The immunoregulatory role of alpha enolase in dendritic cell function during Chlamydia infection.
The immunoregulatory role of alpha enolase in dendritic cell function during Chlamydia infection.
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DOI：
10.1186/s12865-017-0212-1
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发表时间：
2017-05-19
期刊：
影响因子：
3
通讯作者：
He Q
中科院分区：
文献类型：
--
作者：
Ryans K;Omosun Y;McKeithen DN;Simoneaux T;Mills CC;Bowen N;Eko FO;Black CM;Igietseme JU;He Q
我们以前曾报道，白细胞介素-10（IL-10）缺陷的树突状细胞（DC）是有效的抗原提呈细胞，诱导提高保护性免疫对衣原体。为了进一步研究IL-10缺陷DC的上级免疫刺激特性的分子和生化机制，我们对衣原体脉冲的野生型（WT）和IL-10−/− DC的蛋白质谱进行了蛋白质组学分析，以鉴定具有免疫调节特性的差异表达蛋白。结果显示，与WT DC相比，α烯醇化酶（ENO 1），一种参与糖酵解最后一步的代谢酶，在衣原体脉冲的IL-10−/− DC中显著上调。我们通过使用慢病毒siRNA技术产生ENO 1敲低的DC，进一步研究了ENO 1在DC功能中的免疫调节作用。我们分析了用衣原体脉冲后ENO 1敲低对DC功能的影响。丙酮酸测定、透射电镜、流式细胞术、共聚焦显微镜、细胞因子、T细胞活化和过继转移测定也用于研究DC功能。结果显示，与衣原体冲击的WT DC相比，ENO 1敲低的DC具有受损的成熟和活化，细胞内丙酮酸浓度显著降低。衣原体致敏的ENO 1敲低DC的连续转移在体内外均缺乏免疫原性，尤其是诱导针对生殖道衣原体感染的保护性免疫的能力。与衣原体脉冲的WT DC相比，衣原体脉冲的ENO 1敲低DC的线粒体形态的显著重塑与调节线粒体通透性的外膜转位酶（TOM）20和腺嘌呤核苷酸转位酶（ANT）1/2/3/4的失调相关。结果表明，增强的糖酵解是DC有效加工和呈递抗原以诱导强大的免疫应答所必需的。ENO 1的上调有助于IL-10缺陷型DC的上级免疫刺激功能。我们的研究表明，ENO 1缺乏导致丙酮酸的产生减少，然后导致线粒体稳态功能障碍，可能影响DC的存活，成熟和抗原呈递特性。因此，调节ENO 1提供了一种潜在有效的策略来增强DC功能并促进针对感染性和非感染性疾病的免疫力。
We have previously reported that interleukin-10 (IL-10) deficient dendritic cells (DCs) are potent antigen presenting cells that induced elevated protective immunity against Chlamydia. To further investigate the molecular and biochemical mechanism underlying the superior immunostimulatory property of IL-10 deficient DCs we performed proteomic analysis on protein profiles from Chlamydia-pulsed wild-type (WT) and IL-10−/− DCs to identify differentially expressed proteins with immunomodulatory properties. The results showed that alpha enolase (ENO1), a metabolic enzyme involved in the last step of glycolysis was significantly upregulated in Chlamydia-pulsed IL-10−/− DCs compared to WT DCs. We further studied the immunoregulatory role of ENO1 in DC function by generating ENO1 knockdown DCs, using lentiviral siRNA technology. We analyzed the effect of the ENO1 knockdown on DC functions after pulsing with Chlamydia. Pyruvate assay, transmission electron microscopy, flow cytometry, confocal microscopy, cytokine, T-cell activation and adoptive transfer assays were also used to study DC function. The results showed that ENO1 knockdown DCs had impaired maturation and activation, with significant decrease in intracellular pyruvate concentration as compared with the Chlamydia-pulsed WT DCs. Adoptive transfer of Chlamydia-pulsed ENO1 knockdown DCs were poorly immunogenic in vitro and in vivo, especially the ability to induce protective immunity against genital chlamydia infection. The marked remodeling of the mitochondrial morphology of Chlamydia-pulsed ENO1 knockdown DCs compared to the Chlamydia-pulsed WT DCs was associated with the dysregulation of translocase of the outer membrane (TOM) 20 and adenine nucleotide translocator (ANT) 1/2/3/4 that regulate mitochondrial permeability. The results suggest that an enhanced glycolysis is required for efficient antigen processing and presentation by DCs to induce a robust immune response. The upregulation of ENO1 contributes to the superior immunostimulatory function of IL-10 deficient DCs. Our studies indicated that ENO1 deficiency causes the reduced production of pyruvate, which then contributes to a dysfunction in mitochondrial homeostasis that may affect DC survival, maturation and antigen presenting properties. Modulation of ENO1 thus provides a potentially effective strategy to boost DC function and promote immunity against infectious and non-infectious diseases.