TLR Mediated Trophoblast Injury and Poor Pregnancy Outcomes in S. japonica

TLR 介导的粳稻滋养层损伤和不良妊娠结局

• 批准号: 8196378
• 负责人: Emily A McDonald
• 金额: $ 4.9万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-05 至 2014-07-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196378
• 关键词: Affect; Aftercare; Allogenic; Animal Model; Antigens; Apoptosis; Apoptotic; Area; Attenuated; Biological Assay; Biological Models; Birth; Birth Weight; Blocking Antibodies; Blood; Blood Circulation; Caring; Cells; Comprehensive Health Care; Data; Developing Countries; Environment; Enzyme-Linked Immunosorbent Assay; Equilibrium; Evaluation; Female of child bearing age; Fetal Growth Retardation; Fetus; First Pregnancy Trimester; Funding; Genetic Models; Growth and Development function; Health; Human; Immune system; In Vitro; Individual; Infection; Inflammatory; Injury; Interferons; Interleukin-6; Knockout Mice; Life; Ligands; Low Birth Weight Infant; MAP Kinase Gene; Mediating; Modeling; Morbidity - disease rate; Mus; Newborn Infant; Outcome; Outcome Measure; Parasites; Philippines; Physiological; Pilot Projects; Placenta; Praziquantel; Pregnancy; Pregnancy Outcome; Pregnant Women; Production; Randomized Controlled Trials; Receptor Activation; Reporting; Risk; Rodent Model; Sampling; Schistosoma; Schistosoma japonicum; Schistosomiasis; Serum; Source; Surface; Syncytiotrophoblast; System; TLR2 gene; TNF gene; Time; Tissues; Toll-Like Receptor 2; Toll-like receptors; Treatment Efficacy; United States National Institutes of Health; Woman; Work; base; cytokine; cytotrophoblast; egg; immune activation; immunocytochemistry; in vivo; mortality; novel; offspring; research study; response; standard of care; trophoblast

DESCRIPTION (provided by applicant): Human schistosomiasis infects 200 million individuals, including 40 million women of child bearing age, and is a significant cause of morbidity and mortality in areas where the parasite is endemic. Treatment with praziquantel (PZQ) is the standard of care for many of those suffering from schistosome infections, however pregnant women have been systematically excluded from this treatment option. Although rodent models of schistosome infection during pregnancy clearly demonstrate adverse birth outcomes, there remains little examination of such a phenomenon in humans. Data from our pilot study suggests schistosome infection in humans can result in increased trophoblast TNF-a; production and apoptosis, as well as lower birth weights. This application is novel in its evaluation of the effect of schistosome-released soluble egg antigen (SEA) on human trophoblasts. We hypothesize that SEA will activate toll-like receptors (TLR)-2 and -4 on the surface of the human syncytiotrophoblast (STB), leading to induction of a pro-inflammatory cascade by this cell layer. We will utilize the most relevant model system available, primary trophoblasts isolated at term from women living in a region of The Philippines in which S. japonicum is endemic. Trophoblasts will be examined after differentiation, as the STB is the cell in direct contact with maternal blood, and thus most likely to be exposed to SEA in vivo. Specific Aim 1 will isolate term cytotrophoblasts from healthy, uninfected women. After in vitro syncytialization the cells will be exposed to SEA and TLR activation assessed through apoptosis rates, intracellular TNF-a; levels, cytokine production, MAPK and NF?B activation. Completion of Specific Aim 1 will allow us to report on specific effects of SEA on STBs in humans. Specific Aim 2 will isolate term cytotrophoblasts from infected women PZQ treatment given during the first trimester. These samples are being collected as part of an ongoing, U01 funded, randomized controlled trial evaluating PZQ treatment in the first trimester in women with schistosomiasis. STB will be cultured and TLR activation assessed as in Specific Aim 1. Despite PZQ treatment, the tissue egg burden does not immediately decrease, thus completion of Specific Aim 2 will identify effects of remaining schistosome eggs as well as PZQ treatment on the STB. Specific Aim 3 will utilize a murine model to determine the specific TLR(s) activated by SEA on trophoblast cells. Based on data in other physiological systems, we hypothesize that both TLR2 and -4 will be activated. This will be examined by trophoblast response to schistosome infection in genetic models deficient in either TLR2 or -4. Findings from Specific Aim 3 will advance mechanistic understanding in the field, allowing for better care of pregnant women with schistosomiasis. To our knowledge, this application will be the first to evaluate STBs for a specific response to antigens released into circulation during schistosome infection. This work is crucial in that it will allow for more comprehensive care of women and their offspring affected by schistosomiasis during pregnancy. PUBLIC HEALTH RELEVANCE: Approximately 40 million women of childbearing age suffer from schistosome infection globally at any given time. Multiple studies in rodent models, as well as a few reports in humans, suggest that schistosome infection results in poor pregnancy outcomes. The mechanisms of this phenomenon are unclear, and will be investigated in this application in order to enhance the treatment and health of pregnant women with schistosomiasis, as well as that of their affected offspring.

