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    • 3. 发明申请
    • CHOLINE BINDING PROTEINS FOR ANTI-PNEUMOCOCCAL VACCINES
    • 用于抗肺气肿疫苗的胆固醇蛋白
    • WO1997041151A2
    • 1997-11-06
    • PCT/US1997007198
    • 1997-05-01
    • THE ROCKEFELLER UNIVERSITY
    • THE ROCKEFELLER UNIVERSITYMASURE, H., RobertROSENOW, Carsten, I.TUOMANEN, ElaineWIZEMAN, Theresa, M.
    • C07K14/315
    • C07K14/3156A61K38/00A61K39/00A61K48/00A61K2039/51
    • The invention relates to bacterial choline binding proteins (CBPs) which bind choline. Such proteins are particularly desirable for vaccines against appropriate strains of Gram positive bacteria, particularly streptococcus, and more particularly pneumococcus. Also provided are DNA sequences encoding the bacterial choline binding proteins or fragment thereof, antibodies to the bacterial choline binding proteins, pharmaceutical compositions comprising the bacterial choline binding proteins, antibodies to the bacterial choline binding proteins suitable for use in passive immunization, and small molecule inhibitors of choline binding protein mediated adhesion. Methods for diagnosing the presence of the bacterial choline binding protein, or of the bacteria, are also provided. In a specific embodiment, a streptococcal choline binding protein is an enolase, which demonstrates strong affinity for fibronectin.
    • 本发明涉及结合胆碱的细菌胆碱结合蛋白(CBP)。 对于针对革兰氏阳性菌,特别是链球菌,更特别是肺炎球菌的合适菌株的疫苗,这种蛋白质是特别需要的。 还提供了编码细菌胆碱结合蛋白或其片段的DNA序列,细菌胆碱结合蛋白的抗体,包含细菌胆碱结合蛋白的药物组合物,适用于被动免疫的细菌胆碱结合蛋白的抗体和小分子抑制剂 的胆碱结合蛋白介导的粘附。 还提供了用于诊断细菌胆碱结合蛋白或细菌存在的方法。 在具体实施方案中,链球菌胆碱结合蛋白是烯醇化酶,其对纤连蛋白表现出强亲和力。
    • 5. 发明申请
    • USE OF GRAM-POSITIVE BACTERIA TO EXPRESS RECOMBINANT PROTEINS
    • 使用GRAM阳性细菌来表达重组蛋白
    • WO1996040943A1
    • 1996-12-19
    • PCT/US1996009965
    • 1996-06-06
    • THE ROCKEFELLER UNIVERSITYDARZINS, AldisWHITEHEAD, StephenHRUBY, Dennis
    • THE ROCKEFELLER UNIVERSITY
    • C12N15/62
    • C07K14/31A61K38/00A61K39/00C07K14/315C07K2319/00C07K2319/02C07K2319/035C07K2319/23C07K2319/50C07K2319/705C07K2319/735C12N15/62C12N15/74
    • A novel system for cloning and expression of genes in gram-positive bacteria. The expression system is based on the finding that many gram-positive bacteria sort proteins to their cell surface through cis-acting N-terminal signal sequences and C-terminal anchor regions. In particular, the cell sorting signals of the streptococcal M6 protein, a well-known surface molecule, are used to construct a gram-positive expression system, designated SPEX (Streptococcal Protein Expression). Expression is achieved by cloning the gene of interest into an appropriate SPEX cassette which is then stably introduced into a bacterial host, such as the human commensal Streptococcus gordonii. Depending on the SPEX vector used, recombinant proteins can be anchored to the cell wall prior to release by specific endoproteolytic cleavage or secreted into the culture medium during bacterial growth. The use of host bacteria lacking extracellular proteases should protect secreted proteins from proteolytic degradation. Several expression vectors in this system also produce specifically-tagged recombinant proteins which allows for a one-step purification of the resulting product.
