[Chemical structure and immunobiological activities of peptidoglycan isolated from Capnocytophaga species].

T Hanagata
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Abstract

The chemical structure and immunobiological activities of the cell wall peptidoglycan isolated from Capnocytophaga species was investigated. Peptidoglycan was isolated from Capnocytophaga species strain SE2-2 by boiling in 4% sodium dodecyl sulfate and by digestion with pronase, trypsin and alpha-amylase. Analysis of amino acids and amino sugars of the peptidoglycan revealed that glucosamine, muramic acid, D-glutamic acid, alanine, and diaminopimelic acid (A2pm) were the principal components. Serine and glycine were not found. Dinitrophenylation method revealed that about half of A2pm residue had a free amino group, and analysis by hydrazinolysis showed that a small part of alanine and A2pm located at the C-terminal. The above results indicate that one of the amino groups of A2pm residue at one strand of the stem peptide subunit cross-linked to the carboxyl group of alanine of the neighboring strand. It was thus revealed that the peptidoglycan of Capnocytophaga species belonged to the Al gamma type of the classification by Schleifer and Kandler. Peptidoglycan isolated from Capnocytophaga species strain SE2-2 was found to be definitely adjuvant-active in induction of delayed type hypersensitivity against ovalbumin when administered to guinea pigs as water-in-oil emulsion and in stimulation of increase serum antibody levels. Regarding mitogenicity on splenocytes of BALB/c and BALB/c nu/nu mice, peptidoglycan from Capnocytophaga species was markedly enhanced the uptake [3H] thymidine in dose of 10 micrograms/10(5) cells, however thymocytes were not reactive. Stimulation effects on peritoneal macrophages from a guinea pig to incorporation of 14C-glucosamin was exhibited by addition of 100 micrograms of this peptidoglycan. These findings indicate that peptidoglycan of Capnocytophaga species might eventually be responsible for destruction of periodontal tissue by host mediated activities.

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[从碳吞噬菌中分离的肽聚糖的化学结构和免疫生物学活性]
研究了从碳吞噬菌中分离的细胞壁肽聚糖的化学结构和免疫生物学活性。用4%十二烷基硫酸钠煮沸,经蛋白酶、胰蛋白酶和α -淀粉酶消化,从Capnocytophaga菌株SE2-2中分离得到肽聚糖。肽聚糖的氨基酸和氨基糖分析表明,葡萄糖胺、氨基乙酸、d -谷氨酸、丙氨酸和二氨基戊酸(A2pm)是主要成分。没有发现丝氨酸和甘氨酸。二硝基苯基化分析表明,A2pm残基中约有一半有游离氨基,肼解分析表明,有一小部分丙氨酸和A2pm位于c端。上述结果表明,A2pm残基在茎肽亚基的一条链上的一个氨基与相邻链的丙氨酸羧基交联。结果表明,碳吞噬菌属的肽聚糖属Schleifer和Kandler分类中的Al γ型。从Capnocytophaga菌株SE2-2中分离的肽聚糖被发现在诱导对卵清蛋白的延迟型超敏反应中具有明确的佐剂活性,当作为油包水乳剂给予豚鼠时,可以刺激血清抗体水平的增加。在BALB/c和BALB/c nu/nu小鼠脾细胞的有丝分裂性方面,来自Capnocytophaga的肽聚糖在10微克/10(5)个细胞的剂量下显著促进了[3H]胸腺嘧啶的摄取,但胸腺细胞没有反应。在豚鼠腹膜巨噬细胞中加入100微克的14c -氨基葡萄糖肽聚糖,显示出其对腹腔巨噬细胞的刺激作用。这些发现表明,噬碳菌的肽聚糖可能最终通过宿主介导的活动破坏牙周组织。
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