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304 999 at delivery is also associated with elevated levels of endotoxin in maternal and placental plasma. These data suggest additional mechanisms by which AN EX VIVO MODEL FOR STUDYING THE EARLIEST PHASE schistosomiasis negatively impacts the maternal-fetal dyad. OF HEPATIC SCHISTOSOMIASIS 1001 Geoffrey Gobert Queensland Institute of Medical Research, Brisbane, Australia HELMINTH-INDUCED IL-4 ABOLISHES INKT CELL-MEDIATED To investigate the earliest hepatic events associated with the deposition CLEARANCE OF BACTERIURIA of schistosome eggs, we have established a novel technique, involving Yi-Ju Hsieh, Chi-Ling Fu, Michael Hsieh the culturing of naïve murine thin liver slices (250 µm) in conjunction with exposure to soluble egg antigens. This system has allowed us to Stanford University School of Medicine, Stanford, CA, United States identify the transcriptional events that contribute to the initiation of the Infection with Schistosoma haematobium, the cause of urogenital subsequent granulomatous response. Initially tissue, without parasite schistosomiasis, is a risk factor for bacterial urinary tract co-infection. This antigen exposure, was analysed for general histological changes, the co-infection worsens the sequelae of urogenital schistosomiasis, including presence of liver enzymes indicative of hepato-toxicity and finally RNA hematuria, dysuria, and risk of bladder cancer. Despite the impact of quality. All of these parameters indicated the fidelity of the tissue, these infections, it is unknown how co-infection by S. haematobium and and the sterility over 48 hour period were maintained. Next to build bacterial uropathogens impairs host clearance of bacterial UTI. Many on these research tools, we have employed microarray analysis of helminth infections, including schistosomiasis, induce host leukocytes the tissue with and without parasite egg antigen, in order to allow to secrete IL-4. It is unknown whether IL-4 impairs bacterial clearance in us to follow the dynamics of antigen presentation, inflammation and schistosome-bacterial co-infections. Another ill-defined but key facet of general hepatotoxicity, which represent the initial phases that will lead anti-bacterial immunity is the role of iNKT cells. To study the mechanisms to pathology. Findings from this ex vivo approach, are currently being of S. haematobium-bacterial uropathogen co-infections, we combined integrated with data from our previously in vivo whole organ studies with the first tractable model of urogenital schistosomiasis with an established Schistosoma japonicum. We eventually aim to identify the contribution of mouse model of bacterial UTI. This model recapitulates human co- hepatic and systemic immune cell types in the host transcriptional response infection, since a single bladder exposure to S. haematobium eggs triggers to egg deposition and the resulting granuloma formation in host liver. IL-4 production and renders a mouse strain susceptible to bacterial UTI when it otherwise is resistant (BALB/c). During co-infection, bladders are infiltrated by fewer iNKT cells than during bacterial UTI alone. Moreover, 1000 co-infection results in lower CD1d expression in bladder dendritic cells and lower levels of IFN- in bladder iNKT cells on a per-cell basis. We have SCHISTOSOMIASIS JAPONICA DURING PREGNANCY IS γ found that three distinct conditions can restore the baseline resistance of ASSOCIATED WITH ELEVATED ENDOTOXIN LEVELS IN BALB/c mice to bacterial UTI despite prior exposure to S. haematobium MATERNAL AND PLACENTAL COMPARTMENTS eggs: 1) antibody neutralization of IL-4, 2) genetic deficiency of IL-4 Emily A. McDonald1, Sunthorn Pond-Tor1, Blanca Jarilla2, receptor-α signaling, and 3) exogenous glycolipid antigen-induced Marianne Saglib2, Analisa Gonzal2, Remigio Olveda2, Luz Acosta2, activation of iNKT cells. We hypothesize that S. haematobium egg-induced Fusun Gundogan3, Lisa M. Ganley-Leal1, Jonathan D. Kurtis1, IL-4 reduces CD1d expression by antigen-presenting cells, which dampens Jennifer F. Friedman1 iNKT cell-derived, IFN-γ-mediated clearance of bacteriuria. Continuing work will test this hypothesis, and may enable iNKT cell-based therapies as 1Rhode Island Hospital, Providence, RI, United States, 2Research Institute alternatives to antibiotic treatment of UTI more broadly. of Tropical Medicine, Manila, Philippines, 3Women and Infants Hospital, Providence, RI, United States 1002 Schistosomiasis affects approximately 40 million women of reproductive age, and chronic infection has been linked to elevated levels of endotoxin SCHISTOSOMIASIS JAPONICA SOLUBLE EGG ANTIGENS in circulation. Whether this is also true in pregnancy complicated with INTERFERE WITH DIFFERENTIATION AND INVASION OF schistosomiasis has not been evaluated. In this