Ramírez-Gómez Francisco, Ortiz-Pineda Pablo A, Rivera-Cardona Gabriela, García-Arrarás José E
Department of Biology, University of Puerto Rico, Río Piedras, San Juan, Puerto Rico.
PLoS One. 2009 Jul 8;4(7):e6178. doi: 10.1371/journal.pone.0006178.
Metazoan immunity is mainly associated with specialized cells that are directly involved with the immune response. Nevertheless, both in vertebrates and invertebrates other organs might respond to immune activation and participate either directly or indirectly in the ongoing immune process. However, most of what is known about invertebrate immunity has been restricted to immune effector cells and little information is available on the immune responses of other tissues or organs. We now focus on the immune reactions of the intestinal tissue of an echinoderm. Our study employs a non-conventional model, the echinoderm Holothuria glaberrima, to identify intestinal molecules expressed after an immune challenge presented by an intra-coelomic injection of lipopolysaccharides (LPS). The expression profiles of intestinal genes expressed differentially between LPS-injected animals and control sea water-injected animals were determined using a custom-made Agilent microarray with 7209 sea cucumber intestinal ESTs. Fifty (50) unique sequences were found to be differentially expressed in the intestine of LPS-treated sea cucumbers. Seven (7) of these sequences represented homologues of known proteins, while the remaining (43) had no significant similarity with any protein, EST or RNA database. The known sequences corresponded to cytoskeletal proteins (Actin and alpha-actinin), metabolic enzymes (GAPDH, Ahcy and Gnmt), metal ion transport/metabolism (major yolk protein) and defense/recognition (fibrinogen-like protein). The expression pattern of 11 genes was validated using semi-quantitative RT-PCR. Nine of these corroborated the microarray results and the remaining two showed a similar trend but without statistical significance. Our results show some of the molecular events by which the holothurian intestine responds to an immune challenge and provide important information to the study of the evolution of the immune response.
后生动物免疫主要与直接参与免疫反应的特化细胞相关。然而,在脊椎动物和无脊椎动物中,其他器官也可能对免疫激活作出反应,并直接或间接参与正在进行的免疫过程。然而,目前关于无脊椎动物免疫的大部分知识都局限于免疫效应细胞,关于其他组织或器官的免疫反应的信息很少。我们现在关注一种棘皮动物肠道组织的免疫反应。我们的研究采用了一种非传统模型,即棘皮动物光滑海参,以鉴定在体腔注射脂多糖(LPS)引发免疫挑战后肠道中表达的分子。使用定制的安捷伦微阵列,其中包含7209个海参肠道EST,确定了LPS注射动物和对照海水注射动物之间差异表达的肠道基因的表达谱。发现50个独特序列在LPS处理的海参肠道中差异表达。其中7个序列代表已知蛋白质的同源物,其余43个与任何蛋白质、EST或RNA数据库均无显著相似性。已知序列对应于细胞骨架蛋白(肌动蛋白和α-辅肌动蛋白)、代谢酶(甘油醛-3-磷酸脱氢酶、腺苷高半胱氨酸酶和甘氨酸-N-甲基转移酶)、金属离子运输/代谢(主要卵黄蛋白)和防御/识别(纤维蛋白原样蛋白)。使用半定量RT-PCR验证了11个基因的表达模式。其中9个证实了微阵列结果,其余2个显示出类似趋势,但无统计学意义。我们的结果显示了海参肠道对免疫挑战作出反应的一些分子事件,并为免疫反应进化的研究提供了重要信息。