Moritz A J, Cappelli D, Lantz M S, Holt S C, Ebersole J L
Department of Periodontics, The University of Texas Health Science Center, and Wilford Hall Medical Center, Lackland Air Force Base, San Antonio 78284, USA.
J Periodontol. 1998 Jun;69(6):686-97. doi: 10.1902/jop.1998.69.6.686.
Targeting bacterial virulence factors such as proteases for immunization may hold the key to limiting or preventing loss of attachment and alveolar bone in periodontal disease. This study examined the clinical, microbiological, and immununological responses following active immunization with a purified Porphyromonas gingivalis cysteine protease (porphypain-2) in the nonhuman primate (Nhp) Macaca fascicularis. One group of Nhp was immunized with porphypain-2 antigen while control Nhp received placebo injections. All Nhp were subjected to experimental gingivitis followed by ligature-induced periodontitis in a split-mouth design. An enzyme-linked immunosorbent assay demonstrated that immunization elicited a significantly elevated and specific IgG antibody response to both whole cell P. gingivalis (36-fold) and to porphypain-2 (194-fold). Checkerboard hybridization DNA analysis of subgingival plaque from ligated sextants demonstrated that 25% more Gram-negative anaerobic species became significantly elevated from baseline and at earlier timepoints in the control group than in the immununized group. Immunization with this protease did not suppress the emergence of P. gingivalis. Clinical indices showed few changes related to immunization. Alveolar bone density changes demonstrated a highly significant loss in ligated sextants compared to non-ligated sextants within the control group (P < 0.001), and a smaller but significant difference within the immunized group (P = 0.043). Comparison of ligated sextants only demonstrated more bone loss in the control group versus the immunized group (-13.07+/-9.51 versus -9.41+/-6.18; computer-assisted densitometric image analysis units +/- SD); the difference approached, but did not reach, significance. The results suggest that porphypain-2 may contribute to the pathogenic potential of the subgingival plaque microbiota in the Nhp model of ligature-induced periodontitis, and that active immunization with porphypain-2 appeared capable of altering this pathogenic response.
将诸如蛋白酶等细菌毒力因子作为免疫靶点，可能是限制或预防牙周病中附着丧失和牙槽骨丧失的关键。本研究在非人灵长类动物（Nhp）食蟹猕猴中，检测了用纯化的牙龈卟啉单胞菌半胱氨酸蛋白酶（卟啉蛋白酶-2）进行主动免疫后的临床、微生物学和免疫学反应。一组Nhp用卟啉蛋白酶-2抗原免疫，而对照Nhp接受安慰剂注射。所有Nhp均采用分口设计，先进行实验性牙龈炎，然后进行结扎诱导的牙周炎。酶联免疫吸附测定表明，免疫引发了针对牙龈卟啉单胞菌全细胞（36倍）和卟啉蛋白酶-2（194倍）的显著升高的特异性IgG抗体反应。对结扎牙位的龈下菌斑进行棋盘杂交DNA分析表明，与免疫组相比，对照组中革兰氏阴性厌氧菌种类从基线水平显著升高且在更早时间点升高的比例多25%。用这种蛋白酶免疫并未抑制牙龈卟啉单胞菌的出现。临床指标显示与免疫相关的变化很少。牙槽骨密度变化表明，与对照组中未结扎的牙位相比，结扎牙位有极显著的骨量丧失（P < 0.001），而免疫组内差异较小但显著（P = 0.043）。仅比较结扎牙位发现，对照组的骨丧失比免疫组更多（-13.07±9.51对-9.41±6.18；计算机辅助密度测量图像分析单位±标准差）；差异接近但未达到显著水平。结果表明，在结扎诱导的牙周炎的Nhp模型中，卟啉蛋白酶-2可能有助于龈下菌斑微生物群的致病潜力，并且用卟啉蛋白酶-2进行主动免疫似乎能够改变这种致病反应。
