Bcl-2 induced protection in severe sepsis

Bcl-2 在严重脓毒症中诱导保护

基本信息

批准号：
7522786
负责人：
ROBERT K WINN
金额：
$ 111.47万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7522786
关键词：
Activated Lymphocyte Adult Adult Respiratory Distress Syndrome Antibodies Apoptotic B-Lymphocytes CD3 Antigens Candidate Disease Gene Caring Cause of Death Cells Clinical Clinical Trials Complication Coupled Critical Illness Data Development Disease Doctor of Philosophy Epithelial Cells Failure Gene Expression Genes Healthcare Inflammation Investigation Life Ligation Lung Lymphocyte Mature Lymphocyte Methods Microarray Analysis Monoclonal Antibodies Multiple Organ Failure Mus Myeloid Cells Newborn Respiratory Distress Syndrome Numbers Other Therapy Outcome Patients Peritoneal Peritoneum Pharmaceutical Preparations Proteins Proteomics Publications Puncture procedure Research Sepsis Sepsis Syndrome T-Lymphocyte Therapeutic Transplantation United States Viral Vector activated Protein C base improved mortality pre-clinical preclinical study programs research study septic

项目摘要

Sepsis and/or sepsis syndrome constitutes a major cause of death in critically ill patients and mortality remains high despite advances in clinical care. Sepsis is the best single predictor for development of the Adult Respiratory Distress Syndrome (ARDS). The high mortality in sepsis coupled with the disappointing results from recent clinical trials demonstrates a continuing need for new methods of treating this disease. Recent publications have shown that over-expression of the anti-apoptotic molecule Bcl-2 in T-lymphocytes, B-lymphocytes and in gut epithelial cells results in increased survival in mouse sepsis resulting from cecal ligation and puncture (CLP). We show in preliminary experiments that over-expression of Bcl-2 in myeloid cells also provides protection in sepsis resulting from CLP and that this protection can be adoptively transferred through transplanting Bcl-2 myeloid cells into mice deficient in mature lymphocytes (Rag-/-). These data suggest that lymphocytes are not necessary for Bcl-2 over-expression to provide protection from CLP. The first hypothesis of this project states that over-expression of Bcl-2 induces the expression of a gene(s) whose product(s) is protective in severe sepsis. Additional preliminary results show that T-lymphocytes from normal mice when adoptively transferred to the peritoneum can provide protection from CLP when previously stimulated with antibodies to either CD3 or CD90. Thus our second hypothesis states that stimulation of T-lymphocytes with antibodies to either CD3 or CD90 results in expression of cyto-protective gene(s) and their product(s) provides protection from severe sepsis. We will investigate these hypotheses through the following specific aims. 1) To identify candidate gene(s) whose product(s) may be protective in severe sepsis by comparing gene expression between controls and Bcl-2 over-expressing myeloid cells, T-lymphocytes and B-lymphocytes using gene expression arrays. 2) To identify candidate gene(s) whose product(s) may be protective in severe sepsis by comparing gene expression between unstimulated T-lymphocytes and lymphocytes stimulated by monoclonal antibodies that recognize CD3 or CD90. 3) To examine the efficacy in sepsis of candidate protective genes as determined by either microarray analysis or by the proteomics approach in specific aims 1 and 2.

脓毒症和/或脓毒症综合征是危重患者死亡的主要原因，尽管临床护理取得了进步，死亡率仍然很高。脓毒症是成人呼吸窘迫综合征 (ARDS) 发展的最佳单一预测因子。脓毒症的高死亡率加上最近临床试验令人失望的结果表明，持续需要治疗这种疾病的新方法。最近的出版物表明，T 淋巴细胞、B 淋巴细胞和肠上皮细胞中抗凋亡分子 Bcl-2 的过度表达可导致盲肠结扎穿刺 (CLP) 引起的小鼠败血症的存活率增加。我们在初步实验中表明，骨髓细胞中 Bcl-2 的过度表达也对 CLP 引起的败血症提供保护，并且这种保护可以通过将 Bcl-2 骨髓细胞移植到成熟淋巴细胞缺陷的小鼠中来过继转移（Rag-/- ）。这些数据表明，Bcl-2 过度表达并不需要淋巴细胞来提供针对 CLP 的保护。该项目的第一个假设指出，Bcl-2 的过度表达会诱导基因的表达，该基因的产物在严重败血症中具有保护作用。其他初步结果表明，当先前用 CD3 或 CD90 抗体刺激时，来自正常小鼠的 T 淋巴细胞在过继转移到腹膜时可以提供针对 CLP 的保护。因此，我们的第二个假设表明，用 CD3 或 CD90 抗体刺激 T 淋巴细胞会导致细胞保护基因及其产物的表达，从而提供针对严重脓毒症的保护。我们将通过以下具体目标来研究这些假设。 1) 鉴定候选基因，其产物可能是通过使用基因表达阵列比较对照和 Bcl-2 过表达骨髓细胞、T 淋巴细胞和 B 淋巴细胞之间的基因表达，对严重脓毒症具有保护作用。 2) 通过比较未刺激的 T 淋巴细胞和识别 CD3 或 CD90 的单克隆抗体刺激的淋巴细胞之间的基因表达，鉴定其产物可能对严重脓毒症具有保护作用的候选基因。 3) 通过微阵列分析或特定目标 1 和 2 中的蛋白质组学方法确定候选保护基因在脓毒症中的功效。