HLA-A and B polymorphisms predate the divergence of humans and chimpanzees

Major histocompatibility complex (MHC) glycoproteins bind processed fragments of proteins and present them to the receptors of T lymphocytes. The extraordinary polymorphism of class I MHC molecules in man (HLA-A, B and C) and mouse (H-2 K, D and L) poses many questions concerning their diversification and evolution. Comparison of allelic sequences within a species suggests diversity is generated by the assortment of point mutations into varied combinations by mechanisms of recombination and gene conversion. We have now compared class I MHC alleles in two closely related species: humans (Homo sapiens) and chimpanzees (Pan troglodytes). Chimpanzee homologues of HLA-A, HLA-B and a non-classical gene have been identified. No features distinguishing human and chimpanzee alleles could be found. Individual HLA-A or B alleles are more closely related to individual chimpanzee alleles than to other HLA-A or B alleles. These results show that a considerable proportion of contemporary HLA-A and B polymorphism existed before divergence of the chimpanzee and human lines. The stability of the polymorphism indicates that hyper-mutational mechanisms are not necessary to account for HLA-A, B and C diversity.     

MHC polymorphism pre-dating speciation

Two features distinguish the polymorphism of the major histocompatibility complex (MHC) loci from that of other loci: its high diversity and the large genetic distance between MHC alleles. More than 100 alleles exist in natural populations in the mouse at each of the functional class I and class II alleles, all alleles occurring at frequencies that cannot be explained by recurrent mutations. Some of the alleles differ by approximately 70 nucleotides in the coding region alone and some of the products of the allelic genes differ by more than 50 amino acids. It has generally been assumed that these differences accumulated after species inception. Here, we present evidence for an alternative explanation of the origin of MHC polymorphism: a large part of the MHC polymorphism pre-dates speciation and is passed on from species to species. We describe allelic differences that must have arisen before the separation of mice and rats from a common ancestor more than 10 million years ago.     

MHC antigens in urine as olfactory recognition cues

The classical class I antigens of the major histocompatibility complex (MHC) are cell-surface glycoproteins which were originally discovered because they cause rapid rejection of cells or tissues grafted between unrelated individuals. These molecules are encoded by the K, D and L loci of the mouse MHC (and analogous loci in other species) which show extreme species polymorphism and a large number of alleles. In an outbreeding population 3.6 X 10(9) unique MHC class I phenotypes can be encoded by the 100 alleles at each of the K and D loci and the 6 alleles at the L locus. This level of polymorphism ensures that the cells and tissues of each unrelated individual are uniquely identified by their class I membrane-bound antigens. Like other membrane bound proteins, these class I molecules are anchored in the lipid bilayer by a hydrophobic domain encoded by exon 5. However, there have been reports of the occurrence of classical class I molecules in true solution in the blood of humans, mice, and rats. We report here that classical polymorphic class I molecules in normal rats are constitutively excreted in the urine and that untrained rats can distinguish the smell of urine samples taken from normal donors that differ only at the class I MHC locus and therefore excrete different allelomorphs of class I molecules in their urine.     

Evolutionary relationship among Bothriocephalusparasitized in grass carp Ctenopharyngodon idellus (C. et V.), common carp Cyprinus carpio L. and Ma Kou Yu Opsariichthys bidens Günther in China

RAPD was used to analyze the polymorphism of \%Bothriocephalus\% parasitic in grass carp and common carp from Guangdong, Fujian and Gansu and in Ma Kou Yu from Fujian. Though there exist differences in geographical distribution and host specificity between \%Bothriocephalus\% parasitic in grass carp and common carp, RAPD analysis shows high similarity (72.3%-96.3%) in DNA between them, whereas between \%Bothriocephalus\% parasitic in Ma Kou Yu and in grass carp and common carp low similarity (24%-28.2%) was noticed. Hence, \%Bothriocephalus\% parasitic in Ma Kou Yu and \%Bothriocephalus\% parasitic in grass carp and common carp should be regarded as separate species.     

Evolutionary relationship amongBothriocephalus parasitized in grass carpCtenopharyngodon idellus (C. et V.), common carpCytpinus carpio L. and Ma Kou YuOpariichthys bidens Günther in China

RAPD was used to analyze the polymorphism ofBothriocephalus parasitic in grass carp and common carp from Guangdong, Fujian and Gansu and in Ma Kou Yu from Fujian. Though there exist differences in geographical distribution and host specificity betweenBothriocephulus parasitic in grass carp and common carp, RAPD analysis shows high similarity (72.3%–96.3%) in DNA between them, whereas betweenBothriocephalus parasitic in Ma Kou Yu and in grass carp and common carp low similarity (24%-28.2%) was noticed. Hence,Bothriocephalus parasitic in Ma Kou Yu andBothriocephalus parasitic in grass carp and common carp should be regarded as separate species.     

