牙周炎患者血清与龈沟液特异抗体水平及治疗前后变化

用酶联免疫吸附法检测抗牙类杆菌（Ｂｇ）抗体ＩｇＧ，纵向观察了２０例牙周炎患者治疗前、治疗后１和７个月其血清和龈沟液（ＧＣＦ）特异抗体的变化，以及血清与ＧＣＦ抗体水平的关系。结果显示，治疗前ＧＣＦ与血清（Ｓｒ）抗Ｂｇ抗体水平比值（ＧＣＦ与Ｓ?比值）显著１，且低于健康对照组；而治疗后１个月该比值明显增加至大于１。治疗后血清抗体水平持续下降，而ＧＣＦ抗体水平治疗１个月后明显上升，７个月下降。ＧＣＦ抗体水     

侵袭性牙周炎患者血清抗牙龈卟啉菌表面抗原抗体滴度和亲和力的研究

临床上通过检测抗牙龈卟啉菌(Porphyromonas gingivalis,Pg)表面抗原脂多糖(lispopolysaccharide，LPS)的血清抗体滴度和亲和力来评估体液免疫反应在侵袭性牙周炎(aggressive periodontilis，AgP)中抵御Pg的作用。研究表明，在评估体液免疫反应在抵御牙周致病菌的作用时，抗体滴度更有价值。然而，抗体水平和亲和力之间的关系目前尚存有争议。此研究旨在探讨AgP患者血清中抗Pg表面抗原的IgG抗体的滴度和抗体亲和力之间的关系。     

牙周病患者血清及龈沟液特异抗体水平与龈下细菌量的关系

采用ＥＬＩＳＡ检测３９名不同类型牙周病患者的血清及其１５６颗患牙的龈沟液（ＧＣＦ）中抗牙龈叶琳菌（Ｐｇ）抗体（ＩｇＧ）水平，并用间接免疫荧光法检测龈下菌斑中的Ｐｇ，观察Ｐｇ检出量、血清（Ｓｒ）和ＧＣＦ抗体水平、二者抗体水平比值（ＧＣＦ／Ｓｒ比值）以及与临床指数间的关系。结果显示，Ｐｇ检出量与疾病类型、临床指数呈正相关，与ＧＣＦ抗Ｐｇ抗体及ＧＣＦ／Ｓｒ比值呈负相关。初步证实龈下菌斑中细菌量增多，会使ＧＣＦ中相应抗体水平下降，从而使ＧＣＦ／Ｓｒ比值降低。ＧＣＦ抗Ｐｇ抗体水平、ＧＣＦ／Ｓｒ比值及龈下菌斑中Ｐｇ检出量结合起来，有可能成为牙周病活动期的诊断指标。     

牙周炎患者拔牙前后血清抗牙龈卟啉菌抗体水平的变化

用酶联免疫吸附测试法检测了７例成人牙周炎（ＡＰ）患者全口拔牙前及拔牙后１、２、３、１２及１８个月的血清抗牙龈卟啉菌（Ｐｇ）抗体水平的变化，并与６名牙周健康成人血清进行对照。结果表明，７例ＡＰ患者全口拔牙后血清抗ＰｇＩｇＧ抗体均逐月下降且明显低于拔牙前水平，但在拔牙后１８个月时仍明显高于对照组水平。血清抗体水平基本以相近似的速率下降，其中拔牙后１个月时的下降率较低。其下降率与牙周病各临床指数之间未显示明显的相关性。拔牙前的抗体水平与全口存留牙的深牙周袋率之间呈正相关趋势。本研究结果提示，在Ｐｇ感染去除后，机体对Ｐｇ抗原所产生的抗体逐渐下降，以牙周病患者血清抗ＰＧＩｇＧ水平作为该菌感染存在的诊断指标之一，值得进一步研究。     

牙龈卟啉菌蛋白酶的研究进展

革兰氏阴性厌氧菌牙龈卟啉菌和牙周炎病因密切相关,生物化学和分子生物学的进展表明牙龈卟啉菌可合成多种蛋白酶,蛋白酶是细菌重要的毒力因子,具有复杂的生理功能。本文简述了牙龈卟啉菌蛋白酶的生物学作用及其在牙周炎组织破坏中的影响。     

