TEMPORAL ANALYSIS OF PLATELETS AS CYTOTOXIC MEDIATORS IN SEPSIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 身体对感染性或非感染性侮辱的反应被称为全身炎症反应综合征(SIRS)。当已知这一过程是由感染性病原体引起时，术语脓毒症和全身炎症反应综合征是同义词。全身炎症反应综合征/败血症及其后遗症，包括急性肺损伤，仍然是儿童发病率和死亡率的主要原因。基于对SIRS分子过程的新认识，我们假设在SIRS的儿童中，发展为感染性休克、MODS和/或死亡与血小板颗粒酶的不同表达模式有关。 具体目标1：建立全身炎症反应综合征、感染性休克和/或多器官功能障碍综合征儿童和健康对照的基因组数据库。为了检验我们的中心假设，有必要建立一个包含22个个体的遗传和临床数据库，以说明宿主对SIRS反应的预期多样性。我们建议从CNMC PICU招募的22名SIRS患儿的同时采集的外周全血中分离细胞组分。将从每个SIRS参与者那里获得两份样本；一份将在SIRS开始时获得，另一份将在SIRS开始后48小时获得。还将从血样中分离出DNA。SIRS的参与者将从CNMC PICU招募。此外，还将从参加CNMC ED的20名健康正常对照儿童身上采集单个外周血液样本。 特异性目标2：对AIM 1中所有入选患者的血小板进行体外功能淋巴细胞杀伤试验。我们将根据建议的类别对患者进行分类：1)对照组，2)单纯SIRS，3)SIRS进展为感染性休克/MODS，4)SIRS进展为感染性休克/MODS和死亡。使用体外淋巴细胞杀伤试验，我们将确定每个参与者的血小板的细胞毒性程度。为了验证我们的中心假设，即从SIRS到感染性休克、MODS和/或死亡的进展取决于血小板颗粒酶的表达模式，我们将对每一种建议类别的细胞毒性进行比较。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The body's response to an infectious or non-infectious insult has been termed the systemic inflammatory response syndrome (SIRS). When this process is known to be caused by an infectious agent, the terms sepsis and SIRS are synonymous. SIRS/sepsis and its sequelae, including Acute Lung Injury, remain a leading cause of morbidity and mortality in children. Based on new insights into the molecular processes of SIRS, we hypothesize that in children with SIRS, the progression to septic shock, MODS, and/or death, is associated with the variable expression patterns of platelet granzymes. Specific Aim 1: Development of a genomic database of children with SIRS, septic shock, and/or MODS and healthy controls. In order to test our central hypothesis, it will be necessary to establish a genetic and clinical database of 22 individuals to account for the anticipated diversity of host responses to SIRS. We propose to isolate cell fractions from concurrent collections of peripheral whole blood from 22 children with SIRS recruited from the CNMC PICU. Two samples will be obtained from each SIRS participant; one will be obtained at the onset of SIRS and then the other sample 48 hours after that. DNA will also be isolated from the blood samples. SIRS participants will be recruited from the CNMC PICU. In addition, a single peripheral blood sample will be collected from 20 healthy normal control children enrolled in the CNMC ED. Specific Aim 2: Perform functional in vitro lymphocyte killing assays on platelets from all enrolled patients in Aim 1. We will classify our patients according to the proposed categories: 1) controls, 2) SIRS alone, 3) SIRS progressing to septic shock/MODS, and 4) SIRS progressing to septic shock/MODS and death. Using the in vitro lymphocyte killing assays, we will determine the degree of cytotoxicity of each participant's platelets. Cytotoxicity in each of the proposed categories will be compared in order to test our central hypothesis that the progression from SIRS to septic shock, MODS, and/or death is dependent on the expression patterns of platelet granzymes.