Skin Grafting in Xenopus laevis: A Technique for Assessing Development and Immunological Disparity
Skin Grafting in Xenopus laevis: A Technique for Assessing Development and Immunological Disparity
复制标题
非洲爪蟾的皮肤移植：一种评估发育和免疫差异的技术
DOI：
10.1101/pdb.prot099788
复制
发表时间：
2018
影响因子：
--
通讯作者：
Izutsu Y.
中科院分区：
文献类型：
--
作者：
篠田 雄大;染谷 友美;Izutsu Y.
两栖动物非洲爪蟾（Xenopus laevis）的皮肤移植不仅用于检测由次要H抗原或MHC单倍型不同的同种异体抗原，而且还用于检测变态过程中个体特异性抗原（包括新出现的成虫特异性和消失的幼虫特异性）。为了了解针对幼虫和/或成人特异性抗原的同种异体耐受或免疫反应的机制，一个完整的MHC纯合子近交系是最合适的实验模型。这里使用的是在日本建立的近交系J。自交的J株具有完全的组织相容性，在同一株成年蛙中没有出现移植皮肤排斥反应，2016年通过基因组序列分析再次证实了其纯合性。因此，J毒株使免疫学家和胚胎学家能够了解发育过程中的进化过程以及免疫事件和组织重塑机制。此外，我们的实验室还提供了J菌株与gfp标记的转基因株系之间的F1杂交株，可作为长期细胞跟踪的模型。本协议解释了皮肤移植的方法，以确定宿主和供体之间的免疫差异。在早期胚胎发生和从幼虫到成虫的完全发育过程中，也可以追踪宿主的细胞和组织命运，这在其他物种中是极其困难的。
Skin grafting in the amphibian Xenopus laevis has been used to detect not only allogeneic antigens that differ by minor H antigens or by one MHC haplotype, but also to detect ontogeny-specific antigens (including both emerging adult-and disappearing larval-specific) during metamorphosis. To understand the mechanisms underlying allogeneic tolerance or immune responses against larval-and/or adult-specific antigens, a complete MHC homozygous, inbred strain is the most appropriate experimental model. The inbred J strain established in Japan is used here. Owing to complete histocompatibility, the inbred J strain shows no grafted skin rejection among the same strain of adult frogs, and its genuine homozygosity was reconfirmed by genomic sequence analysis in 2016. Therefore, the J strain enables immunologists and embryologists to understand evolutionary processes as well as immunological events and tissue remodeling mechanisms present during development. Furthermore, an F1 hybrid between the J strain and a GFP-labeled transgenic line is available from our laboratory and can be used as a model for long-term cell tracking. This protocol explains the methodology for skin grafting in X. laevis to determine immunological discrepancies between the host and donor. It is also possible to trace cell and tissue fates in the hosts during early embryogenesis and during complete development from larvae to adults, which is extremely difficult to perform using other species.