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首页> 外文期刊>Infection and immunity >Mycoplasma hyorhinis molecules that induce tumor necrosis factor alpha secretion by human monocytes.
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Mycoplasma hyorhinis molecules that induce tumor necrosis factor alpha secretion by human monocytes.

机译：诱导人单核细胞分泌肿瘤坏死因子α的支原体分子。

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Mycoplasma hyorhinis has been shown to induce the secretion of tumor necrosis factor alpha (TNF-alpha) from monocytes. To identify the molecules responsible for this activity, we separated sonicated M. hyorhinis lysate material by centrifugation at 100,000 x g into soluble (S) and particulate (P) fractions. The fractions were assayed for TNF-alpha-inducing activity by the L929 bioassay. Both the soluble and particulate fractions were able to induce TNF-alpha in roughly equal amounts. The optimum dose for both fractions was 1 micrograms/ml. Proteinase K treatment of either fraction eliminated the activity, suggesting that a protein component is involved in induction. Phase partitioning into Triton X-114 aqueous (A) and detergent (D) phases showed that the soluble fraction was composed of 80% aqueous-phase proteins, while the particulate fraction was > 75% detergent-phase proteins. All four fractions (SA, SD, PA, and PD) were able to induce TNF-alpha release. Treatment with NaIO4 to remove carbohydrate reduced the inducing activity of the SA phase by 80%, whereas that of the other fractions was unaffected by this treatment. The M(r)S of the inducing activity were determined by the monocyte Western (immunoblot) technique. The SA phase activity was associated with a single periodate-sensitive peak of 69 to 75 kDa. The two detergent phases had similar profiles of inducing activity, containing four peaks of activity. These peaks corresponded to 48 to 52, 43 to 45, 39 to 40, and 31 to 32 kDa. The PA fraction also contained four peaks of activity, 69 to 75, 55 to 57, 48 to 52, and 39 to 40 kDa. Thus, both a protein and glycan moiety from M. hyorhinis are capable of inducing TNF-alpha release from human monocytes.
机译：业已证明，支原体可诱导单核细胞分泌肿瘤坏死因子α（TNF-α）。为了鉴定负责此活性的分子，我们通过以100,000 x g离心将超声处理的猪肺炎支原体裂解物分离为可溶性（S）和颗粒（P）馏分。通过L929生物测定法测定级分的TNF-α诱导活性。可溶部分和颗粒部分均能够诱导大致相等量的TNF-α。这两个部分的最佳剂量均为1微克/毫升。蛋白酶K处理任一部分都消除了该活性，表明蛋白质成分参与了诱导。相分离成Triton X-114水相（A）和去污剂（D），表明可溶级分由80％的水相蛋白组成，而颗粒级分则大于75％的去污相蛋白。所有四个部分（SA，SD，PA和PD）均能够诱导TNF-α释放。用NaIO 4处理以除去碳水化合物使SA相的诱导活性降低了80％，而其他部分的诱导活性不受该处理的影响。诱导活性的M（r）S通过单核细胞Western（免疫印迹）技术确定。 SA相活性与69至75kDa的单个高碘酸盐敏感峰相关。两个去污剂相具有相似的诱导活性曲线，包含四个活性峰。这些峰对应于48至52、43至45、39至40和31至32kDa。 PA级分还包含四个活性峰，分别为69至75、55至57、48至52和39至40 kDa。因此，来自透明支原体的蛋白和聚糖部分均能够诱导人单核细胞释放TNF-α。

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