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)发生的最好的单一预测因子。脓毒症的高死亡率，加上最近临床试验令人失望的结果，表明持续需要新的方法来治疗这种疾病。最近的研究表明，在盲肠结扎穿孔(CLP)所致的小鼠脓毒症模型中，T淋巴细胞、B淋巴细胞和肠上皮细胞中抗凋亡分子Bcl2的过度表达可提高小鼠的存活率。我们在初步实验中表明，在CLP引起的脓毒症中，髓系细胞中过表达的Bcl-2也提供了保护，并且这种保护可以通过将Bcl-2髓系细胞移植到成熟淋巴细胞缺陷(RAG-/-)小鼠身上而过继转移。 这些数据表明，淋巴细胞并不是Bcl2过度表达所必需的，以提供对CLP的保护。该项目的第一个假设是，Bcl2的过度表达诱导了一个基因(S)的表达，该基因的产物(S)在严重脓毒症中具有保护作用。另外的初步结果表明，当过继转移到腹膜时，来自正常小鼠的T淋巴细胞在先前用CD3或CD90抗体刺激时，可以提供对CLP的保护。因此，我们的第二个假设认为，抗CD_3或CD_(90)抗体刺激T淋巴细胞可导致细胞保护基因的表达(S)及其产物(S)对严重脓毒症具有保护作用。我们将通过以下具体目标来研究这些假设。1)确定候选基因(S)，其产物(S)可能是 通过使用基因表达阵列比较对照组和过度表达Bcl2的髓系细胞、T淋巴细胞和B淋巴细胞的基因表达，以保护严重脓毒症。2)通过比较未刺激的T淋巴细胞和识别CD_3或CD_(90)的单抗刺激的T淋巴细胞的基因表达，寻找其产物(S)可能在严重脓毒症中起保护作用的候选基因(S)。3)检测通过基因芯片分析或通过蛋白质组学方法确定的特定AIMS 1和2的候选保护基因在脓毒症中的疗效。

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.