草鱼原肌球蛋白基因的克隆及其组织表达

为揭示原肌球蛋白基因在草鱼肌肉中的作用,利用RT-PCR和RACE技术克隆获得了草鱼原肌球蛋白基因c DNA,并对该基因在普通草鱼和脆肉鲩不同组织中的表达情况进行研究分析。结果表明原肌球蛋白基因c DNA全长序列为1 705 bp,包含387 bp的5′UTR序列,1 307 bp的3′UTR序列和855 bp开放阅读框(ORF)。其ORF编码284个氨基酸。系统进化分析表明普通草鱼与斑马鱼、墨西哥脂鲤的原肌球蛋白基因核苷酸同源性分别是93%和87%,氨基酸同源性分别是96%和93%。在聚类上普通草鱼原肌球蛋白基因与其他鲤科鱼类同源性较高,表明亲缘关系最近,与传统分类相一致。Real time-PCR结果表明原肌球蛋白基因在所检测的普通草鱼和脆肉鲩7个组织中均有表达,原肌球蛋白基因在普通草鱼腹肌中表达最高,其次为前肠。原肌球蛋白基因在脆肉鲩腹肌中的表达低于普通草鱼,而脆肉鲩中肌肉、肝脏、肾脏、前肠、后肠中原肌球蛋白基因表达量大于普通草鱼相对应组织,但差异不显著。     

Truncation variants of peptides isolated from MHC class II molecules suggest sequence motifs.

T cells recognize foreign protein antigens in the form of peptide fragments bound tightly to the outer aspect of molecules encoded by the major histocompatibility complex (MHC). Most of the amino-acid differences that distinguish MHC allelic variants line the peptide-binding cleft, and different allelic forms of MHC molecules bind distinct peptides. It has been demonstrated that peptide-binding to MHC class I involves anchor residues in certain positions and that antigenic peptides associated with MHC class I exhibit allele-specific structural motifs. We have previously reported an analysis of MHC class II-associated peptide sequences. Here we extend this analysis and show that certain amino-acid residues occur at particular positions in the sequence of peptides binding to a given MHC class II molecule. These sequence motifs require the amino terminus to be shifted one or two positions to obtain alignment; such shifts occur naturally for a single peptide sequence without qualitatively altering CD4 T-cell recognition.     

草鱼基因组随机扩增多态性引物及多态性位点的筛选

为了建立草鱼基因组DNA多态性分析的指标体系,应用RAPD技术,从180条10碱基随机引物中筛选出了31条能扩增出多态性DNA片段的引物。用这31条多态性引物共扩增出了327条重复性好、带型清晰、分辨率高的谱带。扩增产物的片段大小范围在400—2000bp之间。单一引物扩增条带为5—14条。用这些多态性引物在草鱼基因组DNA中检测到了93个多态性位点,并对这些多态性位点上的等位基因频率进行了统计分析。这31条多态性引物和所检测到的93个多态性位点初步为草鱼基因组的多态性分析提供了可靠的分析指标体系。     

Polymorphism of human Ia antigens: gene conversion between two DR beta loci results in a new HLA-D/DR specificity

The polymorphic HLA-DR beta-chains are encoded within the human major histocompatibility complex (MHC) by multiple loci resulting from gene duplications. Certain DR haplotypes can be grouped into families based on shared structural factors. We have studied the molecular basis of HLA-DR polymorphism within such a group which includes the haplotypes DR3, DR5 and DRw6. Molecular mapping of the DR beta-chain region allows true allelic comparisons of the two expressed DR beta-chain loci, DR beta I and DR beta III. At the more polymorphic locus, DR beta I, the allelic differences are clustered and may result from gene conversion events over very short distances. The gene encoding the HLA-DR3/Dw3 specificity has been generated by a gene conversion involving the DR beta I and the DR beta III loci of the HLA-DRw6/Dw18 haplotype, as recipient and donor gene, respectively. Based on which allele is found at DR beta III, the less polymorphic locus, two groups of haplotypes can be defined: DRw52a and DRw52b. The generation of HLA-DR polymorphism within the DRw52 supertypic group can thus be accounted for by a succession of gene duplication, divergence and gene conversion.     

