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首页> 外文期刊>Infection and immunity >Borrelia burgdorferi Induces the Production and Release of Proinflammatory Cytokines in Canine Synovial Explant Cultures
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Borrelia burgdorferi Induces the Production and Release of Proinflammatory Cytokines in Canine Synovial Explant Cultures

机译：伯氏疏螺旋体诱导犬滑膜外植体培养物中促炎细胞因子的产生和释放

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Canine synovial membrane explants were exposed to high- or low-passage Borrelia burgdorferi for 3, 6, 12, and 24 h. Spirochetes received no treatment, were UV light irradiated for 16 h, or were sonicated prior to addition to synovial explant cultures. In explant tissues, mRNA levels for the proinflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-1α (IL-1α), IL-1β, and IL-8 were surveyed semiquantitatively by reverse transcription-PCR. Culture supernatants were examined for numbers of total and motile (i.e., viable) spirochetes, TNF-like and IL-1-like activities, polymorphonuclear neutrophil (PMN) chemotaxis-inducing activities, and IL-8. During exposure to synovial explant tissues, the total number of spirochetes in the supernatants decreased gradually by ～30%, and the viability also declined. mRNAs for TNF-α, IL-1α, IL-1β, and IL-8 were up-regulated in synovial explant tissues within 3 h after infection with untreated or UV light-irradiated B. burgdorferi, and mRNA levels corresponded to the results obtained with bioassays. During 24 h of coincubation, cultures challenged with untreated or UV light-irradiated spirochetes produced similar levels of TNF-like and IL-1-like activities. In contrast, explant tissues exposed to untreatedB. burgdorferi generated significantly higher levels of chemotactic factors after 24 h of incubation than did explant tissues exposed to UV light-treated spirochetes. In identical samples, a specific signal for IL-8 was identified by Western blot analysis. High- and low-passage borreliae did not differ in their abilities to induce proinflammatory cytokines. No difference in cytokine induction between untreated and sonicated high-passage spirochetes was observed, suggesting that fractions of the organism can trigger the production and release of inflammatory mediators. The titration of spirochetes revealed a dose-independent cytokine response, where 103 to 107 B. burgdorferi organisms induced similar TNF-like activities but only 107 spirochetes induced measurable IL-1-like activities. The release of chemotactic factors was dose dependent and was initiated when tissues were infected with at least 105 organisms. We conclude that intact B. burgdorferi or fractions of the bacterium can induce the local up-regulation of TNF-α, IL-1α, and IL-1β in the synovium but that the interaction of viable spirochetes with synovial cells leads to the release of IL-8, which probably is a prime initiator of PMN migration during acute Lyme arthritis.
机译：将犬滑膜外植体暴露于高通道或低通道 Borrelia burgdorferi 3、6、12和24小时。螺旋体未接受处理，未用紫外线照射16 h，或在加入滑膜外植体培养物之前进行了超声处理。在外植体组织中，通过逆转录PCR半定量检测促炎细胞因子肿瘤坏死因子α（TNF-α），白细胞介素1α（IL-1α），IL-1β和IL-8的mRNA水平。检查培养物上清液的总和运动的（即活的）螺旋体的数目，TNF样和IL-1样活性，多形核中性粒细胞（PMN）趋化性诱导活性和IL-8。暴露于滑膜外植体组织期间，上清液中的螺旋体总数逐渐减少了约30％，活力也下降了。在未经治疗或经紫外线照射的 B感染后3小时内，滑膜外植体组织中TNF-α，IL-1α，IL-1β和IL-8的mRNA上调。 burgdorferi 和mRNA水平与生物测定的结果相对应。在共孵育的24小时内，用未经处理或紫外线照射的螺旋体攻击的培养物产生相似水平的TNF-like和IL-1-like活性。相反，外植体组织暴露于未经处理的B。孵育24小时后，burgdorferi 产生的趋化因子水平明显高于暴露于紫外线照射的螺旋体的外植体组织。在相同的样品中，通过蛋白质印迹分析鉴定了IL-8的特异性信号。高通道和低通道疏螺旋体诱导促炎细胞因子的能力没有差异。在未处理的和超声处理的高通道螺旋体之间未观察到细胞因子诱导的差异，这表明该生物体的各个部分可以触发炎症介质的产生和释放。螺旋体的滴定显示出剂量依赖性的细胞因子反应，其中10 3 至10 7 B。 burgdorferi 生物诱导了类似的TNF-like活性，但只有10 7 螺旋体诱导了可测量的IL-1活性。趋化因子的释放是剂量依赖性的，并且是在组织感染至少10 5 生物后才开始释放的。我们得出结论，完整的 B。 burgdorferi 或部分细菌可诱导滑膜中TNF-α，IL-1α和IL-1β的局部上调，但活螺旋体与滑膜细胞的相互作用导致IL-的释放图8可能是急性莱姆关节炎期间PMN迁移的主要起因。

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