描述（申请人提供）：人类血吸虫病感染了2亿人，其中包括4000万育龄妇女，是寄生虫流行地区发病和死亡的重要原因。吡喹酮（PZQ）治疗是许多感染性疾病患者的标准治疗方法，但孕妇一直被系统地排除在这种治疗选择之外。虽然啮齿动物模型在怀孕期间的寄生虫感染清楚地表明不利的出生结果，仍然很少有这样的现象在人类中的检查。来自我们的初步研究的数据表明，人类中的溶酶体感染可导致滋养层TNF-α产生和细胞凋亡增加，以及出生体重降低。该应用在其评价溶酶体释放的可溶性卵抗原（SEA）对人滋养层细胞的作用方面是新颖的。我们假设SEA将激活人合体滋养层（STB）表面上的Toll样受体（TLR）-2和-4，导致该细胞层诱导促炎级联反应。我们将利用现有的最相关的模型系统，从生活在菲律宾一个地区的妇女中分离出足月的原代滋养细胞，其中S。日本血吸虫是地方性的。将在分化后检查滋养层细胞，因为STB是与母体血液直接接触的细胞，因此最有可能在体内暴露于SEA。具体目标1将从健康、未感染的妇女中分离出足月细胞滋养层细胞。在体外合胞化后，将细胞暴露于SEA和TLR活化，通过凋亡率、细胞内TNF-α水平、细胞因子产生、MAPK和NF？B激活。具体目标1的完成将使我们能够报告SEA对人类STB的具体影响。具体目标2将从妊娠早期给予PZQ治疗的感染妇女中分离出足月细胞滋养层细胞。这些样本是作为正在进行的U 01资助的随机对照试验的一部分收集的，该试验评估了血吸虫病妇女在妊娠早期的PZQ治疗。将培养STB，并按照特定目标1评估TLR活化。尽管进行了PZQ处理，但组织虫卵负荷不会立即降低，因此完成特定目标2将确定剩余的卵裂体虫卵以及PZQ处理对STB的影响。Specific Aim 3将利用鼠模型来确定由SEA在滋养层细胞上激活的特异性TLR。基于其他生理系统中的数据，我们假设TLR 2和TLR 4都将被激活。这将通过滋养层对TLR 2或TLR 4缺陷的遗传模型中的溶酶体感染的反应来检查。具体目标3的发现将促进该领域的机制理解，从而更好地照顾血吸虫病孕妇。据我们所知，该应用程序将是第一个评估STB对单克隆抗体感染期间释放到循环中的抗原的特异性反应。这项工作至关重要，因为它将使怀孕期间感染血吸虫病的妇女及其子女得到更全面的护理。 公共卫生相关性：在任何特定时间，全球约有4000万育龄妇女遭受艾滋病毒感染。在啮齿动物模型中的多项研究以及在人类中的一些报告表明，寄生虫感染导致不良的妊娠结局。这种现象的机制尚不清楚，将在本申请中进行调查，以加强对血吸虫病孕妇及其受影响后代的治疗和健康。