短时间微流水处理对草鱼鱼肉风味品质的影响

为改善草鱼鱼肉的风味品质,以池塘养殖草鱼为研究对象,采用室内微流水系统处理草鱼0、1、4、7d。采用全二维气相色谱-飞行时间质谱技术,并结合相对气味活度值分析,筛选出不同处理时间草鱼鱼肉中的关键气味物质；同时基于液相色谱-质谱联用技术的代谢组学方法,研究室内微流水处理对草鱼鱼肉滋味成分的影响。结果表明,微流水处理时间对草鱼鱼肉中气味成分的含量和种类有明显影响。随着处理时间的延长,草鱼鱼肉的风味前体物质谷胱甘肽、氧化型谷胱甘肽和多不饱和脂肪酸相对含量增加,关键气味成分中呈青草味的己醛、呈鱼腥味的庚醛、呈绿霉味的(E)-2-戊烯醛和1-庚醇等不良气味物质相对含量降低,滋味物质中呈鲜味和甜味的氨基酸和核苷酸含量累积,呈苦味的氨基酸含量逐渐降低,从而改善了草鱼的肉质口感和风味品质。     

草鱼多肽功能性质及营养价值

研究草鱼多肽的溶解性、起泡性、乳化性、吸湿保湿性和稳定性等功能性质,以及草鱼多肽的氨基酸组成,并评价其营养价值。结果表明草鱼多肽具有良好的功能特性。草鱼多肽粉18种氨基酸总质量分数为90.94%,必需氨基酸占总氨基酸质量分数的41.33%;草鱼多肽的生物价、必需氨基酸指数、营养指数和氨基酸比值系数分别为71.51,76.34,74.81和77.19。草鱼多肽的氨基酸组成平衡、合理,是一种优质的蛋白质资源。     

Beta 2-microglobulin restriction of antigen presentation.

Antigens are generally thought to be recognized by cytotoxic T lymphocytes as peptides in the context of class I major histocompatibility proteins complex, which are heterodimers of heavy chains noncovalently associated with beta 2-microglobulin (beta 2m). The highly polymorphic nature of the heavy chains and their resulting ability to present different sets of peptides has presumably evolved to allow potent immune responses against most pathogens. By contrast, the polymorphism of beta 2m is limited; seven alleles are known in the mouse and only one has been identified in humans. beta 2-Microglobulin was consequently thought to have only structural functions: namely, to ensure correct folding of class I molecules and their transport to the cell surface. Although beta 2m is not implicated directly in the formation of the peptide binding site, we report here that it participates in the selection of MHC class I molecule-associated peptides.     

水分、脂肪含量、蛋白质组成和肌肉纤维的大小对脆肉鲩鱼背肌质构特性的影响

通过对生脆肉鲩鱼背肌和生草鱼背肌的质构特性、水分含量、脂肪含量和蛋白质不同成分的测定,结合扫描电镜技术,研究水分、脂肪含量、蛋白质成分和肌肉纤维的平均大小对质构特性的影响。研究结果表明,脆肉鲩鱼肌肉的硬度、弹性、咀嚼性和回复性均比草鱼高,分别高出11.14%,3.4%, 15.75%和14.27%,但粘聚性比草鱼低16.16%。而脆肉鲩鱼的这些特征与其肌肉中的水分含量、基质蛋白的含量和肌肉纤维的平均直径与密度有关,特别是肌肉纤维的平均直径对脆肉鲩鱼肌肉的硬度影响更大,但是在本研究中脂肪含量对脆肉鲩鱼的硬度影响不大。     

Pattern of nucleotide substitution at major histocompatibility complex class I loci reveals overdominant selection

The major histocompatibility complex (MHC) loci are known to be highly polymorphic in humans, mice and certain other mammals, with heterozygosity as high as 80-90% (ref. 1). Four different hypotheses have been proposed to explain this high degree of polymorphism: (1) a high mutation rate, (2) gene conversion or interlocus genetic exchange, (3) over dominant selection and (4) frequency-dependent selection. In an attempt to establish which of these hypotheses is correct, we examined the pattern of nucleotide substitution between polymorphic alleles in the region of the antigen recognition site (ARS) and other regions of human and mouse class I MHC genes. The results indicate that in ARS the rate of nonsynonymous (amino acid altering) substitution is significantly higher than that of synonymous substitution in both humans and mice, whereas in other regions the reverse is true. This observation, together with a theoretical study and other considerations, supports the hypothesis of overdominant selection (heterozygote advantage).     

