Construction of recombinant Lactococcus expressing thymosin and interferon fusion protein and its application as an immune adjuvant.

BACKGROUND: In recent years, biosafety and green food safety standards have increased the demand for immune enhancers and adjuvants. In the present study, recombinant food-grade Lactococcus lactis (r-L. lactis-Tα1-IFN) expressing thymosin Tα1 and chicken interferon fusion protein was constructed. RESULTS: The in vitro interactions with macrophages revealed a mixture of recombinant r-L. lactis-Tα1-IFN could significantly activate both macrophage J774-Dual™ NF-κB and interferon regulator (IRF) signaling pathways. In vitro interactions with chicken peripheral blood mononuclear cells (PBMCs) demonstrated that a mixture of recombinant r-L. lactis-Tα1-IFN significantly enhanced the expression levels of interferon (IFN)-γ, interleukin (IL)-10, CD80, and CD86 proteins in chicken PBMCs. Animal experiments displayed that injecting a lysis mixture of recombinant r-L. lactis-Tα1-IFN could significantly activate the proliferation of T cells and antigen-presenting cells in chicken PBMCs. Moreover, 16S analysis of intestinal microbiota demonstrated that injection of the lysis mixture of recombinant r-L. lactis-Tα1-IFN could significantly improve the structure and composition of chicken intestinal microbiota, with a significant increase in probiotic genera, such as Lactobacillus spp. Results of animal experiments using the lysis mixture of recombinant r-L. lactis-Tα1-IFN as an immune adjuvant for inactivated chicken Newcastle disease vaccine showed that the serum antibody titers of the experimental group were significantly higher than those of the vaccine control group, and the expression levels of cytokines IFN-γ and IL-2 were significantly higher than those of the vaccine control group. CONCLUSION: These results indicate that food-safe recombinant r-L. lactis-Tα1-IFN has potential as a vaccine immune booster and immune adjuvant. This study lays the foundation for the development of natural green novel animal immune booster or immune adjuvant.

The Phase Equilibria of Natural Gas Hydrate in the Presence of 1,3-Dimethylcyclohexane and Octyl-ß-D-glucopyranoside.

The thermodynamic effect of octyl-ß-D-glucopyranoside (OGP) on the formation of methane-1,3-dimethylcyclohexane (DMCH) hydrate was studied in this work. The thermodynamic equilibrium hydrate formation pressures between 275.15 K and 283.15 K were measured by the isothermal pressure search method. Different OGP aqueous solutions (0, 0.1, and 1 wt%) were used in this work. The experimental results show that OGP had no obvious thermodynamic inhibition on methane-DMCH hydrate formation when its concentration was low (0.1 wt%), whereas it had an inhibition on methane-DMCH hydrate formation when its concentration was high (1 wt%). The phase equilibrium hydrate formation pressure of the methane-DMCH-OGP system is about 0.1 MPa higher than that of the methane-DMCH system. The dissociation enthalpies of methane hydrate in different solutions remained uniform, which indicates that OGP was not involved in methane-DMCH hydrate formation. This phenomenon is explained from the perspective of the molecular structure of OGP. As a renewable and biological nonionic surfactant, the concentration of OGP in the liquid phase is low, so OGP can be added to the methane-DMCH system without significant thermodynamic inhibition.

Experimental and Modeling Study on Methane Hydrate Equilibrium Conditions in the Presence of Inorganic Salts.

The aim of this study was to determine the influence of four inorganic salts, KCl, NaCl, KBr and NaBr, on the thermodynamic conditions of methane hydrate formation. In order to achieve this, the vapor-liquid water-hydrate (VLWH) equilibrium conditions of methane (CH4) hydrate were measured in the temperature range of 274.15 K-282.15 K by the isothermal pressure search method. The results demonstrated that, in comparison with deionized water, the four inorganic salts exhibited a significant thermodynamic inhibition on CH4 hydrate. Furthermore, the inhibitory effect of Na+ on methane hydrate is more pronounced than that of K+, where there is no discernible difference between Cl- and Br-. The dissociation enthalpy (∆Hdiss) of CH4 hydrate in the four inorganic salt solutions is comparable to that of deionized water, indicating that the inorganic salt does not participate in the formation of hydrate crystals. The Chen-Guo hydrate model and N-NRTL-NRF activity model were employed to forecast the equilibrium conditions of CH4 hydrate in electrolyte solution. The absolute relative deviation (AARD) between the predicted and experimental values were 1.24%, 1.08%, 1.18% and 1.21%, respectively. The model demonstrated satisfactory universality and accuracy. This study presents a novel approach to elucidating the mechanism and model prediction of inorganic salt inhibition of hydrate.

Enterococcus faecium C171: Modulating the Immune Response to Acute Lethal Viral Challenge.

Commensal bacteria modulate acute immune responses to infection in hosts. In this study, Enterococcus faecium C171 was screened and isolated. This strain has similar basic characteristics to the reference probiotic, including strong anti-inflammatory and anti-infective effects. E. faecium C171 inhibits the production of pro-Caspase-1 and significantly reduces the production of interleukin-1ß (IL-1ß) in vitro. These reactions were confirmed using the Transwell system. Live E. faecium C171 mainly exerted an inhibitory effect on acute inflammation, whereas the anti-infective and immune-activating effects were primarily mediated by the E. faecium C171-produced bacterial extracellular vesicles (Efm-C171-BEVs). Furthermore, in the specific pathogen-free (SPF) chicken model, oral administration of E. faecium C171 increased the relative abundance of beneficial microbiota (Enterococcus and Lactobacillus), particularly Enterococcus, the most important functional bacteria of the gut microbiota. E. faecium C171 significantly inhibited the acute inflammatory response induced by a highly virulent infectious disease, and reduced mortality in SPF chickens by 75%. In addition, E. faecium C171 induced high levels of CD3+, CD4-, and CD8- immunoregulatory cells and CD8+ killer T cells, and significantly improved the proliferative activity of T cells in peripheral blood mononuclear cells, and the secretion of interferon-γ. These findings indicate that E. faecium C171 and Efm-C171-BEVs are promising candidates for adjuvant treatment of acute inflammatory diseases and acute viral infections.