ABSTRACT: Alternatively activated mononuclear phagocytes from the skin site of infection and the impact of IL-4Rα signaling on CD4+T cell survival in draining lymph nodes after repeated exposure to Schistosoma mansoni cercariae
Project description:In areas endemic for schistosomiasis, repeated exposure to infective cercariae is a frequent occurrence, and repeated exposure of murine skin to Schistosoma mansoni resulted in CD4+ T cells becoming hypo-responsive. Here potential contributory mechanisms were investigated. In the skin infection site, three mononuclear phagocyte populations were identified (tissue macrophages, dendritic cells, and macrophages) which exhibited up-regulation of genes associated with alternative activation, in particular the gene encoding RELMα. However, in repeatedly infected mice deficient in RELMα, there was no change in the abundance of mononuclear phagocytes in the skin, and CD4+ cells in the skin draining lymph nodes remained hypo-responsive. In mice deficient for IL-4Rα, required for alternative activation, levels of dermal regulatory IL-10 were reduced and there was an increase in the abundance of antigen presenting MHC-IIhigh cells, which was accompanied by increased numbers of CD4+ T cells. Although the absence of IL-4Rα did not translate into increased CD4+ cell responsiveness, they exhibited lower expression of Fas/FasL, resulting in decreased apoptosis/cell death and increased cell viability. This study highlights a mechanism through which IL-4Rα may regulate the immune system through the induction of IL-10 and regulation of Fas/FasL mediated cell death. Mice were infected 1x or 4x with S. mansoni cercariae via the pinnae. Pinnae were harvested 4 days after the final infection and the DEC obtained after an overnight culture in vitro. Cells were then stained with F4/80 APC and MHC-II PE and four different populations sorted (using the MoFlo or the Astrios) based on their expression profiles. Several mice (12-18) from each group were pooled to generate each replicate.
Project description:In areas endemic for schistosomiasis, repeated exposure to infective cercariae is a frequent occurrence, and repeated exposure of murine skin to Schistosoma mansoni resulted in CD4+ T cells becoming hypo-responsive. Here potential contributory mechanisms were investigated. In the skin infection site, three mononuclear phagocyte populations were identified (tissue macrophages, dendritic cells, and macrophages) which exhibited up-regulation of genes associated with alternative activation, in particular the gene encoding RELMα. However, in repeatedly infected mice deficient in RELMα, there was no change in the abundance of mononuclear phagocytes in the skin, and CD4+ cells in the skin draining lymph nodes remained hypo-responsive. In mice deficient for IL-4Rα, required for alternative activation, levels of dermal regulatory IL-10 were reduced and there was an increase in the abundance of antigen presenting MHC-IIhigh cells, which was accompanied by increased numbers of CD4+ T cells. Although the absence of IL-4Rα did not translate into increased CD4+ cell responsiveness, they exhibited lower expression of Fas/FasL, resulting in decreased apoptosis/cell death and increased cell viability. This study highlights a mechanism through which IL-4Rα may regulate the immune system through the induction of IL-10 and regulation of Fas/FasL mediated cell death.
Project description:The mesenteric lymph nodes represent the immune response to eggs in schistosome infections,and the analysis of gene expression profiles of the mesenteric lymph nodes from the Vac-Cha (vaccinated with UV attenuated cercariae and challenged with normal cercariae)and Inf-Con (infected with normal cercariae) groups. We used microarrays to detail the global programme of gene expression and identified distinct classes of up-regulated and down-regulated genes. The mesenteric lymph nodes surrounding the cecum from the Vac-Cha and Inf-Con groups were collected at week 8 post challenge and used to analyze the gene profile with Affymetrix microarrays. We sought to obtain the different genes expression profiles between the Vac-Cha group and Inf-Con group.The Vac-Cha group is three pigs vaccinated with UV attenuated Schistosoma japonicum cercariae and challenged with normal Schistosoma japonicum cercariae, and the Inf-Con group is three pigs infected with normal cercariae. Samples pooled per group. A list of differentially-regulated genes can be found in the supplementary file below.
Project description:We performed V(D)J scRNA sequencing to find transcriptional and B cell receptor repertoire differences in the germinal center of the draining lymph node of mice born to mothers chronically infected with Schistosoma mansoni. Cite-seq was used to help identify antigen specific cells.
Project description:Use of genomic DNA as an indirect reference for identifying gender associated transcripts in morphologically identical, but chromosomally distinct, Schistosoma mansoni cercariae
Project description:The mesenteric lymph nodes represent the immune response to eggs in schistosome infections,and the analysis of gene expression profiles of the mesenteric lymph nodes from the Vac-Cha (vaccinated with UV attenuated cercariae and challenged with normal cercariae)and Inf-Con (infected with normal cercariae) groups. We used microarrays to detail the global programme of gene expression and identified distinct classes of up-regulated and down-regulated genes.
Project description:B cell receptor (BCR)-sequencing from 3 donors with prostate cancer. The sources are Peripheral blood B cells (P), tumour draining lymph nodes (S) and non-draining lymph nodes (N).
Project description:During its life cycle, the helminth parasite Schistosoma mansoni uses the freshwater snail Biomphalaria glabrata as an intermediate host to reproduce asexually generating cercariae for infection of the human definitive host. Following invasion of the snail, the parasite develops from a miracidium to a mother sporocyst and releases excretory secretory products (ESPs) that likely influence the outcome of host infection. To better understand molecular interactions between these ESPs and the host snail defence system, we determined gene expression profiles of haemocytes from S. mansoni resistant or -susceptible strains of B. glabrata exposed in vitro to S. mansoni ESPs (20ug/ml) for 1 h, using a 5K B. glabrata cDNA microarray.
Project description:To determine the influence of primary tumors on pre-metastatic lymph nodes, we have employed whole genome microarray expression profiling as a discovery platform to identify gene signatures of B cells from tumor-draining lymph nodes, compared with normal lymph nodes. We subcutaneously inoculated C57BL/6 mice with the 4T1 mammary carcinoma. Two weeks later, tumor-draining lymph nodes were dissociated and B cells (CD19+) were sorted. Lymph nodes B cells from normal mice without tumor bearing were set as controls.
Project description:To determine the influence of primary tumors on pre-metastatic lymph nodes, we have employed whole genome microarray expression profiling as a discovery platform to identify gene signatures of stromal cells from tumor-draining lymph nodes, compared with normal lymph nodes. We subcutaneously inoculated C57BL/6 mice with the 4T1 mammary carcinoma. Two weeks later, tumor-draining lymph nodes were dissociated and stromal cells (CD45-) were sorted. Lymph nodes stromal cells from normal mice without tumor bearing were set as controls.