牙龈卟啉菌的抗原学研究新进展

牙龈卟啉菌作为牙周炎的一个重要致病菌受到各国牙周病学家的重视。本文综述了Ｐ．ｇ各种菌体抗原的研究进展，进而阐述了人们对其各种抗原成份的看法。     

牙龈卟啉菌、中间普氏菌的分离、培养和鉴定

目的：采用厌氧培养和鉴定技术，分离牙周炎患者龈下菌斑中的牙龈卟啉菌和中间普氏菌。方法：采集牙周炎患者的龈下菌斑，厌氧培养，分离产黑色素菌。经长波长紫外灯观察，各种生化分析和间接免疫荧光染色，分离和鉴定牙龈卟啉菌，中间普氏菌。结果：在受检的33例牙周炎患者中，24例患者检出了牙龈卟啉菌，总检出率为72．77％，共分离了79株。18例患者检出了中间普氏菌，总检出率为54．54％。共分离了32株。结论：厌氧培养，生化反应鉴定技术的发展，使牙龈卟啉菌和中间普氏菌的分离与培养准确可靠，为今后在分子水平上了解各菌株的致病机理打下了基础。     

牙龈素在牙龈卟啉单胞菌生长及牙周致病过程中的作用

牙龈素是慢性牙周炎的重要致病菌---牙龈卟啉单胞菌的最重要毒力因子。作为一组半胱氨酸蛋白酶,牙龈素能结合并降解多种宿主蛋白质。本文总结了目前对牙龈素的结构和功能的研究,主要从牙龈素对牙龈卟啉单胞菌生长代谢、对牙周组织的致病作用及宿主免疫的影响三个方面阐述。     

牙周炎患者基础治疗后牙龈卟啉单胞菌的定植研究

目的 通过对牙周炎患者龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalu,Pg)的检测,探讨慢性牙周炎(chronic periodontitis,CP)和侵袭性牙周炎(aggressive periodontitis,AgP)患者牙周基础治疗后Pg的定植规律.方法 选取90例CP患者和90例AgP患者,在牙周基础治疗前、治疗后6周、12周共采集龈下菌斑样本1620个,运用AmpliFluor终末点定量聚合酶链反应方法 检测Pg含量.结果 治疗后6周CP和AgP组Pg活动位点分别为61(22.6%)和66(24.4%)个,两者差异无统计学意义(P>0.05);治疗后6周Pg活动位点在治疗前检测的牙周临床指数高于Pg静止位点.治疗后12周两组Pg活动位点分别为96(35.6%)和18(6.7%)个,差异有统计学意义(P<0.05);治疗后12周Pg活动位点在治疗后6周时检测的牙周临床指数高于Pg静止位点.结论 在牙周基础治疗后6周时,CP和AgP患者Pg定植均已开始,仉是两组定植规律存在一定差异.在牙周基础治疗后,治疗前牙周组织炎性反应严重的龈下位点Pg易于定植.     

Fusobacterium nucleatum impairs serum binding to Porphyromonas gingivalis biofilm

Mouse immune sera obtained by immunization with Fusobacterium nucleatum and then Porphyromonas gingivalis demonstrated an impaired binding capacity to P. gingivalis-biofilm and lower avidity to P. gingivalis when compared with sera obtained from mice immunized with P. gingivalis alone.     

Serum antibody reactivity against recombinant PrtC of Porphyromonas gingivalis following periodontal therapy

