Isoprinosine as a foot-and-mouth disease vaccine adjuvant elicits robust host defense against viral infection through immunomodulation.

Background: Commercial foot-and-mouth disease (FMD) vaccines have limitations, such as local side effects, periodic vaccinations, and weak host defenses. To overcome these limitations, we developed a novel FMD vaccine by combining an inactivated FMD viral antigen with the small molecule isoprinosine, which served as an adjuvant (immunomodulator). Method: We evaluated the innate and adaptive immune responses elicited by the novel FMD vaccine involved both in vitro and in vivo using mice and pigs. Results: We demonstrated isoprinosine-mediated early, mid-term, and long-term immunity through in vitro and in vivo studies and complete host defense against FMD virus (FMDV) infection through challenge experiments in mice and pigs. We also elucidated that isoprinosine induces innate and adaptive (cellular and humoral) immunity via promoting the expression of immunoregulatory gene such as pattern recognition receptors [PRRs; retinoic acid-inducible gene (RIG)-I and toll like receptor (TLR)9], transcription factors [T-box transcription factor (TBX)21, eomesodermin (EOMES), and nuclear factor kappa B (NF-kB)], cytokines [interleukin (IL)-12p40, IL-23p19, IL-23R, and IL-17A)], and immune cell core receptors [cluster of differentiation (CD)80, CD86, CD28, CD19, CD21, and CD81] in pigs. Conclusion: These findings present an attractive strategy for constructing novel FMD vaccines and other difficult-to-control livestock virus vaccine formulations based on isoprinosine induced immunomodulatory functions.

Bestatin, A Pluripotent Immunomodulatory Small Molecule, Drives Robust and Long-Lasting Immune Responses as an Adjuvant in Viral Vaccines.

An inactivated whole-virus vaccine is currently used to prevent foot-and-mouth disease (FMD). Although this vaccine is effective, it offers short-term immunity that requires regular booster immunizations and has several side effects, including local reactions at the vaccination site. To address these limitations, herein, we evaluated the efficacy of bestatin as a novel small molecule adjuvant for inactivated FMD vaccines. Our findings showed that the FMD vaccine formulated with bestatin enhanced early, intermediate-, and particularly long-term immunity in experimental animals (mice) and target animals (pigs). Furthermore, cytokines (interferon (IFN)α, IFNß, IFNγ, and interleukin (IL)-29), retinoic acid-inducible gene (RIG)-I, and T-cell and B-cell core receptors (cluster of differentiation (CD)28, CD19, CD21, and CD81) markedly increased in the group that received the FMD vaccine adjuvanted with bestatin in pigs compared with the control. These results indicate the significant potential of bestatin to improve the efficacy of inactivated FMD vaccines in terms of immunomodulatory function for the simultaneous induction of potent cellular and humoral immune response and a long-lasting memory response.

The role of zinc sulfate in enhancing cellular and humoral immune responses to foot-and-mouth disease vaccine.

Foot-and-mouth disease (FMD) is a rapidly propagating infectious disease of cloven-hoofed animals, especially cattle and pigs, affecting the productivity and profitability of the livestock industry. Presently, FMD is controlled and prevented using vaccines; however, conventional FMD vaccines have several disadvantages, including short vaccine efficacy, low antibody titers, and safety issues in pigs, indicating the need for further studies. Here, we evaluated the efficacy of a novel bivalent vaccine containing zinc sulfate as an immunostimulant and FMD type O and A antigens (O PA2 and A YC, respectively) against FMD virus in mice and pigs. Zinc sulfate induced cellular immunity in murine peritoneal exudate cells (PECs) and porcine peripheral blood mononuclear cells (PBMCs) by increasing IFNγ secretion. Additionally, FMD vaccine containing O PA2 and A YC antigens and zinc sulfate induced early, mid-, and long-term immune responses in mice and pigs, and enhanced cellular and humoral immunity by regulating the expression of pathogen recognition receptors (PRRs), transcription factors, co-stimulatory molecules, and cytokines in porcine PBMCs from vaccinated pigs. Overall, these results indicated that the novel immunostimulant zinc sulfate induced potent cellular and humoral immune responses by stimulating antigen-presenting cells (APCs) and T and B cells, and enhanced long-term immunity by promoting the expression of co-stimulatory molecules. These outcomes suggest that zinc sulfate could be used as a novel vaccine immunostimulant for difficult-to-control viral diseases, such as African swine fever (ASF) or COVID-19.

Caesalpinia sappan L. ameliorates hypercholesterolemia in C57BL/6 mice and suppresses inflammatory responses in human umbilical vein endothelial cells (HUVECs) by antioxidant mechanism.

