粘性放线菌菌毛的研究进展

粘性放线菌具有两种菌毛：Ｉ型和Ⅱ型，其功能和抗原性均不相同，Ⅰ型菌毛介导粘放菌粘附在有唾液包被的羟基磷灰石表面，受体是富脯蛋白和富酪蛋白，故Ⅰ型菌毛是通过蛋白质－蛋白质立体化学作用的模式而促进细菌的附着。Ⅱ型菌毛具有一种对乳糖特异性敏感的植物凝集素作用，介导粘放菌与血链菌的种间凝集反应。醉语文不粘性放线菌菌毛的提取方法，菌毛的化学及氨基酸组成，亚单位结构，稳定性等研究结果作一概述，并且就近年对粘放     

粘性放线菌对牙面粘附机理的研究Ⅳ.粘性放线菌唾液蛋白受体的研究

采用 3H-标记的粘性放线菌不同菌株 ,对 7种唾液蛋白包被羟基磷灰石的粘附实验 ,来筛选菌毛 及菌毛 的唾液蛋白受体。结果发现 :富脯糖蛋白 (PRPG)、酸性富脯蛋白及富酪蛋白是粘性放线菌菌毛 的受体 ;PRPG、SIg A是菌毛 的受体 ;PRPG是两种菌毛共同的受体 ;同时还发现不同唾液蛋白成份对不同的菌株粘附影响有差异 ,新分离的粘性放线菌菌株的粘附能力比参考株强。     

粘性放线菌菌毛的研究进展

粘性放线菌具有两种菌毛:Ⅰ型和Ⅱ型,其功能和抗原性均不相同。Ⅰ型菌毛介导粘放菌粘附在有唾液包被的羟基磷灰石表面,受体是富脯蛋白和富酪蛋白,故Ⅰ型菌毛是通过蛋白质—蛋白质立体化学作用的模式而促进细菌的附着。Ⅱ型菌毛具有一种对乳糖特异性敏感的植物凝集素作用,介导粘放菌与血链菌的种间凝集反应。本文就粘性放线菌菌毛的提取方法,菌毛的化学及氨基酸组成,亚单位结构,稳定性等研究结果作一概述,并且就近年对粘放菌Ⅰ型和Ⅱ型菌毛粘附性能、粘附机制的研究进展进行综述。     

粘性放线菌对牙面粘附机理的研究：Ⅲ.粘性放线菌变异株的鉴…

利用粘性放线菌的变异株制备全菌抗血清，采用免疫吸附、疏酸铵沉淀及离子交换层析制备了抗菌毛Ⅰ、ⅡＩｇＧ，并用ＳＤＳ－ＰＡＧＥ电泳测定其分子量，ＥＬＩＳＡ证明其特异性，为确定菌毛Ⅰ、Ⅱ在粘附中的作用，筛选粘性放线菌的粘结素成份奠定基础。     

粘性放线菌对牙面粘附机理的研究──Ⅱ．与牙面粘附有关的粘性放线菌粘结素的研究

本实验通过采用粘性放线菌不同菌株（包括培养上清液蛋白）对唾液包被羟基磷灰石的粘附实验及抗菌毛Ⅰ，Ⅱ的ＩｇＧ的粘附抑制实验来判断粘性放线菌粘结素存在部位，并确定不同菌毛在粘附中的作用。结果发现：１．粘性放线菌的粘结素主要位于菌毛上，并进一步证实两种菌毛共同参与粘性放线菌对牙面的粘附；２．上清蛋白中不含粘结素成份，可能含有有利于细菌凝集的成份；３．粘性放线菌地方株与参考株的菌毛有相同的抗原决定簇，为预防粘性放线菌地方株的粘附提供了实验依据。     

粘性放线菌菌毛生物性能的研究Ⅰ.粘性放线菌菌毛的提取和鉴定

目的：本实验旨在建立一套提取和鉴定粘性放线菌菌毛的方法。方法：采用粘性放线菌变异株５５１９（１＋２－）和５９５１（１－２＋）分别提取Ⅰ型菌毛和Ⅱ型菌毛。首先用机械搅动法分离菌毛与菌体,进一步用凝胶过滤柱层析提取、纯化两种菌毛,并通过电镜观察和对流免疫电泳进行初步鉴定。结果：证实分别提取到Ⅰ型菌毛和Ⅱ型菌毛。结论：两种菌毛的提取为进一步分析研究它们的生化特征和生物活性奠定了基础。     

