Differential induction of inflammatory cytokines by dendritic cells treated with novel TLR-agonist and cytokine based cocktails: targeting dendritic cells in autoimmunity.
Differential induction of inflammatory cytokines by dendritic cells treated with novel TLR-agonist and cytokine based cocktails: targeting dendritic cells in autoimmunity.
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DOI：
10.1186/1476-9255-7-37
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发表时间：
2010-07-27
期刊：
影响因子：
--
通讯作者：
Gad M
中科院分区：
文献类型：
--
作者：
Jensen SS;Gad M
树突状细胞（DC）是免疫系统的主要看门人，连接先天免疫系统和适应性免疫系统。无论是自身免疫性疾病还是过敏性疾病，dc都能够在炎症部位成熟为炎性dc，从而维持炎症部位适应性免疫系统的持续激活。树突状细胞的这种功能使它们成为炎症性疾病治疗干预的有吸引力的靶细胞。我们设计了一个基于DC的筛选模型，通过该模型可以评估候选药物抑制DC成熟为炎症和疾病促进表型的能力。利用IL-4和GM-CSF将人单核细胞来源的dc分化为未成熟的dc （imdc）。用tlr激动剂和促炎细胞因子的各种组合处理imdc，以鉴定能够将imdc成熟为炎性dc的鸡尾酒。利用ELISA和细胞因子阵列检测分泌的细胞因子和趋化因子，评估鸡尾酒对DC成熟的影响。使用FACS分析来评估成熟标志物的表达，并使用naïve同种异体t细胞进行功能研究，以检测th1促进DC表型。9种鸡尾酒能诱导imdc分泌促th1的细胞因子IL-12p70和tnf - α， 3种鸡尾酒能诱导促th17的细胞因子IL-23。使用细胞因子阵列进一步表征鸡尾酒，显示诱导炎症相关细胞因子和趋化因子，如CXCL10， CCL2, CCL4, CCL8, CCL15， CCL20和IL-8，其中一些存在于一系列自身免疫性疾病中。经TLR激动剂poly I:C和肽聚糖处理的树突状细胞可分泌前列腺素E2，但LPS未发现。除了TLR7/8激动剂R848外，鸡尾酒能够诱导DC成熟标记物如HLA-DR、CD40、CD80、CD83和CD86。同种异体CD4+ T细胞与经鸡尾酒处理的树突状细胞共培养的功能终点表明，五种鸡尾酒可以诱导典型的th1表型，并能够分泌大量的标志性细胞因子IFNγ。该模型使用地塞米松和两种cox -抑制剂进行验证，这两种抑制剂能够抑制鸡尾酒驱动的促炎性DC成熟。通过评估抑制imdc向具有特定th1促进表型的炎性dc的激活和分化的能力，鉴定新的th1促进鸡尾酒可以筛选抗炎候选药物。因此，该模型提供了一种筛选工具，可以识别对免疫反应的自然调节剂树突状细胞的潜在抗炎作用。
Dendritic cells (DC) are main gate-keepers of the immune system, bridging the innate and adaptive immune system. DCs are able to mature into inflammatory DCs at sites of inflammation in both autoimmune and allergic disease, thereby sustaining a continuous activation of the adaptive immune system at sites of inflammation. This function of DCs makes them attractive target cells for therapeutic intervention in inflammatory diseases. We have designed a DC-based screening model by which drug candidates can be evaluated for their ability to suppress DC maturation into an inflammatory and disease promoting phenotype. Human monocyte derived DCs were differentiated using IL-4 and GM-CSF to immature DCs (imDCs). The imDCs were treated with various combinations of TLR-agonists and pro-inflammatory cytokines to identify cocktails with ability to mature imDCs into inflammatory DCs. The effect of the cocktails on DC maturation was evaluated using ELISA and cytokine arrays to measure secreted cytokines and chemokines. FACS analysis was used to assess expression of maturation markers, and functional studies were carried out using naïve allogeneic T-cells to assay for a Th1-promoting DC phenotype. Nine cocktails were designed with potent ability to induce secretion of the Th1-promoting cytokines IL-12p70 and TNFα from imDCs, and three were able to induce the Th17-promoting cytokine IL-23. The cocktails were further characterized using cytokine arrays, showing induction of inflammation related cytokines and chemokines like CXCL10, CCL2, CCL4, CCL8, CCL15, CCL20 and IL-8, of which some are present in a range of autoimmune pathologies. Prostaglandin E2 secretion was identified from DCs treated with TLR agonists poly I:C and peptidoglycan, but not LPS. The cocktails were able to induce DC maturation markers like HLA-DR, CD40, CD80, CD83 and CD86, except the TLR7/8 agonist R848. Functional end-points made by co-culture of allogeneic CD4+ T cells with the cocktail treated DCs, showed that five cocktails in particular could induce a classical Th1-phenotype with ability to secrete high amounts of the hall-mark cytokine IFNγ. The model was validated using dexamethasone and two COX-inhibitors, which were able to suppress the cocktail driven pro-inflammatory DC maturation. The identification of novel Th1-promoting cocktails allows screening of anti-inflammatory drug candidates by assessing the ability to suppress the activation and differentiation of imDCs into inflammatory DCs with a specific Th1-promoting phenotype. The model thus provides a screening tool, which can identify potential anti-inflammatory effects on the natural regulator of the immune response, the dendritic cell.
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影响因子：
4.9
作者：
Hanauer, SB
通讯作者：
Hanauer, SB
影响因子：
30.5
作者：
Langenkamp, A;Messi, M;Sallusto, F
通讯作者：
Sallusto, F
影响因子：
4.4
作者：
Pradhan, Sanjay;Genebriera, Joseph;Timares, Laura
通讯作者：
Timares, Laura
DOI：
10.1038/nri2335
发表时间：
2008-06
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
通讯作者：
--
影响因子：
100.3
作者：
Bachmann, MF;Kopf, M;Marsland, BJ
通讯作者：
Marsland, BJ