Adams J E, Peña M T, Gillis T P, Williams D L, Adams L B, Truman R W
Laboratory Research Branch National Hansen's Disease Programs, Louisiana State University, School of Veterinary Medicine, Skip Bertman Drive, Baton Rouge, LA 70803, USA.
Cytokine. 2005 Dec 7;32(5):219-25. doi: 10.1016/j.cyto.2005.09.011. Epub 2005 Dec 9.
The nine-banded armadillo (Dasypus novemcinctus) is the only immunologically intact animal that regularly develops lepromatous-type leprosy when inoculated with Mycobacterium leprae. However, the ability to exploit this model for understanding the pathogenesis of leprosy has been limited by a lack of suitable immunological reagents. Recently, efforts began to sequence the entire armadillo genome, and this sequence information will help make possible the development of a wide array of new immunological reagents suitable for use with armadillos. Using the available sequence data, a region of high homology to interleukin-2 of other mammals was identified. Primers were designed to amplify the coding region corresponding to the mature peptide and its exact sequence was confirmed. cDNA was made from ConA-stimulated armadillo PBMC. The amplified coding region was sub-cloned into a pET expression vector and transformed into Escherichia coli for over-expression. The subsequent product was characterized by SDS-PAGE and bioassays. Tritiated thymidine incorporation by CTLL-2 and armadillo lymphoblasts confirmed functionality of the recombinant product. The advent of the D. novemcinctus genome sequence and subsequent generation of immunological tools will assist in advancing the armadillo as a translational model for leprosy.
九带犰狳（Dasypus novemcinctus）是唯一一种在接种麻风分枝杆菌后会定期发展为瘤型麻风的免疫健全动物。然而，由于缺乏合适的免疫试剂，利用该模型来理解麻风病发病机制的能力受到了限制。最近，人们开始对犰狳的整个基因组进行测序，这些序列信息将有助于开发一系列适用于犰狳的新型免疫试剂。利用现有的序列数据，鉴定出了与其他哺乳动物白细胞介素-2具有高度同源性的区域。设计引物以扩增对应于成熟肽的编码区，并确认其确切序列。从刀豆蛋白A刺激的犰狳外周血单核细胞制备cDNA。将扩增的编码区亚克隆到pET表达载体中，并转化到大肠杆菌中进行过表达。随后的产物通过SDS-PAGE和生物测定进行表征。CTLL-2和犰狳淋巴母细胞对氚标记胸腺嘧啶核苷的掺入证实了重组产物的功能。九带犰狳基因组序列的出现以及随后免疫工具的产生将有助于推动犰狳成为麻风病的转化模型。