Oxidative stress and inflammatory mediators were measured in the plasma and livers of C57BL/6 mice fed a high-cholesterol diet for 14 weeks and in cultured human umbilical vein endothelial cells (HUVECs). Some of the mice fed with the atherogenic diet received drinking water supplemented with 0.01 g of a 70% ethanol extract of Caesalpinia sappan L. (CSLE) per 20 g of body weight. Numerous parameters were determined: concentrations of total, high-, and low-density cholesterol; atherogenic index; plasma trolox equivalent antioxidant capacity (TEAC); levels of hepatic thiobarbituric acid reactive substances (TBARS) and protein carbonyls; and the activities of hepatic antioxidant enzymes, including Cu·Zn-SOD, Mn-SOD, glutathione peroxidase, glutathione reductase, and catalase. HUVECs were stimulated with tumor necrosis factor α (TNFα) and the expression of intracellular reactive oxygen species (ROS), lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), adhesion molecules, inhibitory κBα (IκBα), and nuclear factor κB (NFκB) were measured. Compared to mice fed a hypercholesterolemic diet alone, mice fed a hypercholesterolemic diet supplemented with CSLE exhibited decreased total plasma cholesterol and increased high-density lipoprotein cholesterol, and thus a lower atherogenic index. Furthermore, plasma TEAC and levels of hepatic TBARS and protein carbonyls were significantly decreased in CSLE-supplemented mice (P < 0.05), whereas all hepatic antioxidative indicators were significantly elevated (P < 0.05). In HUVECs stimulated with TNFα, CSLE significantly decreased the expression of intracellular ROS, LOX-1, and adhesion molecules; the degradation of IκBα; and the nuclear translocation of NFκB; in contrast, CSLE induced the expression of Nrf2 and HO-1 (P < 0.05 for all results).

Leonurus sibiricus herb extract suppresses oxidative stress and ameliorates hypercholesterolemia in C57BL/6 mice and TNF-alpha induced expression of adhesion molecules and lectin-like oxidized LDL receptor-1 in human umbilical vein endothelial cells.

In Leonurus sibiricus herb extract (LHE)-supplemented animals, plasma cholesterol decreased and high-density lipoprotein-cholesterol increased, resulting in a lowered atherogenic index. The plasma trolox equivalent antioxidant capacity, levels of hepatic thiobarbituric acid-reactive substances, and protein carbonyl values decreased significantly in LHE-supplemented mice (p<0.05), whereas the hepatic antioxidant indicators were all significantly elevated (p<0.05). In human umbilical vein endothelial cells stimulated with tumor necrosis factor alpha, LHE significantly suppressed intracellular reactive oxygen species, LOX-1, and adhesion molecules. LHE supplementation may modulate the lipoprotein composition and attenuate oxidative stress by elevated antioxidant processes, thus suppressing the activation of inflammatory mediators. This is a possible mechanism of the anti-atherogenic effect.

Curcumin inhibits TNFalpha-induced lectin-like oxidised LDL receptor-1 (LOX-1) expression and suppresses the inflammatory response in human umbilical vein endothelial cells (HUVECs) by an antioxidant mechanism.

In this study, the anti-oxidative activities of 70% ethanol extract from Curcuma aromatica Salisb. (CAS) and curcumin (CUR) were studied. The CAS extracts and CUR were both found to have a potent scavenging activity against the reactive species tested, as well as an inhibitory effect on LDL oxidation. Cultured human umbilical vein endothelial cells (HUVECs) were stimulated with tumour necrosis factor alpha (TNFalpha), expression of intracellular reactive oxygen species (ROS), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), lectin-like oxidised LDL receptor-1 (LOX-1), adhesion molecules, inhibitory kappa Balpha (IkappaBalpha) and nuclear factor kappa B (NFkappaB) were measured. In HUVECs stimulated with TNFalpha, CUR significantly suppressed expression of the intracellular ROS, LOX-1 and adhesion molecules, degradation of IkappaBalpha and translocation of NFkappaB, while inducing production of NO by phosphorylation of eNOS (p <0.05). In conclusion, CAS and CUR may modulate lipoprotein composition and attenuate oxidative stress by elevated antioxidant processes.

Antioxidant properties of benzylchroman derivatives from Caesalpinia sappan L. against oxidative stress evaluated in vitro.

The antioxidant activity of extracts from Caesalpinia sappan L. (CSL) was studied in vitro by evaluating the total phenolics, measuring the antioxidant activity by TEAC, measuring the scavenging effects on reactive oxygen species (ROS) and on reactive nitrogen species (RNS), and measuring the inhibitory effect on Cu(2+)-induced human low-density lipoprotein (LDL) oxidation. The CSL extracts were found to have a potent scavenging activity against all of the reactive species tested, as well as an inhibitory effect on LDL oxidation, especially in the ethyl acetate (EA) fraction. Therefore, we isolated and identified benzylchroman derivatives sappanchalcone (1) and 3'-deoxy-4-O-methylepisappanol (2) from the EA fraction of CSL and their antioxidant activities were evaluated. The studied CSL extracts and the compounds 1 and 2 were revealed to be very effective against the evaluated pro-oxidant species, including ROS and RNS.