OBJECTIVES: In 34 patients with chronic periodontitis, the presence of IgA, IgG, and IgG subclass serum antibodies against recombinant PrtC (rPrtC) of Porphyromonas gingivalis was assessed by immunoblot analysis 24 months after therapy. METHODS: rPrtC was produced from P. gingivalis ATTC 33277 using the plasmid pGEX-2T. In addition, intraoral colonization with P. gingivalis was detected by PCR in subgingival plaque and swab samples from buccal mucosae, tonsils and tongue at baseline, 10 d, and 3, 6, 9, 12, 18, and 24 months. RESULTS: All patients were found to harbor P. gingivalis in the oral cavity at least once during the observation period. The identified antibody responses against the rPrtC of P. gingivalis were IgA (97%, i.e. 33/34 patients) and IgG (100%, i.e. 34/34), with an IgG subclass distribution of IgG2 (65%, i.e. 22/34 patients) > IgG3 (47%, i.e. 16/34) > IgG1 (38%, i.e. 13/34) > IgG4 (29%, i.e. 10/34). Anti-rPrtC IgA and IgG antibody reactivity was found in all but one patients (anti-rPrtC IgA negative), who tested negative for P. gingivalis at all of the assessed intraoral sites for at least 6 months before sera collection. There was no association between IgG subclass reactivity against the rPrtC of P. gingivalis and progression of periodontal attachment loss. CONCLUSION: The results indicated that anti-rPrtC IgA and IgG antibodies may serve as an indicator for past or present intraoral colonization with P. gingivalis.     

Increased titre and avidity of IgG antibodies to Porphyromonas gingivalis whole cells and a cell surface protein in subjects with adult periodontitis

The titre and avidity of IgG antibodies to Porphyromonas gingivalis whole cells and a 47 kDa cell surface protein were determined in scrum samples taken from 20 subjects with adult periodontitis and 20 controls, matched for age, gender, ethnic origin and oral hygiene status. Antibody litres were measured by ELISA and antibody avidity was determined by a chaotrope-dissociation ELISA. Avidity was defined as the molarity of chaotrope required to reduce absorbance by 50% (ID₅₀). The mean IgG antibody log titre to whole cells (8. 29 vs. 6. 92; p<0.01) and to the 47 kDa antigen (7. 61 vs. 6. 77; p<0.05) were higher in cases than in controls. Mean IgG antibody avidity to whole cells (4. 59 vs. 2. 47; p<0.001) and to the surface protein (2. 54 vs. 1. 67; p<0.001) were also higher in cases than in controls. In cases, IgG antibody titre was highly correlated with avidity for both whole cells (r=0.878; p= <0.001) and the 47 kDa protein (r=0.683; p<0.001). There was a weaker positive correlation between the titre and the avidity of antibody to whole cells (r=0.591; p<0.01) in the control population but antibody titre and avidity for the 47 kDa sonicate antigen were not correlated in the controls (r=0.104). We conclude that many patients with adult periodontitis have effective humoral immunity to P. gingivalis. However, in up to half the patients with adult periodontitis, antibody titres and avidities were low and similar to control values, indicating either susceptibility due to poor host response or that disease is not associated with this particular pathogen.     

龈下菌斑中牙龈卟啉单胞菌rgpB基因多态性研究

目的 探讨不同rgpB基因型牙龈卟啉单胞菌在慢性牙周炎发生发展中所起的作用。方法 选择不同牙周状态下的龈下菌斑样本104个，设计引物对牙龈卟啉单胞菌rgpB催化域基因进行扩增，用限制性片段长度多态性分析的方法将牙龈卟啉单胞菌分为4个基因型。结果 慢性牙周炎患者病变重部位rgpB-cdⅣ型检出率最高为52．78％，与Ⅰ、Ⅲ型比较差异有统计学意义（P〈0．05、P〈0．01），病变轻部位rgpB-cdⅡ型检出率为75．86％，显著高于其他型（P〈0．01）。结论rgpB-cdⅣ型可能与慢性牙周炎的关系最密切，在致病过程中起主要作用，而rgpB-cdⅡ型可能与慢性牙周炎无关，为健康人群或牙周健康部位的定居菌。     

Serum IgG antibody response to Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis: implications for periodontal diagnosis

