control subjects to conduct the walking experiments and single-leg stance tests. Kinematic data were measured by attaching IMUs to the lower limbs and trunk. We assessed the asymmetry of swing phases during the normal-walk and tandem-walk tests with eyes open and closed, respectively. The subjects subsequently conducted four single-leg stance tests, including a single-leg stance on both legs with eyes open and closed. Two balance indexes regarding the angular velocities of the waist and chest were defined to assess postural stability. The gait asymmetry indexes of yoga instructors were significantly lower than those of the typically developed controls. Similarly, the yoga instructors had better body balance in all four single-leg stance tests. This study's findings suggest that yoga improves gait asymmetry and balance ability in healthy adults. In the future, further intervention studies could be conducted to confirm the effect of yoga training.

Superior gait performance and balance ability in Latin dancers.

Background: Latin dance consists of various fast and stability-challenging movements that require constant body adjustments to maintain proper posture and balance. Although human gaits are assumed to be symmetrical, several factors can contribute to asymmetrical behavior of the lower extremities in healthy adults. These include lower limb dominance, ground reaction forces, lower limb muscle power, foot placement angle, and range of joint motion. Gait impairment can lead to a high risk of falling, diminished mobility, and even cognition impairment. We hypothesized that Latin dancers might have a more symmetric gait pattern and better balance ability than healthy non-dancer controls. Methods: We investigated the impact of Latin dance training on gait behaviors and body balance. We recruited twenty Latin dancers and 22 normal healthy subjects to conduct walking experiments and one-leg stance tests, and we measured their kinematic data by inertial measurement units. We then defined four performance indexes to assess gait performance and body stability to quantify the potential advantages of dance training. Results: We found that the two gait asymmetric indexes during the walking test and the two performance indexes during the one-leg stance tests were better in Latin dancers compared with the healthy control group. The results confirmed the superiority of Latin dancers over the healthy control group in gait symmetry and balance stability. Our results suggest that Latin dancing training could effectively strengthen lower limb muscles and core muscle groups, thereby improving coordination and enhancing gait performance and balance. Conclusion: Latin dance training can benefit gait performance and body balance. Further studies are needed to investigate the effect of Latin dance training on gait and balance outcomes in healthy subjects and patients with gait disorders.

Interleukins and rheumatoid arthritis: bi-directional Mendelian randomization investigation.

OBJECTIVES: To assess the causality of the associations between interleukins (ILs) and rheumatoid arthritis (RA) using Mendelian randomization (MR) design. METHODS: Genetic instruments and summary-level data for ten ILs were obtained from three genome-wide association meta-analyses. Corresponding data on RA were obtained from a meta-analysis of 22 genome-wide association studies (14,361 cases and 43,923 controls) and the FinnGen consortium (6236 cases, 4596 seropositive cases, 1937 seronegative cases, and 172,834 controls). Forward and reverse MR analyses were performed. RESULTS: The odds ratios (ORs) of RA were 2.08 (95% confidence interval (CI), 1.56-2.77; p<0.001), 2.14 (95% CI, 1.85-2.49; p<0.001), and 0.95 (95% CI, 0.92-0.97; p<0.001) for one standard deviation increase in genetically predicted IL-1ß, IL-6 and IL-6 receptor antagonist (IL-6ra) levels, respectively. There were suggestive associations of genetically predicted IL-1 receptor antagonist (IL-1ra) (OR, 0.85, 95% CI, 0.76, 0.96; p=0.010) and IL-18 (OR, 1.07, 95% CI, 1.00, 1.15; p=0.043) levels with RA risk. Subtype-specific associations were observed for seropositive RA (IL-1ß, IL-1ra, and IL-6) and seronegative RA (IL-2 receptor alpha subunit, IL-8, and IL-18). Reverse MR analysis found a suggestive association between genetic liability to RA and IL-6 receptor antagonist (change 0.015; 95% CI, 0.003-0.028; p=0.015). CONCLUSIONS: This MR study suggests that long-term IL-1 and IL-6 inhibition may reduce the risk of RA, particularly seropositive RA. Upregulations of ILs involved in IL-6 signaling pathways appears to be downstream effects of RA, which supports the blocking IL-6 treatment for RA.

CpG-C ODN M362 as an immunoadjuvant for HBV therapeutic vaccine reverses the systemic tolerance against HBV.

Chronic Hepatitis B virus (CHB) infection is a global public health problem. Oligodeoxynucleotides (ODNs) containing class C unmethylated cytosine-guanine dinucleotide (CpG-C) motifs may provide potential adjuvants for the immunotherapeutic strategy against CHB, since CpG-C ODNs stimulate both B cell and dendritic cell (DC) activation. However, the efficacy of CpG-C ODN as an anti-HBV vaccine adjuvant remains unclear. In this study, we demonstrated that CpG M362 (CpG-C ODN) as an adjuvant in anti-HBV vaccine (cHBV-vaccine) successfully and safely eliminated the virus in HBV-carrier mice. The cHBV-vaccine enhanced DC maturation both in vivo and in vitro, overcame immune tolerance, and recovered exhausted T cells in HBV-carrier mice. Furthermore, the cHBV-vaccine elicited robust hepatic HBV-specific CD8+ and CD4+ T cell responses, with increased cellular proliferation and IFN-γ secretion. Additionally, the cHBV-vaccine invoked a long-lasting follicular CXCR5+ CD8+ T cell response following HBV re-challenge. Taken together, CpG M362 in combination with rHBVvac cleared persistent HBV and achieved long-term virological control, making it a promising candidate for treating CHB.

