Alteration of the Thymic T Cell Repertoire by Rotavirus Infection Is Associated with Delayed Type 1 Diabetes Development in Non-Obese Diabetic Mice

Rotaviruses are implicated as a viral trigger for the acceleration of type 1 diabetes in children. Infection of adult non-obese diabetic (NOD) mice with rotavirus strain RRV accelerates diabetes development, whereas RRV infection in infant NOD mice delays diabetes onset. In this study of infant mice, RRV titers and lymphocyte populations in the intestine, mesenteric lymph nodes (MLN) and thymus of NOD mice were compared with those in diabetes-resistant BALB/c and C57BL/6 mice. Enhanced intestinal RRV infection occurred in NOD mice compared with the other mouse strains. This was associated with increases in the frequency of CD8αβ TCRαβ intraepithelial lymphocytes, and their PD-L1 expression. Virus spread to the MLN and T cell numbers there also were greatest in NOD mice. Thymic RRV infection is shown here in all mouse strains, often in combination with alterations in T cell ontogeny. Infection lowered thymocyte numbers in infant NOD and C57BL/6 mice, whereas thymocyte production was unaltered overall in infant BALB/c mice. In the NOD mouse thymus, effector CD4⁺ T cell numbers were reduced by infection, whereas regulatory T cell numbers were maintained. It is proposed that maintenance of thymic regulatory T cell numbers may contribute to the increased suppression of inflammatory T cells in response to a strong stimulus observed in pancreatic lymph nodes of adult mice infected as infants. These findings show that rotavirus replication is enhanced in diabetes-prone mice, and provide evidence that thymic T cell alterations may contribute to the delayed diabetes onset following RRV infection.     

Identification of Annexin A1 interacting proteins in chronic myeloid leukemia KCL22 cells

In the present study, we used a functional proteomic approach to identify Annexin A1 (Anxa1) interacting proteins in the Philadelphia‐positive KCL22 cell line. We focused on Anxa1 because it is one of the major proteins upregulated in imatinib‐sensitive KCL22S cells versus imatinib‐resistant KCL22R. Our proteomic strategy revealed 21 interactors. Bioinformatic analysis showed that most of these proteins are involved in cell death processes. Among the proteins identified, we studied the interaction of Anxa1 with two phosphatases, Shp1 and Shp2, which were recently identified as biomarkers of imatinib sensitivity in patients affected by chronic myeloid leukemia. Our data open new perspectives in the search for annexin‐mediated signaling pathways and may shed light on mechanisms of resistance to imatinib that are unrelated to Bcr‐Abl activity. All mass spectrometry data have been deposited in the ProteomeXchange with identifier PXD000030.     

Rotavirus Activates Lymphocytes from Non-Obese Diabetic Mice by Triggering Toll-Like Receptor 7 Signaling and Interferon Production in Plasmacytoid Dendritic Cells

It has been proposed that rotavirus infection promotes the progression of genetically-predisposed children to type 1 diabetes, a chronic autoimmune disease marked by infiltration of activated lymphocytes into pancreatic islets. Non-obese diabetic (NOD) mice provide a model for the human disease. Infection of adult NOD mice with rhesus monkey rotavirus (RRV) accelerates diabetes onset, without evidence of pancreatic infection. Rather, RRV spreads to the pancreatic and mesenteric lymph nodes where its association with antigen-presenting cells, including dendritic cells, induces cellular maturation. RRV infection increases levels of the class I major histocompatibility complex on B cells and proinflammatory cytokine expression by T cells at these sites. In autoimmunity-resistant mice and human mononuclear cells from blood, rotavirus-exposed plasmacytoid dendritic cells contribute to bystander polyclonal B cell activation through type I interferon expression. Here we tested the hypothesis that rotavirus induces bystander activation of lymphocytes from NOD mice by provoking dendritic cell activation and proinflammatory cytokine secretion. NOD mouse splenocytes were stimulated with rotavirus and assessed for activation by flow cytometry. This stimulation activated antigen-presenting cells and B cells independently of virus strain and replicative ability. Instead, activation depended on virus dose and was prevented by blockade of virus decapsidation, inhibition of endosomal acidification and interference with signaling through Toll-like receptor 7 and the type I interferon receptor. Plasmacytoid dendritic cells were more efficiently activated than conventional dendritic cells by RRV, and contributed to the activation of B and T cells, including islet-autoreactive CD8⁺ T cells. Thus, a double-stranded RNA virus can induce Toll-like receptor 7 signaling, resulting in lymphocyte activation. Our findings suggest that bystander activation mediated by type I interferon contributes to the lymphocyte activation observed following RRV infection of NOD mice, and may play a role in diabetes acceleration by rotavirus.     

