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Abstract: The development of hepatic fibrosis is the principal cause of morbidity and mortality in human beings infected with schistosoma. In this study, we investigated the effect of polyinosinic–polycytidylic acid (poly I:C) on Schistosoma japonicum (S. japonicum) egg-induced liver fibrosis. S. japonicum cercariae infected mice were injected with poly I:C at the onset of egg granuloma formation (early phase poly I:C treatment) or after the formation of liver fibrosis (late phase poly I:C treatment). Our results showed that both early and late phase poly I:C treatment significantly reduced collagen deposition and hepatic stellate cell activation in the liver. Poly I:C is one of the most effective adjuvants for Th1 type responses, and its protective effect on liver fibrosis was accompanied by increased IFN-α, IFN-β, IFN-γ, IL-12, TNF-α, and IL-10 mRNA expression, and decreased IL-4 and IL-5 mRNA expression. Moreover, poly I:C injection also enhanced the mRNA expression of natural killer group 2 member D (NKG2D) and tumor necrosis factor related apoptosis-inducing ligand (TRAIL). Therefore, it is indicated that poly I:C can significantly attenuate S. japonicum egg-induced hepatic fibrosis, which may be partly dependent on the increased Th1 response and decreased Th2 response. [Copyright &y& Elsevier]

The role of natural killer (NK) cells in infection-induced liver fibrosis remains obscure. In this study, we elucidated the effect of NK cells on Schistosoma japonicum (S. japonicum) egg-induced liver fibrosis. Liver fibrosis was induced by infecting C57BL/6 mice with 18–20 cercariae of S. japonicum. Anti-ASGM1 antibody was used to deplete NK cells. Toll-like receptor 3 ligand, polyinosinic-polycytidylic acid (poly I∶C) was used to enhance the activation of NK cells. Results showed that NK cells were accumulated and activated after S. japonicum infection, as evidenced by the elevation of CD69 expression and IFN-γ production. Depletion of NK cells markedly enhanced S. japonicum egg-induced liver fibrosis. Administration of poly I∶C further activated NK cells to produce IFN-γ and attenuated S. japonicum egg-induced liver fibrosis. The observed protective effect of poly I∶C on liver fibrosis was diminished through depletion of NK cells. Disruption of IFN-γ gene enhanced liver fibrosis and partially abolished the suppression of liver fibrosis by poly I∶C. Moreover, expression of retinoic acid early inducible 1 (RAE 1), the NKG2D ligand, was detectable at high levels on activated hepatic stellate cells derived from S. japonicum-infected mice, which made them more susceptible to hepatic NK cell killing. In conclusion, our findings suggest that the activated NK cells in the liver after S. japonicum infection negatively regulate egg-induced liver fibrosis via producing IFN-γ, and killing activated stellate cells. Author Summary: Schistosomiasis continues to be a major public health problem in the developing world. Parasite egg-induced liver fibrosis is the principal cause of morbidity and mortality in human infected with schistosoma. Thus, elucidating the mechanisms that restrict tissue fibrosis may lead to more effective strategies for immunological intervention in this and a variety of chronic diseases. NK cells have been demonstrated to play an important role in suppressing carbon tetrachloride (CCl4)-induced liver fibrosis. However, little is known about the role of NK cells in an infection-based model of fibrosis. In the current study, we determined, for the first time, the role of NK cells in S. japonicum egg-induced liver fibrosis. Our findings suggest that the activated NK cells in the liver after S. japonicum infection negatively regulate egg-induced liver fibrosis via producing IFN-γ, and killing activated stellate cells. These results further our understanding of the innate immune cells that regulate the development of S. japonicum-induced fibrosis and aid in the development of potential strategies to enhance immunity against this and other chronic inflammatory diseases of the liver where fibrosis is a common feature. [ABSTRACT FROM AUTHOR]

