Soybean oil containing ginseng saponins as adjuvants promotes production of cytokines and enhances immune responses to foot-and-mouth disease vaccine.

In the present study, the adjuvant effect of soybean oil containing ginseng root saponins (SO-GS-R) on the immune response to foot-and-mouth disease vaccine (FMDV) in mice was investigated. When immunized with FMDV antigen emulsified in an SO-GS-R formulation, mice generated remarkably higher serum antibody and cytokine responses than mice immunized with FMDV antigen alone. To elucidate the mechanisms underlying the adjuvant effect of SO-GS-R, we measured cytokines in serum and muscle tissue after intramuscular injection of SO-GS-R. The results showed that injection of SO-GS-R significantly increased the levels of IL-1ß, IL-5, IL-6, G-CSF, KC, MCP-1, MIP-1α, and MIP-1ß in both serum and muscle. These results suggested that SO-GS-R recruits neutrophils, eosinophils, T cells and macrophages, causing immune cell recruitment at the injection site, driving antigen-presenting cells to actively participate in the onset of immunity, and amplifying the immune responses. Considering its adjuvant activity and plant-derived properties, SO-GS-R should be further studied for its adjuvant effect on vaccines used in food animals.

A manganese superoxide dismutase (MnSOD) from ark shell, Scapharca broughtonii: Molecular characterization, expression and immune activity analysis.

Manganese superoxide dismutase (MnSOD) is one of the key members of the antioxidant defense enzyme family, however, data regarding to the immune function of MnSOD in mollusks still remain limited now. In this study, a full-length MnSOD cDNA was identified by rapid amplification of cDNA ends (RACE) method from cDNA library of ark shell Scapharca broughtonii (termed SbMnSOD). The cDNA contained an open reading frame (ORF) of 696 bp which encoded a polypeptide of 232 amino acids, a 5'-UTR with length of 32 bp and a 3'-UTR of 275 bp. Four putative amino acid residues (His-57, His-105, Asp-190 and His-194) responsible for manganese coordination were located in the most highly conserved regions of SbMnSOD and the signature sequence (DVWEHAYY) also existed in SbMnSOD. The deduced amino acid sequence of SbMnSOD shared high homology to MnSOD from other species. All those data revealed that the SbMnSOD was a novel member of the MnSOD family. The mRNA expression profiles of SbMnSOD in tissues of foot, gill, mantle, adductor muscle, hemocytes and hepatopancreas analyzed by quantitative real-time PCR (qRT-PCR) suggested the mRNA transcripts of SbMnSOD distributed in all the examined tissues. Importantly, Vibrio anguillarum challenge resulted in the increased expression of SbMnSOD mRNA with a regular change trend in all examined tissues, indicating SbMnSOD actively participated in the immune response process. What's more, further analysis on the antibacterial activity of the recombinant SbMnSOD showed that the fusion protein could remarkably inhibit growth of both Gram-positive and Gram-negative bacteria. The present results clearly suggested that SbMnSOD was an acute phase protein involved in the immune reaction in S. broughtonii.

Omega-3 fatty acids reduce adipose tissue macrophages in human subjects with insulin resistance.

Fish oils (FOs) have anti-inflammatory effects and lower serum triglycerides. This study examined adipose and muscle inflammatory markers after treatment of humans with FOs and measured the effects of ω-3 fatty acids on adipocytes and macrophages in vitro. Insulin-resistant, nondiabetic subjects were treated with Omega-3-Acid Ethyl Esters (4 g/day) or placebo for 12 weeks. Plasma macrophage chemoattractant protein 1 (MCP-1) levels were reduced by FO, but the levels of other cytokines were unchanged. The adipose (but not muscle) of FO-treated subjects demonstrated a decrease in macrophages, a decrease in MCP-1, and an increase in capillaries, and subjects with the most macrophages demonstrated the greatest response to treatment. Adipose and muscle ω-3 fatty acid content increased after treatment; however, there was no change in insulin sensitivity or adiponectin. In vitro, M1-polarized macrophages expressed high levels of MCP-1. The addition of ω-3 fatty acids reduced MCP-1 expression with no effect on TNF-α. In addition, ω-3 fatty acids suppressed the upregulation of adipocyte MCP-1 that occurred when adipocytes were cocultured with macrophages. Thus, FO reduced adipose macrophages, increased capillaries, and reduced MCP-1 expression in insulin-resistant humans and in macrophages and adipocytes in vitro; however, there was no measureable effect on insulin sensitivity.

Effects of dehydration on immune functions after a judo practice session.