A core-shell structured COVID-19 mRNA vaccine with favorable biodistribution pattern and promising immunity.

Although inoculation of COVID-19 vaccines has rolled out globally, there is still a critical need for safe and effective vaccines to ensure fair and equitable supply for all countries. Here, we report on the development of a highly efficacious mRNA vaccine, SW0123 that is composed of sequence-modified mRNA encoding the full-length SARS-CoV-2 Spike protein packaged in core-shell structured lipopolyplex (LPP) nanoparticles. SW0123 is easy to produce using a large-scale microfluidics-based apparatus. The unique core-shell structured nanoparticle facilitates vaccine uptake and demonstrates a high colloidal stability, and a desirable biodistribution pattern with low liver targeting effect upon intramuscular administration. Extensive evaluations in mice and nonhuman primates revealed strong immunogenicity of SW0123, represented by induction of Th1-polarized T cell responses and high levels of antibodies that were capable of neutralizing not only the wild-type SARS-CoV-2, but also a panel of variants including D614G and N501Y variants. In addition, SW0123 conferred effective protection in both mice and non-human primates upon SARS-CoV-2 challenge. Taken together, SW0123 is a promising vaccine candidate that holds prospects for further evaluation in humans.

Detection and Classification of Stroke Gaits by Deep Neural Networks Employing Inertial Measurement Units.

This paper develops Deep Neural Network (DNN) models that can recognize stroke gaits. Stroke patients usually suffer from partial disability and develop abnormal gaits that can vary widely and need targeted treatments. Evaluation of gait patterns is crucial for clinical experts to make decisions about the medication and rehabilitation strategies for the stroke patients. However, the evaluation is often subjective, and different clinicians might have different diagnoses of stroke gait patterns. In addition, some patients may present with mixed neurological gaits. Therefore, we apply artificial intelligence techniques to detect stroke gaits and to classify abnormal gait patterns. First, we collect clinical gait data from eight stroke patients and seven healthy subjects. We then apply these data to develop DNN models that can detect stroke gaits. Finally, we classify four common gait abnormalities seen in stroke patients. The developed models achieve an average accuracy of 99.35% in detecting the stroke gaits and an average accuracy of 97.31% in classifying the gait abnormality. Based on the results, the developed DNN models could help therapists or physicians to diagnose different abnormal gaits and to apply suitable rehabilitation strategies for stroke patients.

Functional monocytic myeloid-derived suppressor cells increase in blood but not airways and predict COVID-19 severity.

The immunopathology of coronavirus disease 2019 (COVID-19) remains enigmatic, causing immunodysregulation and T cell lymphopenia. Monocytic myeloid-derived suppressor cells (M-MDSCs) are T cell suppressors that expand in inflammatory conditions, but their role in acute respiratory infections remains unclear. We studied the blood and airways of patients with COVID-19 across disease severities at multiple time points. M-MDSC frequencies were elevated in blood but not in nasopharyngeal or endotracheal aspirates of patients with COVID-19 compared with healthy controls. M-MDSCs isolated from patients with COVID-19 suppressed T cell proliferation and IFN-γ production partly via an arginase 1-dependent (Arg-1-dependent) mechanism. Furthermore, patients showed increased Arg-1 and IL-6 plasma levels. Patients with COVID-19 had fewer T cells and downregulated expression of the CD3ζ chain. Ordinal regression showed that early M-MDSC frequency predicted subsequent disease severity. In conclusion, M-MDSCs expanded in the blood of patients with COVID-19, suppressed T cells, and were strongly associated with disease severity, indicating a role for M-MDSCs in the dysregulated COVID-19 immune response.

Histone demethylase PHF8 drives neuroendocrine prostate cancer progression by epigenetically upregulating FOXA2.

Neuroendocrine prostate cancer (NEPC) is a more aggressive subtype of castration-resistant prostate cancer (CRPC). Although it is well established that PHF8 can enhance prostate cancer cell proliferation, whether PHF8 is involved in prostate cancer initiation and progression is relatively unclear. By comparing the transgenic adenocarcinoma of the mouse prostate (TRAMP) mice with or without Phf8 knockout, we systemically examined the role of PHF8 in prostate cancer development. We found that PHF8 plays a minimum role in initiation and progression of adenocarcinoma. However, PHF8 is essential for NEPC because not only is PHF8 highly expressed in NEPC but also animals without Phf8 failed to develop NEPC. Mechanistically, PHF8 transcriptionally upregulates FOXA2 by demethylating and removing the repressive histone markers on the promoter region of the FOXA2 gene, and the upregulated FOXA2 subsequently regulates the expression of genes involved in NEPC development. Since both PHF8 and FOXA2 are highly expressed in NEPC tissues from patients or patient-derived xenografts, the levels of PHF8 and FOXA2 can either individually or in combination serve as NEPC biomarkers and targeting either PHF8 or FOXA2 could be potential therapeutic strategies for NEPC treatment. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.

Phenotyping of Myeloid-Derived Suppressor Cells in Nonhuman Primates.

While myeloid-derived suppressor cells (MDSCs) in humans and mice have been intensively investigated, there is limited knowledge of these cells in nonhuman primates (NHPs). NHPs serve as critical models for late-stage testing of several biomedical inventions before proceeding with clinical trials and it is therefore important to fully understand their immune compartments and similarities with humans. Here, using antihuman cross-reactive antibodies, we provide flow cytometric analysis protocols for identification of MDSCs in the blood of rhesus macaques, one of the major NHP species as experimental models. Discrepancies and similarities between rhesus and human MDSCs are discussed.