Protective effects of luteolin-7-O-beta-D-glucuronide methyl ester from the ethyl acetate fraction of Lycopi Herba against pro-oxidant reactive species and low-density lipoprotein peroxidation.

In this study the potent scavenging activity of "Lycopi Herba" (LH) extract was studied using the following: evaluation of the total phenolics, measuring the antioxidant activity by Trolox equivalent antioxidant concentration, measuring the scavenging effects on reactive oxygen species, on reactive nitrogen species, and measuring the inhibitory effect on Cu(2+) induced human low-density lipoprotein oxidation in vitro. The ethyl acetate fraction from the LH extracts were found to have a potent scavenging activity against all of the reactive species tested, as well as an inhibitory effect on LDL oxidation. Therefore, we isolated and identified luteolin-7-O-beta-D-glucuronide methyl ester as the major compound from the ethyl acetate fraction of LH and their antioxidant activities were evaluated.

Effect of bamboo culm extract on oxidative stress and genetic expression: bamboo culm extract ameliorates cell adhesion molecule expression and NFkappaB activity through the suppression of the oxidative stress.

BACKGROUND & AIMS: This study was designed to investigate whether bamboo culm extract (BCE) supplementation may ameliorate risk factors of cardiovascular diseases, such as hypercholesterolemia. METHODS: Oxidative stress and inflammatory mediators in plasma, livers of C57BL/6 mice fed high-cholesterol diet and calf pulmonary artery endothelial (CPAE) cells. Briefly, C57BL/6 mice were fed the high-cholesterol diet which was supplemented with 1% (w/w), or 3% (w/w) of BCE for 16 weeks. The concentration of total cholesterol, LDL-cholesterol, HDL-cholesterol level and atherogenic index were measured. Plasma TEAC value, hepatic thiobarbituric acid reactive substances (TBARS), protein carbonyl values and hepatic antioxidant enzyme activities, such as Cu,Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase (GSH-Px), GSH reductase and catalase were determined. In addition, hepatic nuclear factor kappa B activities were detected. In the calf pulmonary artery endothelial (CPAE) cells stimulated with lipopolysaccharide, the expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1) were measured. RESULTS: Plasma cholesterol level was decreased, while HDL-cholesterol was increased, thus atherogenic index was lowered in BCE-supplemented animals. Plasma trolox equivalent and hepatic thiobarbituric acid reactive substances and protein carbonyl values were lowered significantly in BCE groups (p<0.05) in a dose-dependent manner. Hepatic antioxidative enzyme activities, such as Cu,Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase (GSH-P), GSH reductase, and catalase were elevated in mice fed BCE-supplemented diets (p<0.05). Nuclear factor kappa B activities of livers and vascular cell adhesion molecule-1 and intracellular cell adhesion molecule-1 expressions in CPAE cells stimulated with lipopolysaccharide were significantly lowered in BCE groups (p<0.05). CONCLUSION: These results suggest that BCE supplementation may modulate lipoprotein composition and attenuate oxidative stress by elevated antioxidative processes, thus suppressing inflammatory mediator activation as possible mechanism of its anti-atherogenic effect.

Bamboo culm extract supplementation elevates HDL-cholesterol and ameliorates oxidative stress in C57BL/6 mice fed atherogenic diet.

Previously, we reported that bamboo culms possess a stronger antioxidative capacity than bamboo leaves in vitro. In this study, we investigated whether bamboo culm extract (BCE) supplementation ameliorates oxidative stress and hepatic nuclear factor kappaB (NF kappa B) activation in C57BL/6 mice fed an atherogenic diet. In addition, the effect of BCE supplementation on plasma lipid levels of the animals was tested. The mice were randomly assigned to a normal diet, an atherogenic diet (control), or an atherogenic diet supplemented with 1% (wt/wt) BCE or 3% (wt/wt) BCE for 16 weeks. Atherogenic diet-induced oxidative stress, measured by hepatic thiobarbituric acid-reactive substances and protein carbonyls, was significantly lower in the BCE-supplemented groups than in the control (P < .05). Total antioxidative capacity was elevated in the BCE groups, along with greater activities of antioxidative enzymes such as superoxide dismutase and catalase, compared to the control or normal groups (P < .05). The hepatic NF kappa B binding activities were significantly lower in the BCE groups as well (P < .05). The high-density lipoprotein-cholesterol level was significantly elevated by BCE supplementation (P < .05), whereas the effects of BCE on triglyceride and total cholesterol were inconsistent. Results from this study suggest that BCE supplementation may lessen oxidative stress via a series of changes, including a reinforced antioxidant system, and also suggest that the lowered oxidative stress status may down-regulate the activation of inflammatory mediators.