We investigated the effects of dehydration after a judo practice session on player muscle and immune functions. Subjects included 25 female university judoists. Investigations were performed before and after 2.5 h of regular judo practice. Body composition, serum enzymes (myogenic enzymes, immunoglobulins and complements), neutrophils counts, reactive oxygen species (ROS) production capability, and phagocytic activity (PA) were measured. Subjects were divided into two groups according to level of dehydration after practice (mild dehydration and severe dehydration groups) and results were compared. Creatine kinase was found to increase significantly after practice. In addition, neutrophil count also increased significantly after practice in both groups. The changing ratios of IgA, IgG and C3 observed in the mild dehydration group were significantly higher than those in the severe dehydration group. In the severe dehydration group, post-practice PA/neutrophil had decreased significantly. Significant positive correlations were found between severity of dehydration and changing ratios of IgA, IgG, IgM, C3, C4 and ROS production capabilities, whereas no significant association was seen with PA and/or serum SOD activity. These results suggest that dehydration resulted in immunosuppression, including decreased neutrophil function.

Adjuvant System AS03 containing α-tocopherol modulates innate immune response and leads to improved adaptive immunity.

AS03 is an Adjuvant System (AS) containing α-tocopherol and squalene in an oil-in-water (o/w) emulsion. AS03 has been considered for the development of pandemic and seasonal influenza vaccines. Key features of AS03's mode of action were investigated in vivo in mice and ex vivo in human cells. AS03's adjuvant activity was superior to that of aluminium hydroxide and required the spatio-temporal co-localisation of AS03 with the antigen. This requirement coincided with AS03 triggering a transient production of cytokines at the injection site and in the draining lymph nodes (dLNs). The nature of the cytokines produced was consistent with the enhanced recruitment of granulocytes and of antigen-loaded monocytes in the dLNs. The presence of α-tocopherol in AS03 was required for AS03 to achieve the highest antibody response. The presence of α-tocopherol also modulated the expression of some cytokines, including CCL2, CCL3, IL-6, CSF3 and CXCL1; increased the antigen loading in monocytes; and increased the recruitment of granulocytes in the dLNs. Hence, AS03's promotion of monocytes as the principal antigen-presenting cells, and its effects on granulocytes and cytokines, may all contribute to enhancing the antigen-specific adaptive immune response.

Ce-TZP/Al2O3 nanocomposite as a bearing material in total joint replacement.

The objectives of this study were to investigate the biocompatibility, phase stability, and wear properties of a newly developed Ce-TZP/Al(2)O(3) nanocomposite, as compared to conventional ceramics, and to determine whether the new composite could be used as a bearing material in total joint prostheses. In tests of mechanical properties, this composite showed significantly higher toughness than conventional Y-TZP. For biocompatibility tests, cylindrical specimens of both the Ce-TZP/Al(2)O(3) nanocomposite and monolithic alumina were implanted into the paraspinal muscles of male Wistar rats. The tissue reactions were almost the same, and at 24 weeks after implantation, thin fibrous capsules with almost no inflammation were observed around both of them. There were no significant differences in membrane thickness between the two ceramics. After hydrothermal treatment in 121 degrees C vapor for 18 h, the new composite showed complete resistance to aging degradation, whereas Y-TZP showed a phase transformation of 25.3 vol% (initial 0.4%) to the monoclinic form. According to the results of pin-on-disk tests, the wear rates of Ce-TZP/Al(2)O(3) nanocomposite and alumina were 0.55 +/- 0.04 x 10(-7) and 2.12 +/- 0.37 x 10(-7)mm(3)/Nm, respectively. The results of this study suggest that the Ce-TZP/Al(2)O(3) nanocomposite is a promising alternative ceramic component for total joint replacement.

Autoantibodies to calpastatin (an endogenous inhibitor for calcium-dependent neutral protease, calpain) in systemic rheumatic diseases.

We identified an autoantibody that reacts with calpastatin [an inhibitor protein of the calcium-dependent neutral protease calpain (EC 3.4.22.17)]. In early immunoblot studies, sera from patients with rheumatoid arthritis (RA) recognized unidentified 60-, 45-, and 75-kDa proteins in HeLa cell extracts. To identify these autoantigens, we used patient sera to clone cDNAs from a lambda gt11 expression library. We isolated clones of four genes that expressed fusion proteins recognized by RA sera. The 1.2-kb cDNA insert (termed RA-6) appeared to encode a polypeptide corresponding to the 60-kDa antigen from HeLa cells, since antibodies bound to the RA-6 fusion protein also reacted with a 60-kDa HeLa protein. The deduced amino acid sequence of the RA-6 cDNA was completely identical with the C-terminal 178 amino acids of human calpastatin except for one amino acid substitution. Patient sera that reacted with the RA-6 also bound pig muscle calpastatin, and a monoclonal antibody to human calpastatin recognized the RA-6 fusion protein, confirming the identity of RA-6 with calpastatin. Moreover, the purified RA-6 fusion protein inhibited the proteolytic activity of calpain, and IgG from a serum containing anti-calpastatin antibodies blocked the calpastatin activity of the RA-6 fusion protein. Immunoblots of the RA-6 product detected autoantibodies to calpastatin in 57% of RA patients; this incidence was significantly higher than that observed in other systemic rheumatic diseases, including systemic lupus erythematosus (27%), polymyositis/dermatomyositis (24%), systemic sclerosis (38%), and overlap syndrome (29%). Thus, anti-calpastatin antibodies are present most frequently in patients with RA and may participate in pathogenic mechanisms of rheumatic diseases.

