MAC-1 LEUKOCYTE GLYCOPROTEINS IN PERIODONTITIS

牙周炎中的 MAC-1 白细胞糖蛋白

• 批准号: 3221633
• 负责人: Donald C. Anderson
• 金额: $ 20.61万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-02-01 至 1990-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3221633
• 关键词: Actinobacillus; Bacteroides; Epstein Barr virus; cell aggregation; chemotaxis; child (0-11); cytotoxicity; human subject; leukocytes; membrane activity; periodontitis; phagocytosis; radiotracer

Disturbed PMN/monocyte chemotactic function has been consistently demonstrated in K localized juvenile periodontitis (LJP) or generalized juvenile periodontititis (GJP) syndromes, but molecular pathogenic mechanisms underlying these cellular abnormalities have not been fully elucidated. Recent studies have identified a newly recognized genetic disorder characterized by recurrent soft tissue infections (including severe prepubertal generalized periodontitis), and severe deficits of PMN/monocyte chemotaxis and other adherence-dependent cellular functions which are causally related to a deficiency of a family of "adhesive' glycoproteins (Mac-1 [IC3b receptor], LFA-1, and p150, 95) expressed on leukocyte surfaces. The proposed studies will evaluate: (1) potential intrinsic abnormalities of expression/function of Mac-1 glycoproteins of LJP of GJF leukocytes, and (2) possible pathologic effects of microbial products on Mac-1 proteins. Adherence-dependent PMN/monocyte functions (chemotaxis, aggregation, IC3b phagocytosis or cytotoxicity) will be assessed under chemotactic conditions designed to maximize Mac-1 and p150,95 surface expression. These studies will be related to quantitative assessments of surface protein expression employing specific monoclonal antibodies (MAb) in immunofluorescence flow cytometry and 125I immunoprecipitation techniques. Since our preliminary studies indicate impaired stimulated Mac-1 expression and hyperadherence of LJP PMNs, additional techniques will be used to quantitate intracellular pools of Mac-1 proteins in cellular fractions (conconavalin A - immunoblot technique) or biosynthesis of these molecules in EBV-transformed lymphocytes (35S methionine labeling). Applications of anti-Mac-1 MAbs will attempt to identify affected patients or heterozygotes among LJP kindreds which may allow an elucidation of the genetic transmission of disease. Possible secondary influences of "periodontopathic" microorganisms on Mac-1 glycoprotein expression will also be evaluated; normal PMN/monocytes will be incubated with culture filtrates or sonicates of Actinobacillus actinomycetemcomitans, Bacteroides and Capnocytophaga sp. in chemotaxis and other adherence-dependent functional assays, and concurrently assessed with respect to expression, function or induction of Mac-1. Our studies should extend previous experimental observations by: (1) providing a molecular basis for impaired chemotaxis and other abnormalities of leukocyte function among LJP kindreds, (2) delineating intrinsic (genetic) and/or secondary pathologic mechanisms and (3) providing a molecular marker for identification of affected individuals or heterozygotes for LJP or GJP traits.

PMN/单核细胞趋化功能一直受到干扰 K 表现为局限性幼年性牙周炎 (LJP) 或全身性牙周炎 幼年性牙周炎 (GJP) 综合征，但分子致病 这些细胞异常的机制尚未完全阐明 阐明了。 最近的研究发现了一种新认识的遗传基因 以反复软组织感染为特征的疾病（包括 严重的青春期前广泛性牙周炎）和严重的牙周缺陷 PMN/单核细胞趋化性和其他粘附依赖性细胞功能 这与“粘合剂”家族的缺乏有因果关系 糖蛋白（Mac-1 [IC3b 受体]、LFA-1 和 p150, 95）表达于 白细胞表面。 拟议的研究将评估：（1）潜力 Mac-1 糖蛋白表达/功能的内在异常 GJF 白细胞的 LJP，以及 (2) 微生物可能的病理作用 Mac-1 蛋白的产品。 粘附依赖性中性粒细胞/单核细胞功能 （趋化性、聚集性、IC3b 吞噬作用或细胞毒性） 在旨在最大化 Mac-1 的趋化条件下进行评估 p150,95表面表达。 这些研究将与定量相关 使用特定单克隆抗体评估表面蛋白表达 免疫荧光流式细胞术中的抗体 (MAb) 和 125I 免疫沉淀技术。 由于我们的初步研究表明 LJP PMN 的刺激 Mac-1 表达受损和过度粘附， 将使用其他技术来定量细胞内池 细胞组分中的 Mac-1 蛋白（伴刀豆球蛋白 A - 免疫印迹 技术）或这些分子在 EBV 转化中的生物合成 淋巴细胞（35S 蛋氨酸标记）。 抗 Mac-1 MAb 的应用 将尝试识别 LJP 中受影响的患者或杂合子 亲属关系可能有助于阐明基因传递 疾病。 “牙周病”可能产生的继发影响 微生物对 Mac-1 糖蛋白表达的影响也将得到评估； 正常 PMN/单核细胞将与培养物滤液或超声处理一起孵育 Actinobacillus actinomycetemcomitans、Bacteroides 和 Capnocytophaga sp。 在趋化性和其他粘附依赖性功能测定中，以及 同时评估表达、功能或诱导 麦克-1。 我们的研究应该通过以下方式扩展以前的实验观察： (1)为受损的趋化性和其他提供分子基础 LJP 亲属白细胞功能异常，(2) 描述 内在（遗传）和/或继发病理机制和（3） 提供分子标记以识别受影响的个体或 LJP 或 GJP 性状的杂合子。