Vray Bernard, Camby Isabelle, Vercruysse Vincent, Mijatovic Tatjana, Bovin Nicolai V, Ricciardi-Castagnoli Paola, Kaltner Herbert, Salmon Isabelle, Gabius Hans-Joachim, Kiss Robert
Laboratoire d'Immunologie Expérimentale, Faculté de Médecine, 808 Route de Lennik, 1070 Brussels, Belgium.
Glycobiology. 2004 Jul;14(7):647-57. doi: 10.1093/glycob/cwh068. Epub 2004 Mar 24.
The impact of a pathogen-induced inflammatory response on dendritic cells (DCs) and on their expression of galectin-3 (Gal-3) was studied on splenic DCs (sDCs) from Trypanosoma cruzi-infected mice. We determined the lectin expression and also presentation of ligands using the labeled galectin as probe. By reverse transcriptase polymerase chain reaction, western blot analysis, quantitative glycocytochemistry, and computer-assisted quantitative microscopy, we demonstrate that, in sDCs from infected mice, expression of Gal-3 and Gal-3-specific ligands were markedly up-regulated and adhesiveness was increased with Gal-3-coated substratum. Gal-3 expression was also enhanced in T. cruzi-infected D2SC-1 cells. To assess influence on migration, we had to work exclusively with D2SC-1 cells because sDCs rapidly lost their capacity to adhere to substratum. Migration of infected- and TCM-treated D2SC-1 cells were reduced when substratum was coated with Gal-3. Expression of Gal-3 by D2SC-1 was reduced when they were incubated with anti-Gal-3 antisense oligonucleotide without effect on cell invasion by the parasite. By using seven neoglycoconjugates, we probed the cellular capacity to specifically bind carbohydrate ligands. Similar to Gal-3, an up-regulation was noted with respect to sites specific for Man and alpha-GalNAc, respectively, revealing that infection-dependent changes are not confined to Gal-3-dependent parameters. Considered together, these data document for the first time that a parasitic infection can modulate both in vivo and in vitro the expression of Gal-3 and of ligands for this lectin in DCs with functional consequences on their capacities of adhesion and migration. These results suggest a new immunomodulatory property of T. cruzi.
我们研究了病原体诱导的炎症反应对克氏锥虫感染小鼠脾脏树突状细胞（DCs）及其半乳糖凝集素-3（Gal-3）表达的影响。我们使用标记的半乳糖凝集素作为探针，测定了凝集素的表达以及配体的呈递情况。通过逆转录聚合酶链反应、蛋白质免疫印迹分析、定量糖细胞化学和计算机辅助定量显微镜技术，我们证明，在感染小鼠的脾脏树突状细胞中，Gal-3及其特异性配体的表达显著上调，并且与Gal-3包被的基质的黏附性增加。在克氏锥虫感染的D2SC-1细胞中，Gal-3的表达也增强。为了评估对迁移的影响，我们不得不专门使用D2SC-1细胞，因为脾脏树突状细胞迅速丧失了黏附于基质的能力。当基质用Gal-3包被时，感染的和经中药处理的D2SC-1细胞的迁移减少。当D2SC-1细胞与抗Gal-3反义寡核苷酸孵育时，其Gal-3的表达降低，但对寄生虫的细胞侵袭没有影响。通过使用七种新糖缀合物，我们检测了细胞特异性结合碳水化合物配体的能力。与Gal-3类似，分别观察到对甘露糖和α-N-乙酰半乳糖胺特异性位点的上调，这表明感染依赖性变化并不局限于Gal-3依赖性参数。综合考虑，这些数据首次证明寄生虫感染可以在体内和体外调节DCs中Gal-3及其凝集素配体的表达，并对其黏附和迁移能力产生功能影响。这些结果提示了克氏锥虫一种新的免疫调节特性。