粘性放线菌菌毛生物性能的研究：Ⅰ.粘性放线菌菌毛？…

本实验旨在建立一套提取和鉴定粘性菌菌毛的方法。方法采用粘性放线菌变异株５５１９（１＾＋２＾－）和５９５１（＾－２＾＋）分别提取Ⅰ型菌毛和Ⅱ型菌毛。首先用机械搅动法分离菌毛与菌体,进一步用凝胶过滤柱层析提取、纯化两种菌毛,并通过电镜观察和对流免疫电泳进行初步鉴定。     

粘性放线菌与龋病关系的研究现状

粘性放线菌是牙面早期定居细菌之一，它对龋病及牙周病的发生起着重要作用。现在许多学者从不同角度对它进行了研究。本文就近年来对粘性放线菌的分类鉴定，粘性放线菌的粘附及与粘附有关唾液蛋白，产酸等方面研究作一综述。     

粘性放线菌菌毛生物性能的研究Ⅱ.粘性放线菌菌毛化学组成的分析

目的：了解采用机械搅拌法提取的粘性放线菌Ⅰ型、Ⅱ型两种菌毛的化学组成。方法：首先制备出抗Ⅰ型菌毛抗血清和抗Ⅱ型菌毛抗血清,鉴定两种菌毛纯化物纯度后,采用考马斯亮蓝法和蒽酮法分别测定两种菌毛中蛋白质和糖的含量,并进行氨基酸组成分析。结果：两种菌毛的主要成分是蛋白质,糖含量较低。两种菌毛均是天门冬氨酸、谷氨酸、丙氨酸、赖氨酸和甘氨酸含量较多。两种菌毛中,非极性氨基酸百分含量最高,碱性氨基酸百分含量最低。结论：本实验结果有助于了解两种菌毛的化学组成与其生物性能的关系。     

粘性放线菌菌毛生物性能的研究Ⅲ.粘性放线菌菌毛粘附与凝集活性的研究

目的：了解粘放菌两种菌毛在粘附牙面和与血链球菌３４凝集中的生物活性。方法：采用粘放菌Ⅰ型和Ⅱ型菌毛提取物及两种抗菌毛抗体,通过它们对粘放菌粘附唾液包被的羟磷灰石（ＳＨＡ）的抑制实验来判断两种菌毛的粘附活性,通过粘放菌与血链球菌３４的凝集实验及两种抗菌毛抗体对凝集反应的抑制实验来确定两种菌毛的凝集活性。结果：①粘放菌Ⅰ型和Ⅱ型菌毛提取物及两种抗菌毛抗体均能抑制粘放菌对ＳＨＡ的粘附;②只有Ⅱ型菌毛能间接引起肉眼可见的血链球菌３４的凝集反应,抗Ⅱ型菌毛抗体能抑制凝集反应,抗Ⅰ型菌毛抗体无此抑制作用。结论：Ⅰ型和Ⅱ型菌毛均有粘附性能;Ⅱ型菌毛有凝集性能,Ⅰ型菌毛无凝集性能;本课题所采用的菌毛分离法未破坏菌毛的生物活性。     

粘性放线菌对牙面粘附机理的研究──Ⅰ．实验性全唾液获得性膜蛋白组份的分离纯化

采用羟基磷灰石柱层析形成实验性获得性膜的体外模型，研究全唾液获得性膜蛋白的组成成份。结合离子交换及分子筛层析分离获得８种获得性膜的蛋白成份。根据层析用谱、等电点、氨基酸组成及免疫扩散分析，其中６种可确定为富脯糖蛋白、淀粉酶、ＳＩｇＡ、酸性富脯蛋白、富酪蛋白及富组蛋白。实验结果表明唾液中的蛋白质选择性吸附到羟基磷灰石上，分离纯化的获得性膜蛋白为筛选促进致龋菌在牙面粘附的唾液蛋白受体提供了直接的研究材料。     

