Von Willebrand Factor Facilitates Intravascular Dissemination of Microsporidia Encephalitozoon hellem.

Microsporidia are a group of spore-forming, fungus-related pathogens that can infect both invertebrates and vertebrates including humans. The primary infection site is usually digestive tract, but systemic infections occur as well and cause damages to organs such as lung, brain, and liver. The systemic spread of microsporidia may be intravascular, requiring attachment and colonization in the presence of shear stress. Von Willebrand Factor (VWF) is a large multimeric intravascular protein and the key attachment sites for platelets and coagulation factors. Here in this study, we investigated the interactions between VWF and microsporidia Encephalitozoon hellem (E. hellem), and the modulating effects on E. hellem after VWF binding. Microfluidic assays showed that E. hellem binds to ultra-large VWF strings under shear stress. In vitro germination assay and infection assay proved that E. hellem significantly increased the rates of germination and infection, and these effects would be reversed by VWF blocking antibody. Mass spectrometry analysis further revealed that VWF-incubation altered various aspects of E. hellem including metabolic activity, levels of structural molecules, and protein maturation. Our findings demonstrated that VWF can bind microsporidia in circulation, and modulate its pathogenicity, including promoting germination and infection rate. VWF facilitates microsporidia intravascular spreading and systemic infection.

Characterization of a Murine Model for Encephalitozoon hellem Infection after Dexamethasone Immunosuppression.

BACKGROUND: Encephalitozoon hellem (E. hellem) belongs to a group of opportunistic pathogens called microsporidia. Microsporidia infection symptoms vary and include diarrhea, ocular disorders and systemic inflammations. Traditionally, immunodeficient animals were used to study microsporidia infection. To overcome the difficulties in maintenance and operation using immunodeficient mice, and to better mimic natural occurring microsporidia infection, this study aims to develop a pharmacologically immunosuppressed murine model of E. hellem infection. METHODS: Wild-type C57BL/6 mice were immunosuppressed with dexamethasone (Dex) and then E. hellem spores were inoculated into the mice intraperitoneally. Control groups were the Dex-immunosuppressed but noninoculated mice, and the Dex-immunosuppressed then lipopolysaccharide (LPS)-treated mice. Mice body weights were monitored and all animals were sacrificed at the 15th day after inoculation. Tissue fragments and immune cells were collected and processed. RESULTS: Histopathological analysis demonstrated that E. hellem inoculation resulted in a disseminated nonlethal infection. Interestingly, E. hellem infection desensitized the innate immunity of the host, as shown by cytokine expressions and dendritic cell maturation. We also found that E. hellem infection greatly altered the composition of host gut microbiota. (4) Conclusions: Dex-immunosuppressed mice provide a useful tool for study microsporidiosis and the interactions between microsporidia and host immunity.

Hemocytin facilitates host immune responses against Nosema bombycis.

Invertebrates lack an adaptive immune response and thus are reliant on their innate immune response for eliminating invading pathogens. The innate immune responses of silkworms against the pathogen Nosema bombycis include: hemocyte aggregation, melanization, antimicrobial peptides, etc. In our current study, we discovered that a silkworm hemostasis-related protein, hemocytin, is up-regulated after Nosema bombycis infection. This novel finding lead to our hypothesis that hemocytin participates in immune responses against N. bombycis. We investigated this hypothesis by analyzing the adhesive effects of hemocytin to invading N. bombycis, and the hemocytin-mediated hemocyte aggregation and hemolymph melanization. We showed that hemocytin can adhere to the surface of N. bombycis, which facilitates the agglutination of N. bombycis and hemocytes as well as the subsequent melanization. Moreover, when we utilize RNAi technology to decrease in vivo hemocytin expression, we found that the proliferation of N. bombycis within the host significantly increased. These results support our hypothesis that hemocytin exerts pro-inflammatory effects by facilitating pathogen agglutination, along with hemocyte aggregation and melanization, to combat N. bombycis. Our study is the first to determine a function of hemocytin in innate immunity against N. bombycis. Moreover, our findings are of great importance to provide potential targets for developing novel strategy against microsporidia infection.