Takei Masao, Nakagawa Hideyuki
Division of Cellular Allergology, Research Center Borstel, Parkallee 22, D-23845 Borstel, Germany.
Toxicol Appl Pharmacol. 2006 May 15;213(1):27-36. doi: 10.1016/j.taap.2005.08.004. Epub 2005 Sep 28.
The sea urchin Toxopneustes pileolus belonging to the family Toxopneustidae, they have well-developed globiferous pedicellariae with pharmacologically active substances. We have purified a novel sea urchin lectin-1 (SUL-1) from the large globiferous pedicellariae of T. pileolus. Dendritic cells (DC) are professional APC and play a pivotal role in controlling immune responses. This study investigated whether SUL-1 can drive DC maturation from human immature monocyte-derived DC in vitro. Human monocytes were cultured with GM-CSF and IL-4 for 6 days followed by another 1 day in the presence of SUL-1 or LPS. DC harvested on day 7 were examined using functional assays. The expression levels of CD1a, CD80, CD83, CD86 and HLA-DR as expressed by mean fluorescence intensity (MFI) on DC differentiated from immature DC after culture with 1.0 microg/ml of SUL-1 for 1 day were enhanced and decreased endocytic activity. SUL-1-treated DC also displayed enhanced T cell stimulatory capacity in an MLR, as measured by T cell proliferation. Cell surface expression of CD80, CD83 and CD86 on SUL-1-treated DC was inhibited by anti-DC-SIGN mAb, while anti-DC-SIGN mAb had no influence on allogeneic T cell proliferation by SUL-1-treated DC. DC differentiated with SUL-1 induced the differentiation of naïve T cell towards a helper T cell type 1 (Th1) response at DC/T (1:5) cells ratio depending on IL-12 secretion. In CTL assay, the production of IFN-gamma and 51Cr release on SUL-1-treated DC were more augmented than of immature DC or LPS-treated DC. SUL-1-treated DC expressed CCR7 and had a high migration to MIP-3beta. Intracellular Ca2+ mobilization in SUL-1-treated DC was also induced by MIP-3beta. These results suggest that SUL-1 bindings to DC-SIGN on surface of immature DC may lead to differentiate DC from immature DC. Moreover, it suggests that SUL-1 may be used on DC-based vaccines for cancer immunotherapy.
隶属于毒棘海胆科的毒棘海胆，其拥有发育良好的含毒腺叉棘，且含有药理活性物质。我们从毒棘海胆的大型含毒腺叉棘中纯化出了一种新型海胆凝集素-1（SUL-1）。树突状细胞（DC）是专职抗原呈递细胞，在控制免疫反应中起关键作用。本研究调查了SUL-1是否能在体外驱动人未成熟单核细胞来源的DC成熟。将人单核细胞与GM-CSF和IL-4培养6天，然后在SUL-1或LPS存在的情况下再培养1天。使用功能测定法检测第7天收获的DC。在用1.0微克/毫升SUL-1培养1天后，从未成熟DC分化而来的DC上，以平均荧光强度（MFI）表示的CD1a、CD80、CD83、CD86和HLA-DR的表达水平升高，且内吞活性降低。通过T细胞增殖测量，SUL-1处理的DC在混合淋巴细胞反应（MLR）中也表现出增强的T细胞刺激能力。抗DC-SIGN单克隆抗体抑制了SUL-1处理的DC上CD80、CD83和CD86的细胞表面表达，而抗DC-SIGN单克隆抗体对SUL-1处理的DC的同种异体T细胞增殖没有影响。用SUL-1分化的DC在DC/T（1:5）细胞比例下，根据IL-12分泌情况，诱导幼稚T细胞向辅助性T细胞1型（Th1）反应分化。在细胞毒性T淋巴细胞（CTL）测定中，SUL-1处理的DC上IFN-γ 的产生和51Cr释放比未成熟DC或LPS处理的DC增加得更多。SUL-1处理的DC表达CCR7，并且对MIP-3β 具有高迁移性。MIP-3β 也诱导了SUL-1处理的DC中的细胞内Ca2+ 动员。这些结果表明，SUL-1与未成熟DC表面的DC-SIGN结合可能导致DC从未成熟DC分化而来。此外，这表明SUL-1可用于基于DC的癌症免疫治疗疫苗。
