A XENOPUS LAEVIS RESEARCH RESOURCE FOR IMMUNOBIOLOGY

描述（申请人提供）：非洲爪蟾是研究免疫系统系统发育和个体发生的最佳变温脊椎动物模型之一。X. laevis与哺乳动物的进化距离允许区分物种特异性适应与更保守的免疫系统特征。X. laevis提供了一种独特的、通用的、非哺乳动物模型，用于研究MHC限制和无限制识别、个体发生和系统发生以及针对肿瘤、病毒和细菌的体液和细胞介导免疫。特别是，在变态过程中MHC I类分子的发育调节获得，以及在I类和其他成人特异性抗原表达之前可以通过实验操作胚胎和幼虫的容易性，使人们能够解决有关MHC限制，自身免疫和自我耐受性发展的问题，这些问题在其他动物模型中不易研究。几十年来，对X. laevis的研究已经产生了许多宝贵的研究工具，包括mhc定义的等基因克隆和动物近交系、转基因系、细胞系、单克隆抗体和cDNA探针，这些工具需要保存、丰富并提供给科学界。因此，这一更新提案的广泛目标是维护、促进和进一步发展X. laevis作为生物医学研究的重要模型，特别是免疫学研究。与最初的建议一样，我们提出了两个主要目标：(1)通过继续保持和提高资源的性能和质量，维护、改善和宣传我们的X. laevis设施；应要求提供非市售动物和试剂；通过协助、培训和告知科学家和学生有关X. laevis的信息；通过网站传播信息、做广告和与科学界互动。(2)开发新的实验动物、方法和试剂，生产新的MHC和小h抗原等基因克隆集合；采用转基因技术，产生转基因基因，即等基因克隆，在淋巴组织（如胸腺、脾脏）或白细胞中表达荧光报告基因；利用转基因技术通过RNA干扰进行体内敲低，揭示免疫相关基因的功能；并通过产生新的X. laevis特异性抗体（Abs）来识别免疫相关分子。

DESCRIPTION (provided by applicant): Xenopus laevis is one of the best ectothermic vertebrate models for studying the phylogeny and ontogeny of the immune system. The evolutionary distance of X. laevis from mammals permits distinguishing species-specific adaptations from more conserved features of the immune system. X. laevis, provides a unique, versatile, non-mammalian model with which to study humoral and cell-mediated immunity in the context of MHC restricted and unrestricted recognition, ontogeny, and phylogeny, and against tumors, viruses, and bacteria. In particular, the developmentally regulated acquisition of MHC class I molecules during metamorphosis and the ease with which one can experimentally manipulate embryos and larvae prior to expression of class I and other adult-specific antigens allows one to address questions about MHC restriction, autoimmunity, and the development of self-tolerance that can not be easily studied in other animal models. Studies with X. laevis over several decades have resulted in the generation of many invaluable research tools including MHC-defined isogenetic clones and inbred strains of animals, transgenic lines, cell lines, monoclonal antibodies, and cDNA probes that need to be preserved, enriched and made available to the scientific community. The broad objective of this renewal proposal, therefore, is to safeguard, promote and further develop X. laevis as an important model for biomedical research in general, and immunology, in particular. As in the original proposal, two major aims are proposed: (1) Maintenance, improvement and advertisement of our X. laevis facility by continuing to maintain and improve the performance and quality of our resource; by providing animals, not commercially available, and reagents upon request; by assisting, training, and informing scientists and students about X. laevis; and by disseminating information, advertising and interacting with the scientific community through a web site. (2) Development of new experimental animals, methodologies, and reagents by producing a new collection of isogenetic clones MHC and minor-H-antigen defined; by adapting transgenesis techniques and generating transgenic, which are isogenetic clones, expressing fluorescent reporter genes in lymphoid tissues (e.g., thymus, spleen) or leukocytes; by developing in vivo knockdown by RNA interference using transgenesis to reveal the function of immunologically-relevant genes; and by generating new X. laevis-specific antibodies (Abs) recognizing immunologically-relevant molecules. RELEVANCE: The overall objective of this renewal application is to safeguard and promote the frog Xenopus laevis, as an important non-mammalian comparative model for biomedical research in general, and immunology, in particular. We are proposing to continue the maintenance and the development of this unique non-mammalian resource facility for biomedical research for the benefit of the whole scientific community.