We report a novel method to analyze multiple cytokines/chemokines in 5 μL of BALF in mice. The methodology relies on micro-volume wall-less immunoassays combined with Luminex magnetic bead multiplex technology. This new method has been demonstrated with good validity, veracity, and sensitivity by comparing it with the existing conventional Luminex method, as well comparing the values of BALF cytokines/chemokines from functional inflammatory stimulated mice with that of healthy mice. Compared with conventional Luminex method, it also showed obvious advantages in some other aspects such as the sample volume was minimized to 5 μL, the consumption of reagents and beads was reduced by a factor of 5 and the total cost of experiments was significantly reduced by 80%, which would have wide applications in the field of multiple cytokines/chemokines analysis. [ABSTRACT FROM AUTHOR]

Background: Entamoeba histolytica is an intestinal protozoan parasite that causes amoebiasis, including amebic dysentery and liver abscesses. E. histolytica invades host tissues by adhering onto cells and phagocytosing them depending on the adaptation and expression of pathogenic factors, including Gal/GalNAc lectin. We have previously reported that E. histolytica possesses multiple CXXC sequence motifs, with the intermediate subunit of Gal/GalNAc lectin (i.e., Igl) as a key factor affecting the amoeba's pathogenicity. The present work showed the effect of immunization with recombinant Igl on amebic liver abscess formation and the corresponding immunological properties. Methodology/Principal Findings: A prokaryotic expression system was used to prepare the full-length Igl and the N-terminal, middle, and C-terminal fragments (C-Igl) of Igl. Vaccine efficacy was assessed by challenging hamsters with an intrahepatic injection of E. histolytica trophozoites. Hamsters intramuscularly immunized with full-length Igl and C-Igl were found to be 92% and 96% immune to liver abscess formation, respectively. Immune-response evaluation revealed that C-Igl can generate significant humoral immune responses, with high levels of antibodies in sera from immunized hamsters inhibiting 80% of trophozoites adherence to mammalian cells and inducing 80% more complement-mediated lysis of trophozoites compared with the control. C-Igl was further assessed for its cellular response by cytokine-gene qPCR analysis. The productions of IL-4 (8.4-fold) and IL-10 (2-fold) in the spleen cells of immunized hamsters were enhanced after in vitro stimulation. IL-4 expression was also supported by increased programmed cell death 1 ligand 1 gene. Conclusions/Significance: Immunobiochemical characterization strongly suggests the potential of recombinant Igl, especially the C-terminal fragment, as a vaccine candidate against amoebiasis. Moreover, protection through Th2-cell participation enabled effective humoral immunity against amebic liver abscesses. [ABSTRACT FROM AUTHOR]

Rapid and high-throughput analytical techniques for IgE that requires a small serum amount are very important, especially for pediatric patients. In these patients, blood is collected from veins, which is painful compared to fingertip blood collection. Herein, a novel microfluidic system capable of high-throughput parallel analyses of allergen-specific IgE from small amounts of patient serum was successfully developed. A six-plex immunoassay was constructed within a microfluidic chip, and the entire system was validated using samples from clinical patients. Major antigens from house dust mite ( Dermatophagoides farinae and Blomia tropicalis ), cat ( Felis domesticus ), fungus ( Cladosporium herbarum ), ragweed ( Humulus japonicas ), and tree pollen ( Platanus acerifolia ) were used as analysis targets. Sample consumption decreased to <0.05 µL compared with the 480 µL serum consumption by fluoroenzyme immunoassay (UniCAP system Pharmacia Diagnostics AB, Uppsala, Sweden), the 50 µL serum consumption by enzyme-linked immune sorbent assay (ELISA), or the 1.5 µL serum consumption by conventional protein chip analysis. Analysis duration, reagent cost, and total cost for each measurement were also considerably decreased. The assay showed good accuracy and sensitivity toward the clinical samples. A significant correlation of allergen-specific IgE levels was found among the microfluidic assay, UniCAP system, and ELISA. [ABSTRACT FROM AUTHOR]