Phagocyte Receptors for Lipid A

脂质 A 的吞噬细胞受体

• 批准号: 7214857
• 负责人: Douglas T Golenbock
• 金额: $ 46.92万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214857
• 关键词: Address; Adenovirus Vector; Appendix; Area; Bacterial Infections; Binding; Binding Proteins; Binding Sites; Biochemistry; Biological Testing; Breeding; CD14 gene; Caring; Cell Maturation; Cells; Complex; Confocal Microscopy; Critical Illness; Cytokine Activation; DNA; Data; Dendritic Cells; Dimerization; Endotoxins; Engineering; Event; Flagellin; Glycolipids; Goals; Host Defense; Human; Immunoelectron Microscopy; Immunoprecipitation; Inflammatory Response; Interleukin-1; Invasive; Label; Learning; Lesion; Leukocytes; Ligands; Lipid A; Lipopolysaccharides; Mediating; Membrane; Molecular; Molecular Genetics; Morbidity - disease rate; Mus; Mutagenesis; Names; Numbers; Pathway interactions; Patients; Phagocytes; Poly I-C; Predisposition; Process; Production; Productivity; Proteins; Reader; Reagent; Receptor Signaling; Recruitment Activity; Resistance; Role; Salmonella; Salmonella infections; Scanning; Sepsis; Septicemia; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Surface; Syndrome; TLR3 gene; TLR4 gene; TLR5 gene; TLR7 gene; TLR8 gene; Testing; Therapeutic; Thinking; Toll-like receptors; United States; Visual; Western Blotting; Work; adapter protein; cytokine; design; in vivo; knockout animal; lipopolysaccharide-binding protein; macrophage; microbial; monocyte; mortality; mutant; prevent; receptor; receptor function; resiquimod; response; size; toll-like receptor 4; transcription factor

DESCRIPTION (provided by applicant): Gram-negative bacterial septicemia remains an important cause of morbidity and mortality in the United States. Gram-negative sepsis begins when bacterial membranes shed endotoxin (lipopolysacchadde, LPS)and engage signaling receptors on the surface of phagocytes and other LPS-sensitive cells. Although additional receptor components may yet be discovered, the names of the basic components of the LPS response machinery are now known. These include LPS-binding protein (LBP), CD14, Toll-like receptor (TLR) 4, MD-2 and one or more "Toll, interleukin 1, resistance" (TIR)-domain containing adapter proteins. The hypothesis to be tested represents the current dogma: sepsis begins with LBP-mediated presentation of LPS to CD14. CD14 presents LPS to the TLR4/MD-2 complex. LPS binding to TLR4 results in receptor dimerization and recruitment of MyD88/Mal complexes resulting in the initiation of the NF-KB and IRF signal transduction pathways. These transcription factors drive cytokine production, resulting in septicemia. Many aspects of this dogma need considerable reassessment and refinement. For example, while preliminary data confirm that LPS directly binds to MD-2, and suggest that TLR4 is also bound, TLR4/MD-2 forms large multimers- much larger than dimmers- after being bound by LPS. In addition to MyD88 and Mal/TIRAP, at least two other adapter molecules, TRAM (which we recently discovered) and TRIF, are involved in the TLR4 pathway. We propose 4 specific aims: 1) To characterize the binding of LPS to TLR4 and MD-2. 2) To determine if the TLR4/MD-2 complex activates signal transduction by creating 'signa/osome' clusters. The aim is meant to precisely quantify receptor size and composition of the signalosome in LPS-stimulated cells. 3) To assess the roles of the five T/R domain containing adapter molecules in TLR4 signal transduction. This aim combines aspects of biochemistry, molecular genetics and confocal microscopy to define the role of adapters in LPS stimulation. And finally, 4) to characterize the role of TRAM, a newly discovered adapter molecule, in LPS signal transduction by generating and characterizing a mouse with a targeted deletion in TRAM. Cells from this mouse will be tested for responses to LPS and other microbial products. In addition, we will challenge this knockout animal with Salmonella to determine if and hot TRAM expression contributes to host defense.

