The Rabbit Genome, Immune System and Ig Genetics

家兔是研究多种传染病的唯一或最好的动物模型。2008年7月，在布罗德研究所、麻省理工学院和哈佛完成了7倍覆盖率的兔基因组测序。2009年8月3日，将深度覆盖的组装提交至GenBank（登录号AAGW 02000000）。我们已经能够在不完整的2x跟踪存档和组装沿着的7 x跟踪存档中找到并使用的序列已经被证明对各种研究有用。现在有两个网站：第一个是NIAID关于免疫学和传染病中的兔子的网站（http：www3.niaid.nih.gov/research/resources/ri/）。该网站提供了2005年NIAID举办的人类传染病兔子模型研讨会的摘要。第二个网站是由国家生物技术信息中心（NCBI）维护的网站，http://www.ncbi.nlm.nih.gov/projects/genome/guide/rabbit/.Rabbits，b 9同种异型已用于需要高度特异性高亲和力抗体的各种免疫项目。HIV-1的gp 41的保守的N-七肽重复（NHR）区域形成卷曲螺旋，其是融合反应的前体。从免疫噬菌体展示文库中分离HIV-1中和性单克隆抗体，所述噬菌体展示文库来自用gp 41的NHR区域的NHR卷曲螺旋的模拟物（N35 CCG-N13）免疫的b 9同种异型兔。来自一只兔的三个回收的中和scFv的VDJ序列是克隆相关的，并且具有由很少表达的VH基因编码的重链可变区（VH）（x32）（纳尔逊et al.2008）J 9兔的免疫还产生了对NgRl和NgR 2特异性的mAb，其用于表征CNS髓磷脂抑制的新型拮抗剂（Robak et al，2009）。我们已发表的开发SLE兔模型的工作（Puliyath et al，2008，以及其中的参考文献）引起了人们对使用选择性繁殖的兔产生自身抗体的兴趣，这些抗体用于旨在开发新疫苗以引发广泛中和的抗HIV应答的研究。据推测，由于潜在的自身抗体活性，编码能够广泛中和不同HIV毒株的抗体的B细胞在B细胞发育期间被选择。还已知BAFF介导的未成熟B细胞的存活有助于自身反应性B细胞的存活和成熟。几个小组将或目前正在使用我们以前研究中报告的纯种兔的亲属研究对可能引起广泛中和HIV抗体的各种抗原的反应，因为它们更可能产生自身抗体。正在进行的一项研究计划还包括使用我们描述的重组兔BAFF（Yang et al. 2009）。在最近的研究中，筛选了从免疫的兔产生的b 9同种异型的嵌合兔/人Fab文库，以鉴定对HIV-1的Rev蛋白的天然表位具有高亲和力和特异性的嵌合兔/人Fab。我们假设嵌合兔/人Fab可以促进共结晶。通过与独特的表位结合，Fab显示出有效地溶解Rev聚合物。Fab/Rev复合物被纯化、表征和结晶。作为晶体伴侣的能力是嵌合兔/人Fab的广泛用途的新应用。从b 9兔免疫中选择的mAb也具有由不寻常的VH基因编码的重链可变区。(Ms在制备S中。斯塔尔，N.瓦茨，P.温菲尔德，等。NIAMS，R法师，NIAID，C. Rader NCI）。

Rabbits are the only or best animal model for several infectious diseases. The sequence of the rabbit genome at 7x coverage was completed in July 2008 at Broad Institute MIT and Harvard. Assembly of the deep coverage was submitted to GenBank (accession AAGW02000000) on August 3, 2009. The sequences we have been able to find and use in the incomplete 2x trace archive and assembly along with the 7x trace archive have already proven useful for various studies. Two websites are now available:the first is by NIAID on Rabbit in Immunology & Infectious Disease(http://www3.niaid.nih.gov/research/resources/ri/. This site offers a summary of an NIAID workshop on Rabbit Models of Human Infectious diseases held in 2005. The second site is one maintained by the National Center for Biotechnology Information (NCBI), http://www.ncbi.nlm.nih.gov/projects/genome/guide/rabbit/.Rabbits of the b9 allotype have been used in a variety of immunization projects where highly specific high affinity antibodies are desired. The conserved N-heptad repeat (NHR) region of gp41of HIV-1 forms a coiled-coil that is a precursor to the fusion reaction. HIV-1-neutralizing monoclonal antibodies were isolated from immune phage display libraries from rabbits of b9 allotype immunized with a mimetic of the NHR coiled-coil, (N35CCG-N13) of the NHR region of gp41. The VDJ sequences of the three recovered neutralizing scFv from one rabbit were clonally related and had a heavy chain variable region (VH) encoded by a rarely expressed VH gene (x32) (Nelson et al. 2008).Immunization of b9 rabbits also led to mAbs specific for NgR1 and NgR2 that were used for characterization of a novel antagonist of CNS myelin inhibition (Robak et al, 2009). Our published work to develop a rabbit model of SLE (Puliyath et al, 2008, and reference therein) has a raised interest in the use of rabbits selectively bred to produce autoantibodies for studies aimed at development of new vaccines to elicit broadly neutralizing anti-HIV responses. It has been hypothesized that B-cells encoding antibodies capable of broadly neutralizing different HIV strains are selected against during B-cell development due to potential autoantibody activity. It is also known that BAFF-mediated survival of immature B cells contributes to the survival and maturation of autoreactive B cells. The responses to various antigens that may elicit broadly neutralizing antibodies to HIV will be or are currently being investigated by several groups using relatives of the pedigreed rabbits reported in our previous studies because they are more likely to produce autoantibodies. The plan of one study in progress also includes use of recombinant rabbit BAFF that we described (Yang et al. 2009). In more recent studies, a chimeric rabbit/human Fab library generated from immunized rabbits, of b9 allotype was screened to identify chimeric rabbit/human Fab of high affinity and specificity for a native epitope of the Rev protein of HIV-1. We hypothesized that chimeric rabbit/human Fab could facilitate co-crystallization. By binding to a unique epitope, the Fab was shown to potently dissolve Rev polymers. Fab/Rev complexes were purified, characterized and crystallized. The ability to serve as crystal chaperones is a new application of broad utility for chimeric rabbit/human Fab. The selected mAb from b9 rabbit immunization also had a heavy chain variable region encoded by an unusual VH gene. (Ms in preparation S. Stahl, N.Watts, P.Wingfield, et al. PEB. NIAMS, R Mage, NIAID, C. Rader NCI).