Rombout J H W M, van der Tuin S J L, Yang G, Schopman N, Mroczek A, Hermsen T, Taverne-Thiele J J
Cell Biology and Immunology Group, Department of Animal Sciences, Wageningen University, PO Box 338, 6700 AH Wageningen, The Netherlands.
Fish Shellfish Immunol. 2008 May;24(5):620-8. doi: 10.1016/j.fsi.2008.01.016. Epub 2008 Feb 8.
The mucosal immune system seems to be an important defence mechanism for fish but the binding of IgM in mucosal organs is poorly described in fish. In this study the gene encoding the polymeric Immunoglobulin Receptor (pIgR) in carp has been isolated and sequenced from a liver cDNA-library and aligned with other species. The pIgR of carp consists of 2 Ig domains, a transmembrane and an intracellular region, together 327 amino acids. In situ hybridisations with sense and anti-sense DIG-labelled pIgR RNA probes were performed on liver, gut and skin of common carp (Cyprinus carpio L.) and in these organs only anti-sense probes were found to hybridise. In liver the majority of hepatocytes was stained around the nucleus. In gut and skin, staining could be detected around the nucleus of the epithelial cells, but in gut also a subpopulation of lymphoid cells was stained in epithelium and lamina propria. The specific in situ hybridisation of the epithelia and hepatocytes coincides with the in situ binding of FITC-labelled carp IgM to the same cells. RT-PCR results indicate the expression of the pIgR gene in all lymphoid organs of carp, but not in muscle. Macrophages/neutrophils enriched by adherence or sorted B cells (MACS) did not show expression of the pIgR gene and are excluded as the pIgR expressing lymphoid cells in the intestine. The relevance of pIgR staining and gene expression in mucosal organs is discussed.
黏膜免疫系统似乎是鱼类重要的防御机制,但鱼类黏膜器官中IgM的结合情况鲜有描述。在本研究中,从鲤鱼肝脏cDNA文库中分离并测序了编码多聚免疫球蛋白受体(pIgR)的基因,并与其他物种进行了比对。鲤鱼的pIgR由2个Ig结构域、1个跨膜区和1个胞内区组成,共327个氨基酸。用有义链和反义链地高辛标记的pIgR RNA探针,对鲤鱼(Cyprinus carpio L.)的肝脏、肠道和皮肤进行原位杂交,结果发现只有反义探针能杂交。在肝脏中,大多数肝细胞在细胞核周围被染色。在肠道和皮肤中,上皮细胞核周围可检测到染色,但在肠道中,上皮和固有层中的一部分淋巴细胞也被染色。上皮细胞和肝细胞的特异性原位杂交与异硫氰酸荧光素标记的鲤鱼IgM在相同细胞上的原位结合相一致。逆转录聚合酶链反应(RT-PCR)结果表明,pIgR基因在鲤鱼所有淋巴器官中表达,但在肌肉中不表达。通过贴壁富集的巨噬细胞/中性粒细胞或分选的B细胞(磁珠分选)未显示pIgR基因表达,因此排除其为肠道中表达pIgR的淋巴细胞。文中讨论了pIgR染色和基因表达在黏膜器官中的相关性。