ORAL PASSIVE-IMMUNIZATION AGAINST DENTAL-CARIES IN RATS BY USE OF HEN EGG-YOLK ANTIBODIES SPECIFIC FOR CELL-ASSOCIATED GLUCOSYLTRANSFERASE OF STREPTOCOCCUS-MUTANS
ORAL PASSIVE-IMMUNIZATION AGAINST DENTAL-CARIES IN RATS BY USE OF HEN EGG-YOLK ANTIBODIES SPECIFIC FOR CELL-ASSOCIATED GLUCOSYLTRANSFERASE OF STREPTOCOCCUS-MUTANS
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DOI：
10.1128/iai.59.11.4161-4167.1991
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发表时间：
1991-11-01
影响因子：
3.1
通讯作者：
OOSHIMA, T
中科院分区：
文献类型：
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作者：
HAMADA, S;HORIKOSHI, T;OOSHIMA, T
用抗血清型c变异链球菌全细胞、无细胞（CF）葡萄糖基转移酶（GTase）或细胞相关（CA）GTase的多克隆卵黄免疫球蛋白G抗体（yIgG）对变异链球菌毒力相关因子的体外抑制作用进行了研究。变形链球菌和保护S.抗龋齿的发展。 用福尔马林处理的全细胞、纯化的CF-GT酶或CA-GT酶连同弗氏完全佐剂免疫母鸡（18周龄）。 此外，在一些体外实验中使用针对表面蛋白抗原的yIgG以与其他抗体进行比较。 通过硫酸铵沉淀，然后通过DEAE-Sephacel柱层析或用乙醇分级沉淀来纯化yIgG。 发现纯化的yIgG是220-kDa的蛋白质，其在加入2-巯基乙醇后解离成重链和轻链。 yIgG对全细胞和表面蛋白抗原产生大量聚集。而CF-和CA-GT酶的yIgG特异性抑制相应GT酶的酶活性。 抗CA-GT酶和全细胞的yIgG明显抑制S.变形细胞贴在玻璃表面。 无特异性病原体的Sprague-Dawley大鼠被严重和反复感染S。变形链球菌和喂食2000号饮食的大鼠出现严重的龋齿，而喂食含有大于或等于0.1%的抗CA-GT酶γ IgG的2000号饮食的大鼠显示出牙菌斑积累和龋齿发展的统计学显著减少。 向CF-GTase和全细胞施用yIgG未能预防龋齿。 这些结果清楚地表明，抗S.变形菌CA-GT酶特异性抑制该生物体的毒力因子，即，不溶性葡聚糖合成CA-GT酶，导致龋齿的发展显着减少。
The effect of polyclonal egg yolk immunoglobulin G antibodies (yIgG) raised against whole cells, cell-free (CF) glucosyltransferase (GTase), or cell-associated (CA) GTase of serotype c Streptococcus mutans was examined in terms of in vitro inhibition of virulence-related factors of S. mutans and protection of S. mutans-infected rats against the development of dental caries. Hens (18 weeks old) were immunized with formalin-treated whole cells, purified CF-GTase, or CA-GTase together with Freund's complete adjuvant. In addition, yIgG to surface protein antigen was used in some in vitro experiments for comparison with other antibodies. yIgG was purified by ammonium sulfate precipitation followed by DEAE-Sephacel column chromatography or fractional precipitation with ethanol. Purified yIgG was found to be a 220-kDa protein, which was dissociated into heavy and light chains upon addition of 2-mercaptoethanol. yIgG to whole cells and surface protein antigen gave a heavy aggregation of S. mutans organisms, while yIgG to CF- and CA-GTase specifically inhibited the enzymatic activity of the respective GTase. yIgG to CA-GTase and whole cells was found to clearly suppress the adherence of S. mutans cells to a glass surface. Specific-pathogen-free Sprague-Dawley rats that had been infected heavily and repeatedly with S. mutans and fed diet no. 2000 developed severe dental caries, while rats fed diet 2000 containing greater-than-or-equal-to 0.1% yIgG to CA-GTase showed a statistically significant reduction in dental plaque accumulation and caries development. Administration of yIgG to CF-GTase and whole cells failed to protect against caries. These results clearly suggest that yIgG to S. mutans CA-GTase specifically inhibited a virulence factor of this organism, i.e., insoluble glucan-synthesizing CA-GTase, resulting in a significant reduction in the development of dental caries.