In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: The application of the enzyme-labeled antigen method.
In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: The application of the enzyme-labeled antigen method.
复制标题
牙周炎中产生与牙龈卟啉单胞菌反应的抗体的浆细胞的原位可视化：酶标记抗原方法的应用。
DOI：
10.1111/mom.12052
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Tsutsumi Y.
中科院分区：
文献类型：
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作者：
Mizutani Y;Tsuge S;Takeda H;Hasegawa Y;Shiogama K;Onouchi T;Inada KI;Sawasaki T;Tsutsumi Y.
牙龈卟啉单胞菌是一种重要的牙周致病菌。组织学上，牙周炎的牙龈组织中有致密的浆细胞浸润。然而，免疫细胞分泌的抗体识别的抗原仍然未知。应用酶标抗原法检测牙周炎牙龈组织中产生牙龈卟啉单胞菌特异性抗体的浆细胞。使用小麦胚芽提取物通过无细胞蛋白质合成系统制备N末端生物素化的牙龈卟啉单胞菌抗原、Ag 53和四个牙龈卟啉菌蛋白酶结构域（Arg‐pro、Arg‐hgp、Lys‐pro和Lys‐hgp）。以这5种标记蛋白为探针，对20例牙周炎病变进行了检测。用AlphaScreen方法，在11份（55%）血清样本和17份（85%）组织提取物中检测到针对五种牙龈卟啉单胞菌抗原中任何一种的抗体。在牙龈组织的多聚甲醛固定冷冻切片上使用酶标记抗原法，在18份标本中的17份（94%）中用5种抗原中的任何一种标记浆细胞，其中检测到可评价的浆细胞。牙周炎的阳性率显着高于之前在根性囊肿中发现的阳性率（AlphaScreen方法在血清中为20%，组织提取物中为33%，酶标记抗原方法为25%）。我们的研究结果直接表明，牙龈卟啉单胞菌抗体反应是局部产生的牙龈病变，并对牙龈卟啉单胞菌的炎症反应参与牙周炎。
Porphyromonas gingivalisis a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme‐labeled antigen method was applied to detecting plasma cells producingP. gingivalis‐specific antibodies in biopsied gingival tissue of periodontitis. N‐terminally biotinylatedP. gingivalisantigens, Ag53 and four gingipain domains (Arg‐pro, Arg‐hgp, Lys‐pro and Lys‐hgp) were prepared by the cell‐free protein synthesis system using wheatgerm extract. With these five labeled proteins as probes, 20 lesions of periodontitis were evaluated. With the AlphaScreen method, antibodies against any one of the fiveP. gingivalisantigens were detected in 11 (55%) serum samples and 17 (85%) tissue extracts. Using the enzyme‐labeled antigen method on paraformaldehyde‐fixed frozen sections of gingival tissue, plasma cells were labeled with any one of the five antigens in 17 (94%) of 18 specimens, in which evaluable plasma cells were detected. The positivity rates in periodontitis were significantly higher than those found previously in radicular cysts (20% in sera and 33% in tissue extracts with the AlphaScreen method, and 25% with the enzyme‐labeled antigen method). Our findings directly indicate that antibodies reactive toP. gingivalisare locally produced in the gingival lesions, and that inflammatory reactions againstP. gingivalisare involved in periodontitis.