A multi-epitope subunit vaccine based on CU/ZN-SOD, OMP31 and BP26 against Brucella melitensis infection in BALB/C mice.

Brucellosis, a zoonosis caused by Brucella, is highly detrimental to both humans and animals. Most existing vaccines are live attenuated vaccines with safety flaws for people and animals. Therefore, it is advantageous to design a multi-epitope subunit vaccine (MEV) to prevent Brucella infection. To this end, we applied a reverse vaccinology approach. Six cytotoxic T cell (CTL) epitopes, seven T helper cell (HTL) epitopes, and four linear B cell epitopes from CU/ZN-SOD, Omp31, and BP26 were obtained. We linked the CTL, HTL, B-cell epitopes, the appropriate CTB molecular adjuvant, and the universal T helper lymphocyte epitope, PADRE, with linkers AAY, GPPGG, and KK, respectively. This yielded a 412-amino acid MEV construct, which we named MEVcob. The immunogenicity, stability, safety, and feasibility of the construct were evaluated by bioinformatics tools (including the AlphaFold2 prediction tool, the AlphaFold2 tool, NetMHC-I pan 4.0 server, IEDB MHC-I server, ABCpred service, and C-ImmSim server); the physicochemical properties, secondary and tertiary structures, and binding ability of MEVocb to toll-like receptor 4 (TLR4) was analyzed. Then, codon adaptation and computer cloning studies were performed. MEVocb is highly immunogenic in immunostimulation experiments, The proteins translated by these sequences were relatively stable, exhibiting a high antigenic index. Furthermore, mouse experiments confirmed that the MEVocb construct could raise IFN-γ, IgG, IgG2a, IgG1, IL-2, TNF-α levels in mice, indicating that induced a specific humoral and cellular immune response in BALB/c mice. This vaccine induced a statistically significant level of protection in BALB/c mice when challenged with Brucella melitensis 043 in Xinjiang. Briefly, we utilized immunoinformatic tools to design a novel multi-epitope subunit candidate vaccine against Brucella. This vaccine aims to induce host immune responses and confer specific protective effects. The study results offer a theoretical foundation for the development of a novel Brucella subunit vaccine.

[Protective effect of haliotidis on the oxidative damage in the human lens epithelial cells].

OBJECTIVE: To study the influence of haliotidis extractive on the oxidative damage in the human lens epithelial cells cultured in vitro. METHODS: Experimental study. Cultured human lens epithelial cells in vitro were intervened with hydrogen peroxide caused oxidative damage model, at the same time added different concentrations of concha haliotidis extractive. With control experiment research cells were divided into the blank control group, positive control group hydrogen peroxide group and hydrogen peroxide and different concentrations of concha haliotidis group, and on the first, third, fifth day the activity of Cultured human lens epithelial cells were detected with Cell Counting Kit-8 (CCK-8) , cellular proliferation and morphological changes were observed with interred phase contrast microscope, and then on the third day chemical colorimetric were used to detect the homogenates superoxide dismutase(SOD), glutathione(GSH) and malondialdehyde (MDA) level. RESULTS: (1) At different time points there were variations between the activity of HLEC in each experimental group, Among each experimental group HLEC OD value of the cell vitality at 1 d, 3 d, 5 d , respectively were blank control group: 0.88, 1.28, 1.32; Positive control group: 0.73, 1.02, 1.06; 0.001% concha haliotis extract group: 0.73, 1.03, 1.06; 0.01% concha haliotis extract group: 0.76, 1.10, 1.13; 0.1% concha haliotis extract group: 0.79, 1.22, 1.21; 0.3% concha haliotis extract group: 0.79, 1.21, 1.21; the difference between groups was statistically significant (P < 0.05) (1 d, F = 23 922.42, P < 0.05;3 d, F = 120 605.86, P < 0.05; 5 d, F = 150 939.45, P < 0.05). H2O2 made the vitality of the cells reduce, concha haliotidis enhance its vitality, and in a certain range of time and concentrations there was dependence, with which the third day and 0.1% was the best. (2) After adding H2O2, the SOD and GSH level of HLEC reduced,(SOD 158.05 U/mgprot,GSH 15.05 mg/gprot) but MDA increased to 18.11 nmol/mgprot, concha haliotidis groups made the increase of antioxidant level(SOD 188.64 U/mgprot,GSH 21.05 mg/1000 mgprot)and the decrease of lipid peroxidation in oxidative damaged HLECs(MDA 14.16 nmol/mgprot), change had a statistical significance(P < 0.05) (SOD: F = 983.04, P < 0.05; GSH: F = 444.44, P < 0.05; MDA: F = 830.52, P < 0.05). (3)The chromatin of the positive control group concentrated and aggregated obviously, the aggregation of chromatin in concha haliotidis group lightened. CONCLUSION: The concha haliotidis can protect the cultured human lens epithelial cells in vitro which are oxidative injured, increased intracellular antioxidant levels, reduce the generation of hazardous products.

Characterization and complete genome sequence of human coronavirus NL63 isolated in China.

Human coronavirus NL63 (HCoV-NL63) was first discovered in Amsterdam in 2004 and was identified as a new human respiratory coronavirus. We here report the first complete genome sequence of HCoV-NL63 strain CBJ 037 isolated in 2008 from a patient with bronchitis in Beijing, China.

Human Coronaviruses HCoV-NL63 and HCoV-HKU1 in Hospitalized Children with Acute Respiratory Infections in Beijing, China.

The human coronaviruses (HCoVs) HCoV-NL63 and HCoV-HKU1 are two recently discovered coronaviruses that circulate widely and are associated with acute respiratory infections (ARI). We detected HCoV-NL63 and HCoV-HKU1 in specimens collected from May 2008 to March 2010 from patients with ARI aged <7.75 years of age attending the Beijing Children's Hospital. Thirty-two (8.4%) and 57 (14.9%) of 382 specimens tested positive for HCoV-NL63 and HCoV-HKU1, respectively, by real-time RT-PCR. Use of a Luminex xTAG RVP Fast kit showed that coinfection with respiratory syncytial virus and parainfluenza 3 virus was common among patients infected with either virus type. In HCoV-HKU1-infected patients, the predominant clinical symptoms were cough, fever, and expectoration. In HCoV-NL63-infected patients they were cough, fever, and rhinorrhea. Phylogenetic studies showed that the HCoV-HKU1 nucleoprotein gene was relatively conserved compared to NCBI reference sequences, while the 1ab gene of HCoV-NL63 showed more variation.