Tuft cells:a new flavor in innate epithelial immunity

How host cells sense intestinal parasitic infection and initiate the appropriate immune response has long been a focus of many immunologists. Three new papers now identify a critical role for tuft cells, an epithelial cell type involved in perception of taste, as key players that kick-start type 2 immunity

Immunity to Trichinella Spiralis

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Adaptive immunity alters distinct host feeding pathways during nematode induced inflammation, a novel mechanism in parasite expulsion

Gastrointestinal infection is often associated with hypophagia and weight loss; however, the precise mechanisms governing these responses remain poorly defined. Furthermore, the possibility that alterations in feeding during infection may be beneficial to the host requires further study. We used the nematode Trichinella spiralis, which transiently inhabits the small intestine before migrating to skeletal muscle, as a biphasic model of infection to determine the cellular and molecular pathways controlling feeding during enteric and peripheral inflammation. Through the infection of genetically modified mice lacking cholecystokinin, Tumor necrosis factor α receptors and T and B-cells, we observed a biphasic hypophagic response to infection resulting from two separate immune-driven mechanisms. The enteroendocrine I-cell derived hormone cholecystokinin is an essential mediator of initial hypophagia and is induced by CD4+ T-cells during enteritis. In contrast, the second hypophagic response is extra-intestinal and due to the anorectic effects of TNFα during peripheral infection of the muscle. Moreover, via maintaining naive levels of the adipose secreted hormone leptin throughout infection we demonstrate a novel feedback loop in the immunoendocrine axis. Immune driven I-cell hyperplasia and resultant weight loss leads to a reduction in the inflammatory adipokine leptin, which in turn heightens protective immunity during infection. These results characterize specific immune mediated mechanisms which reduce feeding during intestinal or peripheral inflammation. Importantly, the molecular mediators of each phase are entirely separate. The data also introduce the first evidence that I-cell hyperplasia is an adaptively driven immune response that directly impinges on the outcome to infection

The mucus barrier : immune defence against gastrointestinal nematodes

Trichuriasis, caused by the intestinal nematode Trichuris, is a disease that affects up to a billion people worldwide. To date, most of our knowledge of this disease comes from the mouse model, Trichuris muris, which has been successfully used to dissect the immune-mediated effector mechanisms that elicit the expulsion of the nematode. Numerous studies have shown a temporal association between intestinal nematode expulsion and goblet cell hyperplasia; however their precise role in response to nematode infection remains elusive. Goblet cells found at mucosal surfaces secrete many constituent components of the mucus barrier, including the gel-forming mucins (Muc2 in the intestine); mucins are large multifunctional glycoproteins that provide the structural framework of the barrier. The studies presented in this thesis demonstrate that the mucosal barrier and in particular its mucin components, changes in response to acute and chronic T.muris infection. In animals resistant to chronic T. muris infection, IL-13-mediated increase in Muc2 production and secretion was observed at the site of infection. Critically, expulsion of the nematode was significantly delayed in the absence of Muc2. Further investigation subsequently showed that Muc5ac, a mucin normally expressed in non-intestinal mucosa was, in fact, expressed in the intestine following nematode infection and was associated with nematode expulsion in the resistant mice. Moreover, mice deficient in Muc5ac were susceptible to chronic infection, despite a strong underlying TH2-type immune response which is essential to eliminate the nematodes, suggesting that Muc5ac acts as a critical effector molecule. Several qualitative changes in the mucins were also noted during resistance: mucins were more highly charged and more sulphated during nematode expulsion. Overall, the changes within the mucus barrier during resistance result in altering the rheological properties of the mucus layer making it less porous and mucins were shown to directly 'damage' the nematodes during nematode expulsion as reflected by a significant reduction in ATP levels. Chronic infection was accompanied by decreased levels of low charged and highly sialylated Muc2. Additionally, we demonstrated that the excretory secretory products released by the nematode consist of serine proteases capable of depolymerising the Muc2 mucin network, which may be part of the nematodes regime to improve its niche and/or aid movement through the mucus layer. Overall, this resulted in a porous mucus layer and a favourable environment for the parasite.Data is presented to show that the intestinal mucus barrier and its constituent mucins are an integral part of the co-ordinated expulsion mechanisms that occur in animals resistant to T.muris infection and we identify a mechanism whereby the nematode exerts its effects on the mucin environment to promote its survival within the host.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Modulation of dendritic cells and autoimmunity by apoptotic and necrotic cells