Binding of Actinomyces viscosus to collagen]

The ability of collagen adsorbed on apatitic surfaces to promote adhesion of A. viscosus was studied. Treatment of hydroxyapatite (HA) with human type I or type III collagen strongly promoted adhesion of A. viscosus LY7. TEM observations revealed that A. viscosus also attached to fibrils prepared from human type I collagen. The alpha(2) polypeptide derived from type I collagen exhibited moderate activity in promoting binding. Mutans of A. viscosus which possess type I fimbriae, but not type 2 fimbriae or no fimbriae, also bound to collagen-treated HA. This suggests that the adhesin responsible was associated with type I fimbriae, strains of A. israelii and A. odontolyticus also exhibited strong binding to collagen-treated HA. The avidity of Actinomyces species for collagen would seem to be partially responsible for the high proportions of these organisms found on root surfaces.     

粘性放线菌菌毛生物性能的研究：Ⅲ.粘性放线菌菌毛粘附与凝 …

目的 了解粘放菌两种菌毛在粘附牙面和与链球菌３４凝集中的生物活性,方法 采用粘放菌Ｉ型和ＩＩ型菌毛提取及两种抗菌毛抗体,通过它人对粘放菌粘附唾液包被的羟磷灰石（ＳＨＡ）的抑制实验来判断两种菌毛的粘附活性,通过粘放菌与血链球菌３４的凝集实验及两种抗菌毛抗体对凝集反应的抑制实验来确定两种菌毛的凝集活性。结果 （１）粘放菌Ｉ型和ＩＩ型菌毛提取物及两种抗菌毛抗体均能抑制粘放菌对ＳＨＡ的粘附,（２）只有ＩＩ     

Binding of Actinomyces viscosus to collagen: association with the type 1 fimbrial adhesin

Treatment of hydroxyapatite (HA) with human type I or type III collagen strongly promoted adhesion of Actinomyces viscosus LY7 cells. Treatment with human type V collagen was somewhat less effective while treatment with human type IV or rat type I collagen was significantly less effective. Electron microscopic observations revealed that A. viscosus cells also attached to fibrils prepared from human type I collagen. The alpha 1 (1) polypeptide chain derived from type I collagen was ineffective in promoting binding and the alpha 2 (1) polypeptide chain exhibited moderate activity. Heat- or urea-denatured type I collagens were also ineffective in promoting binding. Mutants of A. viscosus that possess type 1 fimbriae, but not type 2 fimbriae or no fimbriae, also bound to collagen-treated HA; this suggests that the adhesin responsible was associated with type 1 fimbriae. Strains of Actinomyces israelii and Actinomyces odontolyticus also exhibited strong binding to collagen-treated HA, while Actinomyces naeslundii ATCC 12104 did not. The avidity of Actinomyces species for collagen would seem to be at least partially responsible for the high proportions of these organisms found on cemental and root tooth surfaces.     

A study on the biological properties of fimbriae of Actinomyces viscosus. I. The preparation and determination of fimbriae of A. viscosus]

OBJECTIVE: To build up the methods to prepare and determine the fimbriae of A. viscosus. METHODS: Type 1 and 2 fimbriae were prepared from A. viscosus 5519 and 5951 respectively. Fimbriae were isolated from bacteria cells by mechanical shaking, obtained by collecting the supernatant after centrifugation, and purified by ammonium sulfate precipitations and gel filtration. RESULTS: Electron microscope showed that fimbriae existed in the preparations. The immunological reaction between the fimbriae preparations and specific antibodies against fimbriae attested that they were type 1 or type 2 fimbriae. CONCLUSION: The present study become a foundation for further study on the biological properties of fimbriae of A. viscosus.     

