Palmer Robert J, Gordon Sharon M, Cisar John O, Kolenbrander Paul E
National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892-4350, USA.
J Bacteriol. 2003 Jun;185(11):3400-9. doi: 10.1128/JB.185.11.3400-3409.2003.
Streptococci and actinomyces that initiate colonization of the tooth surface frequently coaggregate with each other as well as with other oral bacteria. These observations have led to the hypothesis that interbacterial adhesion influences spatiotemporal development of plaque. To assess the role of such interactions in oral biofilm formation in vivo, antibodies directed against bacterial surface components that mediate coaggregation interactions were used as direct immunofluorescent probes in conjunction with laser confocal microscopy to determine the distribution and spatial arrangement of bacteria within intact human plaque formed on retrievable enamel chips. In intrageneric coaggregation, streptococci such as Streptococcus gordonii DL1 recognize receptor polysaccharides (RPS) borne on other streptococci such as Streptococcus oralis 34. To define potentially interactive subsets of streptococci in the developing plaque, an antibody against RPS (anti-RPS) was used together with an antibody against S. gordonii DL1 (anti-DL1). These antibodies reacted primarily with single cells in 4-h-old plaque and with mixed-species microcolonies in 8-h-old plaque. Anti-RPS-reactive bacteria frequently formed microcolonies with anti-DL1-reactive bacteria and with other bacteria distinguished by general nucleic acid stains. In intergeneric coaggregation between streptococci and actinomyces, type 2 fimbriae of actinomyces recognize RPS on the streptococci. Cells reactive with antibody against type 2 fimbriae of Actinomyces naeslundii T14V (anti-type-2) were much less frequent than either subset of streptococci. However, bacteria reactive with anti-type-2 were seen in intimate association with anti-RPS-reactive cells. These results are the first direct demonstration of coaggregation-mediated interactions during initial plaque accumulation in vivo. Further, these results demonstrate the spatiotemporal development and prevalence of mixed-species communities in early dental plaque.
引发牙面定植的链球菌和放线菌常常彼此以及与其他口腔细菌共同聚集。这些观察结果引发了这样一种假说,即细菌间黏附会影响菌斑的时空发育。为了评估此类相互作用在体内口腔生物膜形成中的作用,针对介导共同聚集相互作用的细菌表面成分的抗体被用作直接免疫荧光探针,结合激光共聚焦显微镜,以确定在可回收釉质片上形成的完整人类菌斑内细菌的分布和空间排列。在属内共同聚集中,诸如戈登氏链球菌DL1这样的链球菌可识别其他链球菌(如口腔链球菌34)所携带的受体多糖(RPS)。为了确定正在形成的菌斑中链球菌的潜在相互作用亚群,一种抗RPS抗体(抗RPS)与一种抗戈登氏链球菌DL1抗体(抗DL1)一起使用。这些抗体主要与4小时龄菌斑中的单个细胞以及8小时龄菌斑中的混合菌种微菌落发生反应。抗RPS反应性细菌经常与抗DL1反应性细菌以及通过一般核酸染色区分的其他细菌形成微菌落。在链球菌与放线菌的属间共同聚集中,放线菌的2型菌毛可识别链球菌上的RPS。与内氏放线菌T14V的2型菌毛抗体(抗2型)发生反应的细胞比链球菌的任何一个亚群都要少得多。然而,与抗2型反应的细菌与抗RPS反应性细胞紧密相连。这些结果首次直接证明了体内初始菌斑积累过程中共同聚集介导的相互作用。此外,这些结果证明了早期牙菌斑中混合菌种群落的时空发育和流行情况。