Candida albicans Colonizes and Disseminates to the Gastrointestinal Tract in the Presence of the Microbiota in a Severe Combined Immunodeficient Mouse Model.

Candida albicans is the leading cause of candidemia or other invasive candidiasis. Gastrointestinal colonization has been considered as the primary source of candidemia. However, few established mouse models that mimic this infection route are available. In the present study, we established a mouse model of disseminated candidiasis developed through the translocation of Candida from the gut. In this study, we developed a novel C. albicans GI colonization and dissemination animal model by using severe combined immunodeficient Rag2-/-IL2γc-/- (Rag2γc) mice, which lack functional T, B, NK cells, and IL2γc-dependent signaling. Rag2γc mice were highly susceptible to C. albicans gastrointestinal infection even in the presence of the gut microbiota. Within 4 weeks post infection, Rag2γc mice showed dose-dependent weight loss and disseminated candidiasis in more than 58% (7/12) of moribund mice. Histological analysis demonstrated abundant hyphae penetrating the mucosa, with significant neutrophilic infiltration in mice infected with wild-type C. albicans but not a filamentation-defective mutant. In moribund Rag2γc mice, the necrotic lesions and disrupted epithelial cells were associated with C. albicans hyphae. Notably, removal of the gut microbiota by antibiotics exacerbated the severity of fungal infection in Rag2γc mice, as demonstrated by elevated fungal burdens and accelerated weight loss and death. Furthermore, higher fungal burden and IL-1ß expression were prominently noted in the stomach of Rag2γc mice. In fact, a significant increase in circulating proinflammatory cytokines, including IL-6, TNF-α, and IL-10, indicative of a septic response, was evident in infected Rag2γc mice. Additionally, Rag2γc mice exhibited significantly lower levels of IL-22 but not IFN-γ or IL-17A than wild-type B6 mice, suggesting that IL-22 plays a role in C. albicans gastrointestinal infection. Collectively, our analysis of the Rag2γc mouse model revealed features of C. albicans gastrointestinal colonization and dissemination without the interference from antibiotics or chemotherapeutic agents, thus offering a new investigative tool for delineating the pathogenesis of C. albicans and its cross-talk with the gut microbiota.

Selective activation of inflammation factors by human parvovirus B19 and human bocavirus VP1 unique region on H9c2 cardiomyocyte.

Human parvovirus B19 (B19) and human bocavirus 1 (HBoV) are the only known pathogenic parvoviruses, and are responsible for a variety of diseases in human beings. Mounting evidence indicates a strong association between B19 infection and cardiac disorders including myocarditis, dilated cardiomyopathy and heart failure. However, very limited information about the role of HBoV in cardiac disorders is known. To elucidate the effects of B19 and HBoV on cardiac disorders, we expressed EGFP­conjugate constructs of B19­VP1 unique region (VP1u) and HBoV­VP1u, along with the mutants EGFP­B19­VP1uD175A and EGFP­HBoV­VP1uV12A, in H9c2 cells by stable transfection. The protein expression levels of EGFP, EGFP­B19­VP1u, EGFP­B19­VP1uD175A, EGFP­HBoV­VP1u and EGFP­HBoV­VP1uV12A in H9c2 cells were observed under a fluorescence microscope and confirmed by western blotting. Secreted phospholipase A2 (sPLA2) activity was detected in B19­VP1u and HBoV­VP1u but not B19­VP1uD175A and HBoV­VP1uV12A recombinant proteins. Significantly higher expression levels of MCP2 and IP­10 mRNA were detected in H9c2 cells that were transfected with pEGFP­B19­VP1u, compared with in those cells transfected with pEGFP­HBoV­VP1u, pEGFP­B19­VP1uD175A or pEGFP­HBoV­VP1uV12A. Significantly higher protein levels of IL­1ß and IL­6 were detected in H9c2 cells transfected with pEGFP­B19­VP1u or pEGFP­HBoV­VP1u, compared with in those cells transfected with pEGFP­B19­VP1uD175A or pEGFP­HBoV­VP1uV12A. Notably, significantly higher expression of both TNF­α and NF­κB was observed only in H9c2 cells transfected with pEGFP­B19­VP1u, but not in those cells transfected with pEGFP­HBoV­VP1u, pEGFP­B19­VP1uD175A or pEGFP­HBoV­VP1uV12A. These findings, to our knowledge for the first time, reveal the difference between B19­VP1u and HBoV­VP1u in H9c2 cells and provide insight into the roles of B19­VP1u and HBoV­VP1u in the pathogenesis of cardiac inflammation.