Analysis of the serum antibody responses to type 1 and type 2 fimbriae in mice immunized with Actinomyces viscosus T14V

The antibody responses of inbred mice immunized with Actinomyces viscosus T14V cells were analyzed using enzyme-linked immunoabsorbent assays (ELISAs) for measuring serum antibodies reactive with A. viscosus T14V cells and type 1 and type 2 fimbriae. In A/J mice immunized intraperitoneally on days 0 and 14, the serum antibody responses approached peak levels during d 19-35, and a dose of 10(8) cells/injection elicited optimal responses. Analysis of the responses of three genetically diverse strains of inbred mice indicated striking variations in the amounts of anti-type 1 (6.5-fold) and anti-type 2 (14.3-fold) antibodies elicited. The observed variations in the magnitude of the anti-fimbrial antibody responses are theoretically of sufficient magnitude to account for significant differences between mouse strains in their ability to inhibit adherence of A. viscosus T14V to saliva-coated hydroxyapatite and other bacteria. These studies provide a model with which the effects of variations in anti-fimbrial antibody responses on bacterial adherence may be analyzed.     

A study on biological properties of fimbriae of Actinomyces viscosus. II. Chemical characteristics of fimbriae of A. viscosus]

OBJECTIVE: To investigate the chemical characteristics of two purified fimbriae. METHODS: The specific antibodies against type I and type II fimbriae of A. viscosus were prepared and the purity of the two purified fimbriae had been identified by the specific antibodies. Then, chemical analysis and amino acid analysis of the two purified fimbriae were done. RESULTS: The two purified fimbriae were mainly composed of proteins and small amount of carbohydrate. The two types of fimbriae contained large amounts of aspartic acid, glutamic acid, alanine, lysine and glycine, and high percentage of nonpolar amino acid was analysed while basic amino acids were present in a very small amounts. CONCLUSION: These results may help to understand the relationship between the chemical characteristics of two types of fimbriae and their biological activity.     

粘性放线菌5519Ⅰ型菌毛的分离,鉴定

目的：对粘性放线菌５５１９Ⅰ型菌毛进行分离和部分特性研究。方法：采用超声粉碎方法从粘放菌５５１９细胞壁上分离Ⅰ型菌毛，然后经１６００００×ｇ２４ｈ高速离心，硫酸铵沉淀和葡聚糖凝胶Ｇ１００过柱后取得纯化菌毛，并经电镜和ＳＤＳ－ＰＡＧＥ电泳检查。结果：粘放菌５５１９Ⅰ型菌毛大部分分子量为５３５７３．４ｕ，少量为３７６９９．８ｕ。经电镜检查证实为Ⅰ型菌毛。结论：通过超声，结合高速离心，沉淀和葡聚糖凝胶Ｇ１００过程可得到纯化的粘放菌５５１９Ⅰ型菌毛     

Host modulation of adherence of oral bacteria: the effect of human neutrophil myeloperoxidase on the attachment of Actinomyces viscosus and Actinomyces naeslundii to saliva-coated hydroxyapatite

Host neutrophils, by virtue of their ability to secrete lysosomal enzymes and reduced oxygen metabolites, may play a fundamental role in altering the composition and pathogenic potential of dental plaque by modulating bacterial adherence. The adherence of Actinomyces viscosus and Actinomyces naeslundii to salivacoated surfaces is known to be saturable and mediated by proteinaceous fimbriae. In this study, we examined the abhesive effects of the human neutrophil enzyme, myeloperoxidase (MPO), on the adherence of A. viscosus T14V and A. naeslundii ATCC 12104 to saliva-coated hydroxyapatite (SHA) beads. Microorganisms were treated with MPO in the presence of hydrogen peroxide (H₂O₂) and physiologic concentrations of chloride (forming the MPO-H₂O₂-Cl- system) and incubated with SHA beads. Treatment of cells with the isolated MPO-H₂O₂-C1^- system reduced the percentage of cells that bound. This abhesive effect was inhibited by sodium azide, exogenous catalase, taurine, and potassium thiocyanate. The three major chromatographic forms of MPO (I, II, and III) were effective in blocking adherence of A. viscosus . We conclude that the MPO-H₂O₂-Cl^- system can interfere with the adherence of A. viscosus and A. naeslundii , and therefore may be important in modulating the microbiai colonization of oral surfaces.