Effects of extracellular slime produced by Staphylococcus epidermidis on oxidative responses of rabbit alveolar macrophages.
Effects of extracellular slime produced by Staphylococcus epidermidis on oxidative responses of rabbit alveolar macrophages.
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DOI：
10.3109/08941938909018263
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发表时间：
1989-01-01
期刊：
影响因子：
--
通讯作者：
Gristina, A G
中科院分区：
文献类型：
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作者：
Myrvik, Q N;Wagner, W;Gristina, A G
用4 M胍- hcl + 0.05 M醋酸钠和0.5% CHAPS提取表皮葡萄球菌RP 12菌株大量培养的细菌黏液，浓缩、透析，在DEAE sepacel柱上分离。用线性梯度NaCl洗脱I-2A、I-2B和I-4三个组分。分数I-2A和I-2B为阿辛蓝阳性，而分数I-4为阿辛蓝阴性，但电负性最强。将粗多糖部分和纯化后的三个部分分别与正常兔肺泡巨噬细胞（AM）孵养2.5或20 h，以确定它们对pma诱导的氧化破裂的影响。粗馏分（50-200微克/毫升）和I-2B（50-200微克/毫升）使AM在孵育2.5小时后，pma诱导的破裂增加了大约三倍。相比之下，与相同浓度的粗、I-2A或I-2B组分孵育20小时后，AM对pma诱导的爆发有30-40%的抑制作用。分数I-4没有明显的影响。还对这些组分进行了测试，以确定它们是否能引起bcg免疫AM的氧化爆发。即使bcg免疫AM产生的pma诱导的爆发是正常常驻AM的100倍，但没有任何组分（高达500微克/毫升）引起显著的氧化爆发。这些数据表明，表皮葡萄球菌的黏液可以在20小时的潜伏期内损害pma诱导的正常AM氧化爆发，这可以部分解释为什么宿主防御被产生黏液的表皮葡萄球菌破坏。
Bacterial slime produced in mass cultures of the RP 12 strain of Staphylococcus epidermidis was extracted with 4 M guanidine-HCl plus 0.05 M sodium acetate and 0.5% CHAPS, concentrated, dialyzed, and subjected to separation on DEAE sephacel columns. Three fractions, I-2A, I-2B, and I-4, were eluted with linear gradients of NaCl. Fractions I-2A and I-2B were alcian blue positive, whereas I-4 was alcian blue negative but the most electronegative fraction. The crude polysaccharide fraction and the three purified fractions were incubated individually for 2.5 or 20 h with normal rabbit alveolar macrophages (AM) to determine their effect on a subsequent PMA-induced oxidative burst. The crude fraction (50-200 micrograms/mL) and I-2B (50-200 micrograms/mL) primed the AM to give approximately a threefold increase in the PMA-induced burst after 2.5 h incubation. In contrast, a 20-h incubation resulted in a 30-40% inhibition of the PMA-induced burst with AM incubated with the same concentrations of the crude, I-2A, or I-2B fractions. Fraction I-4 had no detectable effect. The fractions also were tested to determine if they could elicit an oxidative burst in BCG-immune AM. None of the fractions (up to 500 micrograms/mL) elicited a significant oxidative burst even though BCG-immune AM yielded a PMA-induced burst 100 times that observed with normal resident AM. These data suggest that slime from S. epidermidis can impair the PMA-induced oxidative burst of normal AM during a 20-h incubation period and could explain in part why host defenses are compromised by slime-producing S. epidermidis.