Structure, function and disease susceptibility of bovine MHC class II molecules

牛MHC II类分子的结构、功能和疾病易感性

• 批准号: 11460140
• 负责人: AIDA Yoko
• 金额: $ 10.3万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460140/
• 关键词: BoLA class II gene; DNA typing; PCA-Sequence-Based Typing; BoLA-DRB3 exon 2; BLV-infected cattle; Resistance allele; Polymorphism; Sheep

The bovine leukocyte antigen (BoLA)class II genes code for highly polymorphic transmembrane glycoproteins that present antigenic peptides to helper T cells. The BoLA-DRB3 is thought to be the most functionally significant as it is actively transcribed and in excess of 102 alleles have been characterized for this locus. First, we developed the technique of polymerase chain reaction-sequence based typing(PCR-SBT)which can assign DRB3 alleles in DNA sequence level and would allow fast screening of large numbers of animals. Next, we designed to determine the nucleotide sequences of exon 2 of BoLA-DRB3 alleles in a total of 419 animalks from four distinct cattle breeds such as Japanese Black. Japanese Shorthorn Holstein and Jersey by our PCR-SBT method, and the allele frequencies and the phylogenetic relationship of BoLA-DRB3 alleles and the genetic background of these populations are discussed.The nucleotide sequencing of exon 2 of the BoLA-DRB3 gene, which is highly polymorphic, of 81 bov … More ine leukemia virus(BLV)-infected animals with 3 independent stages such as an aleukemic healthy, persistent lymphocytosis(PL)and lymphoma was determined by PCR-sequenced-based typing. We identified 23 distinct BoLA-DR83 alleles, including 3 new alleles. The population of healthy cattle positive for BoLA-DRB3^*14011 was higher than the proportion-bearing individuals in 200 control cases that were positive for the same allele. By contrast, the BoLA-DRB3^*1601 alleles was found most frequently in cattle with PL and lymphoma. Sequence analysis revealed that, approximately 56 % of 43 BLV-infected but healthy cattle carry at least one BoLA-DRB3 allele encoding Arg^<71> of Lys^<71>, Glu^<74>, Arg^<77> and Val^<78> of β1 domain of DR molecule, which suggested that alleles encoding the KERV and RERV motifs might protect against tumor development. By contrast, approximately 70% of 23 BLV-infected cattle with lymphoma carry two alleles encoding Ala^<74>, Thr^<77> and Tyr^<78>, indicating that the ATY/ATY genotype might be associated with susceptibility to lymphoma. Such, these results suggest that the existence of alleles associated with resistance and susceptibility to BLV-induced leukemogenesis.Next, to confirm the association between polymorphism of MHC class II gene and lymphoma, we infected 28 sheep with BLV.Among the 16 sheep with lymphoma, 38% carried specific alleles of the ovine MHC(OLA)-DRB1 gene encoding Arg-Lys(RK)at residues 70 and 71 as compared to 83% among the 12 sheep at the asymptomatic stage. Furthermore, we immunized sheep carrying either a RK/XX genotype or a XX/XX genotype with a synthetic antigenic peptides of T-helper, T-cytotoxic, and B-cell epitopes of BLV envelope glycoprotein gp 51, and two weeks after the last immunization, all sheep were challenged with blood from a BLV-infected sheep. Sheep carrying resistant allele showed high immunoresponse against the mixed peptides and did not proceeded to leukemia. Less