study, we measured PLACENTAL TROPHOBLAST CELLS endotoxin levels in maternal peripheral (32 wks gestation), placental and newborn plasma collected from a cohort of 133 women in Leyte, The Emily A. McDonald1, Fusun Gundogan2, Jennifer F. Friedman1, Philippines. Birth outcomes, cord blood, placental biopsy and placental Jonathan D. Kurtis1 blood were collected at delivery; endotoxin levels were measured in 1Rhode Island Hospital, Providence, RI, United States, 2Women and Infants all plasma samples. Placental biopsies were evaluated for a number of Hospital, Providence, RI, United States histopathological outcomes related to placental inflammation. After Schistosomiasis represents a significant disease burden in endemic adjusting for confounders, endotoxin levels in pregnant women with regions. We have previously shown that schistosomiasis during pregnancy schistosomiasis were higher in the maternal and placental plasma results in a pro-inflammatory cytokine response detectable in maternal, than in uninfected women (1.3-fold, P = 0.03 and 2.4-fold, P < 0.001, placental, and cord blood as well as increased pathological signs of respectively). Premature birth and acute chorioamnionitis were associated placental inflammation. Our previous data suggest this response is at least with elevated levels of endotoxin in the placental plasma (2.5-fold partly due to altered cytokine production by the trophoblast cells of the higher endotoxin in premature births, P = 0.01, 2.0-fold in acute placenta. In addition to immune balance, trophoblasts are responsible chorioamnionitis, P = 0.04). A host of pro-inflammatory cytokines such for the majority of placental functions, including invasion of the uterine as IL-6 (7.1-fold, P < 0.001), TNF-α (6.1-fold, P <0.001), IFN-γ (1.8-fold, lining and hormone production for the maintenance of pregnancy. P 0.05), IL-1 (14.7-fold, P <0.001), and CRP (2.8-fold, P <0.001) were Herein, we have expanded our previous data to examine the effect of elevated in maternal plasma among women with endotoxin levels in the schistosome soluble egg antigens (SEA) on specific aspects of trophoblast highest tertile of the distribution. Additionally, some anti-inflammatory function, including differentiation, invasion and hormone production. cytokines including IL-10 (2.1-fold, P <0.001), IL-5 (2.2-fold, P =0.01), and Primary cytotrophoblasts were collected at term from uninfected North IL-13 (2.1-fold, P <0.001) were elevated in the placental plasma of these American placentas and placed in culture for 5d, during which time subjects. We have previously shown that schistosomiasis during pregnancy they spontaneously differentiate to syncytiotrophoblast, the cell layer can elicit a pro-inflammatory cytokine response in placental, maternal and responsible for nutrient/gas/waste exchange and hormone production. cord blood. Herein, we report for the first time that S. japonicum infection Real-time qPCR for syncytin, a marker of trophoblast differentiation, showed a decrease in mRNA levels in cells exposed to SEA for the final astmh.org 305 24h of the culture period. Progesterone production in this model system sequencing of an is-ncRNA-specific library. A total of 1,159 novel is- also tended to be lower in trophoblasts exposed to SEA. In addition, ncRNAs, including antisense, intergenic, and intronic is-ncRNAs were we used the HTR8/SVneo cell line as a model of extravillous trophoblast identified. Bioinformatics analyses indicated that the intergenic is-ncRNAs with invasive characteristics. Treatment of HTR8 cells with SEA for 24h were the least conserved among eight different Plasmodium species, and resulted in a significant drop in the number of cells that invaded through a antisense is-ncRNAs were more conserved than their sense counterparts. matrigel-coated membrane. Interestingly, there was no difference in either Thirty-six novel sno/scaRNAs were predicted, seven potential novel classes model system in the degree of TUNEL staining, suggesting SEA does not of is-ncRNAs were discovered by clustering analysis, and two novel cause apoptosis in trophoblast cells. These data suggest that SEA interferes internal motifs of intergenic is-ncRNAs were identified. The expression with the differentiation and function of trophoblast cells, and thus may of selected novel is-ncRNAs was confirmed by RT-PCR and northern predispose affected pregnancies to poor placentation and consequent blotting assays. An obvious difference in the novel is-ncRNA expression birth outcomes. profiles of the early and late intraerythrocytic developmental stages of the parasite was observed by Realtime PCR, suggesting the expression 1003 of the novel is-ncRNAs is regulated tightly in the parasite. Many of the novel is-ncRNAs showed a higher expression
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