    • 用于克隆和表达革兰氏阳性菌中基因的新系统。 表达系统基于许多革兰氏阳性细菌通过顺式作用的N末端信号序列和C末端锚定区域将蛋白质分解到其细胞表面的发现。 特别地,使用众所周知的表面分子的链球菌M6蛋白的细胞分选信号来构建称为SPEX(链球菌蛋白表达)的革兰氏阳性表达系统。 通过将感兴趣的基因克隆到适当的SPEX盒中,然后将其稳定地导入细菌宿主例如人共生型链球菌(Streptococcus gordonii)中来实现表达。 根据所使用的SPEX载体,重组蛋白质可以在通过特异性内切蛋白酶切割释放之前锚定到细胞壁上,或在细菌生长过程中分泌到培养基中。 缺乏细胞外蛋白酶的宿主细菌的使用应保护分泌的蛋白质免于蛋白水解降解。 该系统中的几种表达载体也产生特异性标记的重组蛋白,其允许对所得产物进行一步纯化。
    • 9. 发明申请
    • MODULATORS OF PNEUMOCOCCAL ADHESION TO CELLULAR TARGETS INVOLVING THE PLATELET ACTIVATING FACTOR RECEPTOR, AND USES THEREOF
    • 涉及平板激活因子受体的细胞粘附的肺炎粘附调节剂及其用途
    • WO1995035112A2
    • 1995-12-28
    • PCT/US1995007687
    • 1995-06-19
    • THE ROCKEFELLER UNIVERSITYBETH ISRAEL HOSPITAL ASSOCIATION
    • THE ROCKEFELLER UNIVERSITYBETH ISRAEL HOSPITAL ASSOCIATIONTUOMANEN, Elaine, I.CUNDELL, Diana, R.GERARD, Norma, P.
    • A61K31/70
    • A61K31/7008A61K31/00A61K31/341A61K31/343A61K31/551A61K38/1793A61K39/3955A61K31/7016A61K2300/00
    • The present invention relates to compositions and methods for preventing pneumococcal infection. In particular, the invention relates to identification of the major reception for Streptococcus pneumoniae on activated human cells, and diagnostic and therapeutic conpositions and methods based thereon. In particular, the invention relates to the discovery that platelet activating factor (PAF) receptor is an adhesive ligand for pneumococcal adherence to activated lung epithelial and venous endothelial (i.e., host) cells. Accordingly, the present invention is directed to a method for preventing or treating an infection with Streptococcus pneumoniae by administering an antogonist of platelet activating factor receptor. The invention further relates to recognition that adherence to activated cells also involves a carbohydrate ligand found on such activated cells. Thus, a method for inhibiting pneumococcal adherence may further comprise administering an amount of carbohydrate containing an N-acetyl-D-glucosamine motif. It has been found that resting lung epithelial and venous endothelial cells bear two classes of receptors containing different carbohydrate motifs. Thus, the invention further provides for administering an amount of a second carbohydrate selected from the group consisting of a carbohydrate containing a disaccharide N-acetyl-D-galactosamine beta 1-4Gal motif, a disaccharide N-acetyl-D-galactosamine beta 1-3Gal motif, and a mixture thereof. In addition, the invention provides pharmaceutical compositions comprising such agents that inhibit binding of pneumococci to human cells. In a specific example, platelet activating factor receptor antagonists and disaccharides are shown to inhibit binding of pneumococci to activated lung epithelial cells and venous endothelial cells, as well as cells transfected with the platelet activating factor receptor, in vitro.
    • 本发明涉及预防肺炎球菌感染的组合物和方法。 特别地,本发明涉及鉴定活化的人细胞上的肺炎链球菌的主要接收,以及基于此的诊断和治疗结合及方法。 特别地,本发明涉及血小板活化因子(PAF)受体是用于肺炎球菌粘附于活化的肺上皮和静脉内皮(即宿主)细胞的粘合剂配体的发现。 因此,本发明涉及一种预防或治疗肺炎链球菌感染的方法,该方法是通过施用血小板激活因子受体的脱发子。 本发明还涉及认识到对活化细胞的依附性还涉及在这种活化细胞上发现的碳水化合物配体。 因此,抑制肺炎球菌粘附的方法还可以包括给予一定量的含有N-乙酰基-D-葡糖胺基序的碳水化合物。 已经发现,休息的肺上皮和静脉内皮细胞具有包含不同碳水化合物基序的两类受体。 因此,本发明进一步提供一定量的第二种碳水化合物,其选自含有二糖N-乙酰基-D-半乳糖胺β1-4Gal基序的碳水化合物,二糖N-乙酰基-D-半乳糖胺β1- 3Gal图案及其混合物。 此外,本发明提供了包含抑制肺炎球菌与人细胞结合的药剂的药物组合物。 在具体实例中,血小板激活因子受体拮抗剂和二糖在体外显示出抑制肺炎球菌与活化的肺上皮细胞和静脉内皮细胞以及转染血小板活化因子受体的细胞的结合。