Abstract. The relationship of the serum antibody titer and avidity to the putative periodontal pathogens Actinobacillus actinomycetemcomitans (Aa) strains Y4 and 29523 and Porphyromonas gingivalis (Pg) strain 381 were examined in relation to clinical parameters in 26 gingivitis and 28 periodontitis patients. The relationship of antibody titer and avidity to infection with the homologous organism was also examined in a subset of 30 patients. Antibody titer was determined by an enzyme-linked immunosorbent assay, and antibody avidity was assessed using a dissociation assay. Considering all patients, there was a significant negative correlation between mean probing depth and antibody titer (r=-0.28) and avidity (r=-0.28) to Aa Y4. There was a significant positive correlation of probing depth and antibody titer (r=0.46) and avidity (r=0.46) to Pg. The correlation of antibody titer and avidity to Aa and infection with Aa Y4 (r=-0.32, r=-0.21) and Aa 29523 (r=-0.35, r=-0.39) was negative, while the correlations of titer and avidity to Pg and presence of the organisms was strongly positive (r=0.40, r=0.35). These data indicate that the relationship of serum antibody titer and avidity to clinical parameters of periodontal disease severity and the level of infection with the homologous organism appears to be different for Aa and Pg. The development of an antibody response to Aa appears to protect the individual from infection with the organism. In contrast, the development of an antibody response to Pg was not able to eliminate the infection. These results should be considered when developing a diagnostic strategy for periodontal disease utilizing the humoral immune response.     

高脂血症抑制载脂蛋白E基因敲除小鼠对牙龈卟啉单胞菌炎症反应的影响

目的:研究高脂血症在ApoE基因敲除(ApoE-/-)小鼠对牙龈卟啉单胞菌(P. gingivalis)炎症反应过程中的影响及其机制。方法:利用ApoE-/-小鼠建立长时间高脂血症模型,腹腔注射活P. gingivalis以建立腹膜炎症模型,倍比稀释法分析腹腔内清除细菌能力,ELISA法检测血液中白细胞介素6(IL-6)和单核细胞趋化因子1(MCP-1)的分泌,实时定量PCR检测腹腔细胞IL-6和MCP-1的转录水平,采用SPSS13.0软件包进行统计学分析。结果:高脂血症显著降低了腹腔清除P. gingivalis的能力,ApoE-/-小鼠腹腔细胞中IL-6和MCP-1转录水平下降,释放入血液中的IL-6和MCP-1水平下降。结论:高脂血症导致宿主对P. gingivalis炎症反应不足,清除细菌能力下降,加速了牙周病的发展进程。     

牙龈卟啉单胞菌刺激树突状细胞激活炎症反应

[摘要] 目的 分析树突状细胞在牙龈卟啉单胞菌感染后的差异表达基因,探讨牙周炎的致病机制及为日后研究提供线索。方法 利用高通量测序技术来分析体外诱导分化的树突状细胞在牙龈卟啉单胞菌感染情况下的差异表达基因、基因集及相关功能。 结果 本实验总计发现7305个至少有两倍变化的差异表达基因。基因本体结果显示牙龈卟啉单胞菌感染后树突状细胞的生物过程、细胞成份、分子功能均发生了较大变化。基因集富集分析结果显示肿瘤坏死因子基因集、细胞因子及炎症反应基因集、白介素1及转录因子NF-kB家族基因集、干扰素信号基因集、B细胞受体信号基因集、T细胞受体信号基因集富集明显。结论 树突状细胞在牙龈卟啉单胞菌感染后会导致基因水平的变化,引起机体免疫反应及炎症反应的发生, CXCL及SOCS的水平及作用可作为日后研究的方向。     

Identification of the major surface protein antigens of Porphyromonas gingivalis using IgG antibody reactivity of periodontal case-control serum

The identity of the major surface antigens of Porphyromonas gingivalis was investigated. Outer membranes of P. gingivalis strains W83, W50, 381 and NCTC 11843 were prepared following inactivation of the trypsin-like enzyme activity. Three proteins, molecular weight 115, 55 and 40 kDa, were major components of the outer membranes of strains W83 and W50 and were also present in strains 381 and NCTC 11834. Two proteins, 55 and 47 kDa, were released from the cells during the sonication step of the outer membrane preparation procedure. Immunoblots using preparations of P. gingivalis W83 and serum from a case-control study of adult periodontitis demonstrated higher mean antibody reactivity in the case population to all the major proteins except for the 115 kDa outer membrane protein, which was recognized equally well by both populations. We conclude that the 55, 47 and 40 kDa proteins are important surface antigens of P. gingivalis. Characterization of the structure and function of these components should lead to an improved understanding of the host-parasite interactions in adult periodontitis.