Bcl-2 induced protection in severe sepsis

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展，但死亡率仍然很高。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的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，抗凋亡分子Bcl-2在T淋巴细胞、B淋巴细胞和肠上皮细胞中的过表达导致盲肠结扎穿孔（CLP）引起的小鼠脓毒症中的存活率增加。我们在初步实验中表明，Bcl-2在骨髓细胞中的过表达也提供了对CLP引起的脓毒症的保护，并且这种保护可以通过将Bcl-2骨髓细胞移植到缺乏成熟淋巴细胞（Rag-/-）的小鼠中来过继转移。 这些数据表明，淋巴细胞是不必要的Bcl-2过度表达，以提供保护，从CLP。该项目的第一个假设指出，Bcl-2的过表达诱导其产物在严重脓毒症中具有保护作用的基因的表达。额外的初步结果表明，正常小鼠的T淋巴细胞过继转移到腹膜时，可以提供保护，从CLP时，先前刺激的抗体CD 3或CD 90。因此，我们的第二个假设表明，用CD 3或CD 90抗体刺激T淋巴细胞导致细胞保护基因的表达，其产物提供了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)为了鉴定候选基因，其产物可能是 通过使用基因表达阵列比较对照和Bcl-2过表达的髓样细胞、T淋巴细胞和B淋巴细胞之间的基因表达，2)通过比较未刺激的T淋巴细胞和识别CD 3或CD 90的单克隆抗体刺激的淋巴细胞之间的基因表达，鉴定其产物可能在严重脓毒症中具有保护作用的候选基因。3)通过微阵列分析或蛋白质组学方法检测特定目标1和2中的候选保护基因在脓毒症中的功效。

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.