Bacillus subtilis spores as adjuvants against avian influenza H9N2 induce antigen-specific antibody and T cell responses in White Leghorn chickens

Low-pathogenicity avian influenza H9N2 remains an endemic disease worldwide despite continuous vaccination, indicating the need for an improved vaccine strategy. Bacillus subtilis (B. subtilis), a gram-positive and endospore-forming bacterium, is a non-pathogenic species that has been used in probiotic formulations for both animals and humans. The objective of the present study was to elucidate the effect of B. subtilis spores as adjuvants in chickens administered inactivated avian influenza virus H9N2. Herein, the adjuvanticity of B. subtilis spores in chickens was demonstrated by enhancement of H9N2 virus-specific IgG responses. B. subtilis spores enhanced the proportion of B cells and the innate cell population in splenocytes from chickens administered both inactivated H9N2 and B. subtilis spores (Spore + H9N2). Furthermore, the H9N2 and spore administration induced significantly increased expression of the pro-inflammatory cytokines IL-1β and IL-6 compared to that in the H9N2 only group. Additionally, total splenocytes from chickens immunized with inactivated H9N2 in the presence or absence of B. subtilis spores were re-stimulated with inactivated H9N2. The subsequent results showed that the extent of antigen-specific CD4+ and CD8+ T cell proliferation was higher in the Spore + H9N2 group than in the group administered only H9N2. Taken together, these data demonstrate that B. subtilis spores, as adjuvants, enhance not only H9N2 virus-specific IgG but also CD4+ and CD8+ T cell responses, with an increase in pro-inflammatory cytokine production. This approach to vaccination with inactivated H9N2 together with a B. subtilis spore adjuvant in chickens produces a significant effect on antigen-specific antibody and T cell responses against avian influenza virus.This study and medical writing support were funded by Sanofi Genzyme and Regeneron Pharmaceuticals, Inc

Comparing the Effects of Exoskeletal-Type Robot-Assisted Gait Training on Patients with Ataxic or Hemiplegic Stroke

This study aimed to discover the effects of robotic rehabilitation utilizing an exoskeletal-type robot-assisted gait training (RAGT) device on patients with ataxic and hemiplegic stroke and to compare its effectiveness between the two groups. This was a retrospective study, and the electronic charts of 22 patients who underwent RAGT treatment from October 2019 to June 2021 were reviewed. Patients were divided into ataxic and hemiplegic groups based on their symptoms. The clinical outcome measures included the Berg balance scale (BBS), functional ambulation category (FAC), and mobility subcategories of the modified Barthel Index (MBI-m). Outcome measures were reviewed at two points within 48 h, before and after RAGT with EXOWALK®, a type of exoskeletal robot. After the RAGT sessions, total patients in both ataxic and hemiplegic groups demonstrated statistically significant improvements in BBS (p < 0.0001, p = 0.002, and p = 0.005, respectively) and MBI-m (p < 0.0001, p = 0.002, and p = 0.011, respectively). Additionally, FAC after RAGT was significantly improved (p = 0.0056). The regression coefficient of the number of RAGT treatments for BBS changes in the nine subjects was estimated to be 2.45; 3.50 in the ataxic group and 2.26 in the hemiplegic group. The regression coefficient of the number of RAGT treatments for MBI-m changes in the nine subjects was estimated to be 0.16; 4.00 in the ataxic group and −0.52 in the hemiplegic group. Our results suggest that RAGT using an exoskeletal-type robot, EXOWALK®, could be effective for improving walking capacity, balance, and daily activities of life in patients with ataxic and hemiplegic stroke

Microcrystalline Cellulose for Delivery of Recombinant Protein-Based Antigen against Erysipelas in Mice

The study describes the development of a vaccine using microcrystalline cellulose (Avicel PH-101) as a delivery carrier of recombinant protein-based antigen against erysipelas. Recombinant SpaA, surface protective protein, from a gram-positive pathogen Erysipelothrix rhusiopathiae was fused to a cellulose-binding domain (CBD) from Trichoderma harzianum endoglucanase II through a S3N10 peptide. The fusion protein (CBD-SpaA) was expressed in Escherichia coli and was subsequently bound to Avicel PH-101. The antigenicity of CBD-SpaA bound to the Avicel was evaluated by enzyme-linked immunosorbent (ELISA) and confocal laser scanning microscope (CLSM) assays. For the examination of its immunogenicity, groups of mice were immunized with different constructs (soluble CBD-SpaA, Avicel coated with CBD-SpaA, whole bacterin of E. rhusiopathiae (positive control), and PBS (negative control)). In two weeks after immunization, mice were challenged with 1x107 CFU of E. rhusiopathiae and Avicel coated with CBD-SpaA induced protective immunity in mice. In conclusion, this study demonstrates the feasibility of microcrystalline cellulose as the delivery system of recombinant protein subunit vaccine against E. rhusiopathiae infection in mice