As the principal antigen-presenting cells to T cells, dendritic cells (DCs) have a key role in the balance of immunity and autoimmunity. They are essential in two major, converse roles - eliciting T cell immune responses to pathogenic material, and maintaining peripheral tolerance to self-tissue by inhibiting self-reactive T cells. These functions involve the processing of pathogenic or self antigens and subsequent presentation of antigenic peptides on MHC to antigen-specific T cells. DC recognition of conserved pathogenic markers induces a mature phenotype that governs immunogenic presentation to T cells and, consequently, the adaptive immune response. In contrast, DC recognition of self tissue suppresses maturation, instead inducing a tolerogenic phenotype that induces self antigen-specific T cell to die, become anergised, or converted to T regulatory cells. Apoptotic cells are the major source of self-antigen for the maintenance of peripheral tolerance, and their defective clearance by DCs is implicated in autoimmunity. Apoptotic cells are thought to actively suppress maturation of DCs and inhibit the possible immune responses promoted by proinflammatory mediators released from necrotic cells. However, the immune function of apoptotic cells and their relative influence over necrotic cells are highly contested, partially due to the complex nature of immunogenicity arising from the sourcing and generation of apoptotic cells. In this investigation, various methods of inducing apoptosis and necrosis are evaluated. Definitive methods of inducing well-characterised cell death are then employed to compare the effects of apoptotic and necrotic cells on dendritic cells and in vitro and in vivo immune responses. Reported here are in vitro findings that support previous reports of the anti-inflammatory response of DCs to apoptotic cells, and the inflammatory response of DCs to necrotic cells. The previously-reported inhibitory effect of apoptotic cells on LPS-induced secretion of Th1 cytokines is supported here, but the inhibitory effect of apoptotic cells on LPS-induced upregulation of co-stimulatory molecules is contested. Novel findings describe the upregulation of DC expression of co-inhibitory molecules induced by both apoptotic cells and necrotic cells. Apoptotic cells, but not necrotic cells, had a suppressive effect on CpG-induced upregulation of co-stimulatory molecules and pro-inflammatory cytokines. Apoptotic cells suppressed the capacity of untreated and CpG-treated, but not LPS-treated, DCs to elicit IFNγ production by T cells. Apoptotic cells, but not necrotic cells, induced regulatory T cells and partially restored their CpG-suppressed induction. Finally, apoptotic cell-modulation of DCs inhibited the induction of autoimmunity in a novel modification of an in vivo model of diabetes. Interestingly, novel evidence for the possibility of necrotic cell-induced tolerance by means of direct T cell killing is addressed.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Whipworm secretions and their roles in host-parasite interactions

Whipworm (Trichuris) is a genus of roundworms that causes gastrointestinal infections in humans and animals. Of particular interest are T. trichiura, the causative agent of human trichuriasis, a neglected tropical disease that affects 477 million people worldwide, and T. suis, the pig whipworm species, responsible for growth stunting and economic losses within the agricultural industry. The naturally occurring mouse whipworm, T. muris, has been used for decades as a model for trichuriasis, yielding knowledge on the biology of these parasites and the host response to infection. Ex vivo culture of T. muris (and to some extent, T. suis) has provided insight into the composition of the excretory/secretory (E/S) products released by worms, which include a myriad of proteins, RNAs, lipids, glycans, metabolites and extracellular vesicles. T. muris E/S has formed the basis of the search for whipworm vaccine candidates, while the immunomodulatory potential of T. suis and T. muris secretions has been investigated with the aim of improving our understanding of how these parasites modulate host immunity, as well as identifying immunomodulatory candidates with therapeutic potential in the context of inflammatory diseases. This article will review the various components found within Trichuris E/S, their potential as vaccine candidates and their immunomodulatory properties. Graphical Abstract: [Figure not available: see fulltext.]

Chronic Trichuris muris infection in C57BL/6 mice causes significant changes in most microbiota and metabolome:effects reversed by pathogen clearance

Trichuris species are a globally important and prevalent group of intestinal helminth parasites, in which Trichuris muris (mouse whipworm) is an ideal model for this disease. This paper describes the first ever highly controlled and comprehensive investigation into the effects of T. muris infection on the faecal microbiota of mice and the effects on the microbiota following successful clearance of the infection. Communities were profiled using DGGE, 454 pyrosequencing, and metabolomics. Changes in microbial composition occurred between 14 and 28 days post infection, resulting in significant changes in α and β- diversity. This impact was dominated by a reduction in the diversity and abundance of Bacteroidetes, specifically Prevotella and Parabacteroides. Metabolomic analysis of stool samples of infected mice at day 41 showed significant differences to uninfected controls with a significant increase in the levels of a number of essential amino acids and a reduction in breakdown of dietary plant derived carbohydrates. The significant reduction in weight gain by infected mice probably reflects these metabolic changes and the incomplete digestion of dietary polysaccharides. Following clearance of infection the intestinal microbiota underwent additional changes gradually transitioning by day 91 towards a microbiota of an uninfected animal. These data indicate that the changes in microbiota as a consequence of infection were transitory requiring the presence of the pathogen for maintenance. Interestingly this was not observed for all of the key immune cell populations associated with chronic T. muris infection. This reflects the highly regulated chronic response and potential lasting immunological consequences of dysbiosis in the microbiota. Thus infection of T. muris causes a significant and substantial impact on intestinal microbiota and digestive function of mice with affects in long term immune regulation

IL-17A both initiates, via IFNγ suppression, and limits the pulmonary type 2 immune response to nematode infection

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