描述（由申请方提供）：革兰氏阴性菌败血症仍然是美国发病率和死亡率的重要原因。革兰氏阴性脓毒症开始于细菌膜释放内毒素（脂多糖，LPS）并与吞噬细胞和其他LPS敏感细胞表面的信号受体结合。虽然可能还发现了其他受体成分，但LPS反应机制的基本成分的名称现在是已知的。这些包括LPS结合蛋白（LBP）、CD 14、Toll样受体（TLR）4、MD-2和一种或多种含有“Toll、白介素1抗性”（TIR）结构域的衔接蛋白。待检验的假设代表了当前的教条：败血症始于LBP介导的LPS向CD 14的递呈。CD 14将LPS呈递给TLR 4/MD-2复合物。LPS与TLR 4的结合导致受体二聚化和MyD 88/Mal复合物的募集，导致NF-κ B和IRF信号转导途径的启动。这些转录因子驱动细胞因子产生，导致败血症。这一教条的许多方面需要相当多的重新评估和完善。例如，虽然初步数据证实LPS直接结合MD-2，并表明TLR 4也结合，但TLR 4/MD-2在被LPS结合后形成大的多聚体-比二聚体大得多。除了MyD 88和Mal/TIRAP之外，至少还有另外两种衔接分子TRAM（我们最近发现的）和TRIF参与TLR 4通路。我们提出了4个具体目标：1）表征LPS与TLR 4和MD-2的结合。2)确定TLR 4/MD-2复合物是否通过产生“信号体”簇来激活信号转导。其目的是精确定量LPS刺激细胞中信号体的受体大小和组成。3)探讨5种含T/R结构域的接头分子在TLR 4信号转导中的作用。该目的结合生物化学、分子遗传学和共聚焦显微镜来定义适配器在LPS刺激中的作用。最后，4）通过产生和表征具有TRAM靶向缺失的小鼠来表征TRAM（一种新发现的衔接分子）在LPS信号转导中的作用。将检测该小鼠的细胞对LPS和其他微生物产物的反应。此外，我们将用沙门氏菌挑战这种敲除动物，以确定热TRAM表达是否有助于宿主防御。

项目成果

Cutting edge: cells that carry A null allele for toll-like receptor 2 are capable of responding to endotoxin.

• DOI: 10.4049/jimmunol.162.12.6971
• 发表时间: 1999-06
• 期刊: Journal of immunology
• 影响因子: 4.4
• 作者: H. Heine;C. Kirschning;E. Lien;Brian G. Monks;M. Rothe;D. Golenbock

Phagocytosis, innate immunity, and host-pathogen specificity.

吞噬作用，先天免疫和宿主 - 病原体特异性。

• DOI: 10.1084/jem.20031256
• 发表时间: 2004-01-05
• 期刊: JOURNAL OF EXPERIMENTAL MEDICINE
• 影响因子: 15.3
• 作者: Henneke, P;Golenbock, DT
• 通讯作者: Golenbock, DT

Atherogenic lipids and lipoproteins trigger CD36-TLR2-dependent apoptosis in macrophages undergoing endoplasmic reticulum stress.

• DOI: 10.1016/j.cmet.2010.09.010
• 发表时间: 2010-11-03
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Seimon TA;Nadolski MJ;Liao X;Magallon J;Nguyen M;Feric NT;Koschinsky ML;Harkewicz R;Witztum JL;Tsimikas S;Golenbock D;Moore KJ;Tabas I
• 通讯作者: Tabas I

Toll-like receptor-dependent discrimination of streptococci.

Toll 样受体依赖性链球菌鉴别。

• DOI: 10.1179/096805106x118762
• 发表时间: 2006
• 期刊: Journal of endotoxin research
• 作者: Santos-Sierra,Sandra;Golenbock,DouglasT;Henneke,Philipp
• 通讯作者: Henneke,Philipp

Membrane expression of soluble endotoxin-binding proteins permits lipopolysaccharide signaling in Chinese hamster ovary fibroblasts independently of CD14.

可溶性内毒素结合蛋白的膜表达允许中国仓鼠卵巢成纤维细胞中的脂多糖信号独立于 CD14。

• DOI: 10.1074/jbc.274.20.13993
• 发表时间: 1999
• 期刊: The Journal of biological chemistry
• 作者: Ingalls,RR;Monks,BG;Golenbock,DT
• 通讯作者: Golenbock,DT