A rapid fluorescence-based assay for classification of iNKT cell activating glycolipids.
A rapid fluorescence-based assay for classification of iNKT cell activating glycolipids.
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DOI：
10.1021/ja200070u
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发表时间：
2011-04-13
影响因子：
15
通讯作者：
Porcelli, Steven A.
中科院分区：
文献类型：
--
作者：
Arora, Pooja;Venkataswamy, Manjunatha M.;Baena, Andres;Bricard, Gabriel;Li, Qian;Veerapen, Natacha;Ndonye, Rachel;Park, Jeong Ju;Lee, Ji Hyung;Seo, Kyung-Chang;Howell, Amy R.;Chang, Young-Tae;Illarionov, Petr A.;Besra, Gurdyal S.;Chung, Sung-Kee;Porcelli, Steven A.
激活不变自然杀伤 T 细胞（iNKT 细胞）的 α-半乳糖神经酰胺 (αGC) 的结构变体正在被开发为潜在的免疫调节剂，用于各种应用。鉴定这些糖脂的特定形式是当前努力的核心，这些糖脂的反应偏向于产生促炎细胞因子和抗炎细胞因子，但由于缺乏可靠预测这些化合物体内生物活性的体外筛选测定，这一目标受到了阻碍。在这里，我们描述了一种基于荧光的测定法来识别功能不同的 αGC 类似物。我们的测定基于最近的研究结果，表明定位于质膜去污剂抗性微结构域（脂筏）的 CD1d 分子呈递糖脂抗原与干扰素 γ 分泌和 Th1 偏向细胞因子反应的诱导相关。使用一种测量装载 αGC 的 CD1d 分子脂筏驻留的测定方法，我们筛选了约 200 种合成 αGC 类似物的文库，并鉴定了 19 种具有潜在 Th1 偏向活性的激动剂。对这些新型候选 Th1 型激动剂的一部分在小鼠体内的分析证实了它们诱导与 Th1 型偏倚一致的全身细胞因子反应的能力。这些结果证明了这种新颖的体外测定对于评估糖脂激动剂的体内功能的预测价值，并为用于 iNKT 细胞活化糖脂的鉴定和功能分类的相对简单的高通量测定提供了基础。
Structural variants of α-galactosylceramide (αGC) that activate invariant natural killer T cells (iNKT cells) are being developed as potential immunomodulatory agents for a variety of applications. Identification of specific forms of these glycolipids that bias responses to favor production of proinflammatory vs anti-inflammatory cytokines is central to current efforts, but this goal has been hampered by the lack of in vitro screening assays that reliably predict the in vivo biological activity of these compounds. Here we describe a fluorescence-based assay to identify functionally distinct αGC analogues. Our assay is based on recent findings showing that presentation of glycolipid antigens by CD1d molecules localized to plasma membrane detergent-resistant microdomains (lipid rafts) is correlated with induction of interferon-γ secretion and Th1-biased cytokine responses. Using an assay that measures lipid raft residency of CD1d molecules loaded with αGC, we screened a library of ∼200 synthetic αGC analogues and identified 19 agonists with potential Th1-biasing activity. Analysis of a subset of these novel candidate Th1 type agonists in vivo in mice confirmed their ability to induce systemic cytokine responses consistent with a Th1 type bias. These results demonstrate the predictive value of this novel in vitro assay for assessing the in vivo functionality of glycolipid agonists and provide the basis for a relatively simple high-throughput assay for identification and functional classification of iNKT cell activating glycolipids.
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影响因子：
7.8
作者：
Venkataswamy MM;Porcelli SA
通讯作者：
Porcelli SA
影响因子：
4.4
作者：
Forestier, Claire;Takaki, Toshiyuki;Porcelli, Steven A.
通讯作者：
Porcelli, Steven A.
影响因子：
0.6
作者：
Mori, Kenji;Tashiro, Takuya
通讯作者：
Tashiro, Takuya
DOI：
10.4049/jimmunol.0900858
发表时间：
2009-08-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Venkataswamy MM;Baena A;Goldberg MF;Bricard G;Im JS;Chan J;Reddington F;Besra GS;Jacobs WR Jr;Porcelli SA
通讯作者：
Porcelli SA
影响因子：
32.4
作者：
Im, Jin S.;Arora, Pooja;Bricard, Gabriel;Molano, Alberto;Venkataswamy, Manjunatha M.;Baine, Ian;Jerud, Elliot S.;Goldberg, Michael F.;Baena, Andres;Yu, Karl O. A.;Ndonye, Rachel M.;Howell, Amy R.;Yuan, Weiming;Cresswell, Peter;Chang, Young-tae;Illarionov, Petr A.;Besra, Gurdyal S.;Porcelli, Steven A.
通讯作者：
Porcelli, Steven A.