Complementary surface epitopes, myotropic adhesion and active grip in Trypanosoma cruzi-host cell recognition.

Plasma membranes with complementary surface epitopes and with essentially the same orientation as tissue infective metacyclic trypomastigotes and epimastigotes of Trypanosoma cruzi adhere to L6 myoblast host cells preferentially to smooth muscle and epithelial cells as a function of time, surface area and concentration in saturation phenomena at 4 and 37 degrees C. The initial adhesion rates are partially calcium ion dependent and, at saturation, they are also dependent on high energy phosphorylated intermediates, exerting an active grip on adherent parasite membranes. These phenomena are consistent with the existence of parasite attachment molecules on the external surface of the plasma membrane and complementary host cell receptor structures with the capacity to bind them.

Glutamate-like immunoreactivity in identified neuronal populations of insect nervous systems.

Glutamate is considered to be the most likely transmitter candidate at excitatory synapses onto skeletal muscles of insects. We investigated the distribution of glutamate-like immunoreactivity (Glu-LI) in identified motor neurons of glutaraldehyde-fixed metathoracic ganglia of the locust in paraffin serial sections. The presumably glutamatergic fast and slow extensor tibiae motor neurons show Glu-LI, whereas other cells, including the GABAergic common inhibitory motor neurons and the cluster of octopaminergic dorsal unpaired median cells, show rather low levels of staining. Immunoreactivity of the fast extensor tibiae motor neuron is located in soma, neurites, axon, and the terminal arborizations. A double-labeling experiment on sections of the locust metathoracic ganglion showed that antisera against glutamate and GABA discriminate between the presumably glutamatergic and GABAergic motor neurons and that GABA-LI-positive neurons are low in Glu-LI. The results suggest that Glu-LI can be used as a marker for detecting potential glutamatergic neurons in insects under the present conditions. Application of the glutamate antiserum to sections of the honeybee brain revealed Glu-LI in motor neurons but also in certain interneurons. The most prominent populations of Glu-LI-positive cells were the monopolar cells and large ocellar interneurons, which are first-order interneurons of the visual and ocellar system. Several groups of descending interneurons also showed Glu-LI. The distributions of Glu-LI and GABA-LI are complementary in locust and bee ganglia. The high level of Glu-LI in certain interneuronal populations, as well as in identified glutamatergic motor neurons, suggests that insect central nervous systems may contain glutamatergic neuronal pathways.

A postnatal change in the immunological properties of the acetylcholine receptor at rat muscle endplates.

We have used a myasthenic serum that in adult rat muscle is specific for acetylcholine receptors (AChRs) in the extra-junctional membrane to characterize the AChRs at developing endplates. Immunocytochemical experiments show that AChRs at endplates in the rat diaphragm bind the myasthenic antibodies during the first week after birth but lose their reactivity during the second and third postnatal weeks. AChRs at endplates in adult rat diaphragm do not bind the antibodies even after denervation; in contrast, AChRs at endplates in an adult chicken muscle (anterior latissimus dorsi) are recognized by the antibodies. The loss of immunological reactivity thus may be correlated with a change in the channel properties of the AChR and with the appearance of synaptic folds, two postnatal developmental changes that occur at the endplates of rats, but not of chickens.

Immunogenicity of fatty acid anilides in rabbits and the pathogenesis of the Spanish toxic oil syndrome.

Fatty acid anilides, the major xenobiotic found in the cooking oils responsible for the Spanish toxic oil syndrome, are immunogenic for rabbits as ascertained by a skin test reaction, the characterization of specific antibodies against anilides and the immunofluorescent detection of 'anilide dependent antigens' in tissue slices from treated animals.

Effect of hypothalamic lesions on immediate hypersensitivity.

Bilateral electrolytic lesions were placed in the anterior basal hypothalamus of male Hartley-strain guinea pigs. The animals with anterior hypothalamic lesions and a group of sham-operated controls were sensitized 5-10 days postoperatively with ovalbumin in modified Freund's adjuvant. At 18-22 days following sensitization both groups were randomly divided into three subgroups and challenged by intrajugular injections of three different antigen doses. Anaphylactic death and the severity of anaphylaxis in the survivors were recorded. Antibody titers were determined prior to challenge with use of the passive cutaneous anaphylaxis technique. Serial sections of each brain were made for lesion localization. Significant protection against anaphylaxis was found in guina pigs with symmetrical lesions in the anterior hypothalamus. No significant differences in antibody titers were observed between the lesion and sham-operated groups. There were no significant differences in the response of isolated muscle strips from anterior-lesioned and sham-operated guinea pigs to exogenous histamine. Anterior hypothalamic lesions of size and location comparable to those that provided protection against lethal anaphylaxis did not modify the anaphylactic response of isolated ileum from actively sensitized animals or following passive sensitization in vitro.