MAC-1 LEUKOCYTE GLYCOPROTEINS IN PERIODONTITIS

中性粒细胞/单核细胞趋化功能紊乱一直是 表现为K局限性青少年牙周炎(LJP)或全身性 青少年牙周炎(GJP)综合征，但分子致病 这些细胞异常的机制还不完全清楚。 已澄清。最近的研究发现了一种新发现的基因 以反复软组织感染为特征的疾病(包括 严重的青春期前全身性牙周炎)，以及严重的 中性粒细胞/单核细胞趋化及其他黏附依赖性细胞功能 这与一种“粘合剂”家族的缺陷有因果关系。 糖蛋白(Mac-1[IC3b受体]、LFA-1和P150、95)在 白细胞表面。拟议的研究将评估：(1)潜力 鸡肺炎Mac-1糖蛋白表达/功能的内在异常 GJF白细胞的LJP，以及(2)微生物可能的病理效应 Mac-1蛋白上的产物。黏附依赖的PMN/单核细胞功能 (趋化、聚集、IC3b吞噬或细胞毒性) 在趋化条件下进行评估，旨在最大限度地提高Mac-1和 P150，95表面表达。这些研究将与量化相关 用特异性单抗评估表面蛋白的表达 免疫荧光流式细胞术和125I中的抗体 免疫沉淀技术。因为我们的初步研究表明 抑制刺激的Mac-1表达和LJP中性粒细胞的过度黏附， 将使用其他技术来量化细胞内池 细胞组分中的Mac-1蛋白(刀豆球蛋白A-免疫印迹 技术)或在EBV转化中生物合成这些分子 淋巴细胞(35S蛋氨酸标记)。抗Mac-1单抗的应用 将尝试在LJP中识别受影响的患者或杂合子 可能有助于阐明遗传病遗传传播的家系 疾病。“牙周病”可能的继发性影响 还将评估微生物对Mac-1糖蛋白表达的影响； 将正常的中性粒细胞/单核细胞与培养滤液或超声波孵育。 放线菌伴生放线杆菌、类杆菌和Capnocytophaga sp. 在趋化性和其他黏附依赖功能分析中，以及 同时评估关于…的表达、功能或诱导 Mac-1。我们的研究应该通过以下方式扩展之前的实验观察： (1)为趋化性受损等提供分子基础 LJP家系中的白细胞功能异常(2)勾画 内在(遗传)和/或继发性病理机制和(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。

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.