牛白细胞抗原（BoLA）II类基因编码高度多态性的跨膜糖蛋白，将抗原肽呈递给辅助T细胞。BoLA-DRB 3被认为是功能上最重要的，因为它是活跃转录的，并且超过102个等位基因已经被表征为该基因座。首先，我们开发了聚合酶链反应-基于序列的分型（PCR-SBT）技术，该技术可以在DNA序列水平上分配DRB 3等位基因，并允许快速筛选大量动物。接下来，我们设计确定BoLA-DRB 3等位基因外显子2的核苷酸序列，共419头动物，从四个不同的牛品种，如日本黑牛。采用PCR-SBT方法对日本短角牛、荷斯坦牛和泽西牛的BoLA-DRB 3基因进行了遗传分析，探讨了BoLA-DRB 3基因的等位基因频率、系统发育关系和遗传背景，并对81头牛BoLA-DRB 3基因外显子2进行了核苷酸序列测定，结果表明，BoLA-DRB 3基因在日本短角牛、荷斯坦牛和泽西牛中具有高度多态性。 ...更多信息 用PCR-测序分型法对白血病病毒（BLV）感染的动物进行了3个独立阶段的分型，如无白细胞血症的健康动物、持续性淋巴细胞增多症（PL）和淋巴瘤。我们鉴定了23个不同的BoLA-DR 83等位基因，包括3个新的等位基因。BoLA-DRB 3 *14011阳性的健康牛群体高于200个对照病例中相同等位基因阳性的携带个体的比例。相比之下，BoLA-DRB 3 ^*1601等位基因在PL和淋巴瘤牛中最常见。序列分析表明，在43头BLV感染但健康的牛中，约56%携带至少一个编码DR分子β1结构域的Arg <71>^、<71>Glu^<74>、Arg^<77>和瓦尔^<78>的BoLA-DRB 3等位基因，这表明编码KERV和RERV基序的等位基因可能对肿瘤的发生具有保护作用。相比之下，约70%的23头BLV感染的患有淋巴瘤的牛携带编码Ala^<74>、Thr^<77>和Tyr^<78>的两个等位基因，表明ATY/ATY基因型可能与淋巴瘤易感性相关。因此，这些结果表明，存在与BLV诱导的白血病抗性和易感性相关的等位基因。接下来，为了证实MHC II类基因多态性与淋巴瘤的关联，我们用BLV感染了28只绵羊。38%的绵羊携带编码Arg-Lys（RK）的MHC（奥拉）-DRB 1基因第70和71位残基的特异等位基因，而12只无症状期绵羊携带该基因的比例为83%。此外，我们用BLV包膜糖蛋白gp 51的T辅助细胞、T细胞毒性细胞和B细胞表位的合成抗原肽免疫携带RK/XX基因型或XX/XX基因型的绵羊，最后一次免疫后两周，所有绵羊用来自BLV感染绵羊的血液进行攻击。携带抗性等位基因的绵羊对混合肽表现出高的免疫应答，并且没有发生白血病。少

项目成果

Tajima S.: "The region between amino acids 245 and 265 of the Tax protein of bovine leukemia virus (BLV) restricts transactivation not only via the BLV enhancer but also via other retroviral enhancer."J. Virology. 74. 10939-10949 (2000)

Tajima S.：“牛白血病病毒 (BLV) Tax 蛋白的氨基酸 245 和 265 之间的区域不仅通过 BLV 增强子限制反式激活，而且还通过其他逆转录病毒增强子限制反式激活。”

間陽子: "免疫応答遺伝子の多型解析を用いた抗病性ウシ作出への基礎的研究"畜産技術. 17. 24-27 (1999)

Yoko Hazama：“利用免疫反应基因多态性分析生产抗病牛的基础研究”《牲畜技术》17. 24-27 (1999)。

Hishinuma M., Kadohira M., Patel O.V., Zulu V., Okada K., Koie H.and Sekine J.: "Investigation of ovarian follicular population in indigenous barotse cows of Zambis"Bull. Fac. Agric., Tottori Univ.. 53. 103-107 (2000)

Hishinuma M.、Kadohira M.、Patel O.V.、Zulu V.、Okada K.、Koie H. 和 Sekine J.：“赞比斯本土巴罗策牛卵巢卵泡数量的调查”公牛。

間陽子: "免疫遺伝学"獣医遺伝学入門(鈴木勝士監訳、学窓社). 160-172 (1999)

Yoko Hazama：“免疫遗传学”兽医遗传学简介（Katsushi Suzuki 监督翻译，Gakusosha）160-172（1999）。

Inabe K., Nishizawa M,Tajima S,Ikuta K.and Aida Y.: "The YXXL sequences of a transmembrane protein bovine leukemia virus are required for viral entry and incorporation of viral envelope protein into virion."J.Virology. 73. 1293-1301 (1999)

Inabe K.、Nishizawa M、Tajima S、Ikuta K. 和 Aida Y.：“跨膜蛋白牛白血病病毒的 YXXL 序列是病毒进入并将病毒包膜蛋白掺入病毒粒子所必需的。”J.Virology。