Novel strains of Bacillus,isolated from compost and compost-amended soils,as biological control agents against soil-borne phytopathogenic fungi

A stepwise screening strategy made it possible to identify five new Bacillus spp. strains for biocontrol of Rhizoctonia solani, Sclerotinia minor and Fusarium solani. In vitro and in vivo biocontrol activity and M13-PCR DNA-fingerprinting led to the selection of these valuable biological control agents (BCAs) from a wide collection of over 250 candidates. At the end of this selection, the highest potential antagonists were identified at species level by 16S-rRNA gene sequence analysis, and results assigned them to Bacillus subtilis group as Bacillus amyloliquefaciens- and Bacillus methylotrophicus-related strains. In the current study, spore-forming bacteria provided substantial biocontrol of telluric diseases on cress and other different host plants. The strains named 15S and 09C were effective in disease control on Brassica oleracea/R. solani pathosystem, whereas Sclerotinia drop of lettuce was reduced by treatments with the strains 17S and 08C. Finally, the strains 17S and 12S were equally effective to control potato Fusarium rot. The evident zone of inhibition seen in dual culture plates suggested antibiosis-like antagonisms as the main mechanisms used by these bacterial isolates in interaction with the pathogens. Additionally, the API-ZYM method revealed constitutive activity of certain extracellular enzymes that could be involved in plant fortification. Bacillus strains isolated from compost and compost-amended soils are promising BCAs that have potential for practical application as biofungicides.     

Humic substances influence sodium metabolism in the freshwater crustacean Daphnia magna

Humic substances are ubiquitous components of natural waters with important roles in alleviating metal toxicity to aquatic organisms. Recent literature reports suggest that humic substances may also exert direct influences on biota. This study investigated the influence of two commercially available humic substances on sodium metabolism in Daphnia magna, a hyperregulating freshwater crustacean. Environmentally realistic levels of Suwannee River natural organic matter (SRN) and Aldrich humic acid (AHA) significantly enhanced sodium transport. This effect was described as an uncompetitive stimulation of sodium influx, as characterised by an increased maximal sodium transport rate (Jmax), accompanied by a decreased uptake affinity (increased Km). SRN exposure also significantly promoted the unidirectional loss of sodium from the daphnids to the water, an effect not observed in the presence of AHA. A 24-h preexposure to AHA before influx measurement had no effect on AHA-induced stimulation of sodium influx. Conversely, 24-h preexposure to SRN resulted in influx values that returned to control (humic-free) levels. Whole-body sodium levels reduced by SRN exposure were also restored to control levels following 24-h SRN preexposure. The significance and potential mechanisms of these actions are discussed, and the toxicological implications of these findings are assessed.     

Evolutionary conservation of Ceratitis capitata transformer gene function

Transformer functions as a binary switch gene in the sex determination and sexual differentiation of Drosophila melanogaster and Ceratitis capitata, two insect species that separated nearly 100 million years ago. The TRA protein is required for female differentiation of XX individuals, while XY individuals express smaller, presumably nonfunctional TRA peptides and consequently develop into adult males. In both species, tra confers female sexual identity through a well-conserved double-sex gene. However, unlike Drosophila tra, which is regulated by the upstream Sex-lethal gene, Ceratitis tra itself is likely to control a feedback loop that ensures the maintenance of the female sexual state. The putative CcTRA protein shares a very low degree of sequence identity with the TRA proteins from Drosophila species. However, in this study we show that a female-specific Ceratitis Cctra cDNA encoding the putative full-length CcTRA protein is able to support the female somatic and germline sexual differentiation of D. melanogaster XX; tra mutant adults. Although highly divergent, CcTRA can functionally substitute for DmTRA and induce the female-specific expression of both Dmdsx and Dmfru genes. These data demonstrate the unusual plasticity of the TRA protein that retains a conserved function despite the high evolutionary rate. We suggest that transformer plays an important role in providing a molecular basis for the variety of sex-determining systems seen among insects.