137 Cs-γ 辐照对实验红鲫 AFLP 多态性的影响

摘要:目的 观察137 Cs-γ 辐照对实验红鲫 AFLP 的变化,探讨不同剂量137 Cs-γ 辐照对实验红鲫 DNA 多态性的影响。 方法 根据前期实验结果,将实验红鲫 C1HD 系随机分为 5 组,10 尾 / 组,即空白对照组、1 / 16 LD50 、1 / 8 LD50 、1 / 4 LD50 、1 / 2 LD504 个辐照组,设置 1. 94 Gy、3. 88 Gy、7. 76 Gy、15. 53 Gy4 个辐照剂量组和一个空白对照组,用生物辐照仪对实验红鲫进行一次性辐照。 提取辐照前和辐照后的实验红鲫血液 DNA,经 AFLP-PCR 扩增,并进行多态性信息含量( PIC) 、基因杂合度( H) 、香农信息指数( I) 分析。 结果 实验红鲫在137 Cs-γ 辐照处理前的 PIC、H 和 I分别为 45. 20% 、0. 19 和 0. 28,经137 Cs-γ 辐 照 处 理 后 分 别 为 72. 27% 、0. 28 和 0. 41,且 遗 传 学 指 数 升 高。 结 论经137 Cs-γ 辐照处理后,实验红鲫的 AFLP 多态性信息含量增加,基因杂合度升高。 AFLP 标记实验红鲫 C1HD 系可用于监测核辐射污染。     

添加莨菪和疫苗对一龄草鱼促长防病饲养效果的研究

将莨菪和草鱼出血病组织浆疫苗添加于一龄草鱼的日粮中，经４５～６０天的网箱和水泥池饲养结果，口服莨菪和疫苗（含５％ＣＭＣ），既可使草鱼的相对增重率比对照组提高５．７～８２．９％，又可对出血病的免疫保护率达４０～５４．６％。莨菪和疫苗是促进草鱼生长和提高抗病力的有效添加剂。     

Polymorphism in the alpha 3 domain of HLA-A molecules affects binding to CD8

Cytotoxic T lymphocytes (CTL) expressing the CD8 glycoprotein recognize peptide antigens presented by class I major histocompatibility complex (MHC) molecules. This correlation and the absence of CD8 polymorphism led to the hypothesis that CD8 binds to a conserved site of class I MHC molecules. Using a cell-cell binding assay we previously demonstrated specific interaction between human class I MHC (HLA-A,B,C) molecules and CD8. Subsequent analysis of the products of 17 HLA-A,B alleles revealed a natural polymorphism for CD8 binding in the human population. Two molecules, HLA-Aw68.1 and HLA-Aw68.2, which do not bind CD8, have a valine residue at position 245 whereas all other HLA-A,B,C molecules have alanine. Site-directed mutagenesis shows that this single substitution in the alpha 3 domain is responsible for the CD8 binding phenotype and also affects recognition by alloreactive and influenza-specific CTL. Our results indicate that CD8 binds to the alpha 3 domain of class I MHC molecules.     

Tetrameric cell-surface MHC class I molecules.

Purified major histocompatibility complex (MHC) class I molecules have been studied at high resolution by X-ray crystallography; the structure is a complex of a single heavy chain, a beta 2-microglobulin light chain and a tightly bound peptide moiety. We show here that complete MHC class I molecules are post-translationally assembled into tetramers (made up of four heavy chains and four beta 2-microglobulin units) and that this tetrameric species is expressed on the cell surface. The multivalent tetrameric structure of class I molecules can be reconciled with models of T-cell activation that invoke antigen-receptor crosslinking, as opposed to models that depend on an allosteric change.     

血液稀释治疗鲩鱼胆汁中毒的实验研究

新鲜鲩鱼胆汁给动物灌胃可引起多器官损伤,严重者致多器官功能衰竭,死亡率高.本文观察血液稀释(HD)对致死性鲩鱼胆汁中毒的治疗作用.实验采用 SD 大鼠53只.随机分为中毒组(n=25),稀释治疗组(n=21),正常对照组(n=7).实验结果表明,稀释治疗组动物的谷丙转氨酶(SGPT),黄疽指数(Ⅱ).尿素氮(BUN),升高幅度低于中毒组,肺、肝、肾病理组织学损伤轻于中毒组,24小时内死亡率中毒组高于治疗组,提示 HD 对致死性鲩鱼胆汁中毒可能有保护作用.     

Progress in crystallization of major histocompatibility complex class I in vertebrates

The major histocompatibility complex(MHC)of proteins that exists in all vertebrates is encoded by a cluster of genes associated with the immune response and related functions.MHC is divided into MHC I,II,and III;MHC I is involved in antigenic presentation,binding T cell receptors,and leading ultimately to specific cellular immune responses.The complicated functions of MHC I are determined by the nature of the complex.The crystal structure of MHC I has been solved for many animals,revealing the relationship between spatial structure and function.MHC I consists of an a heavy chain and a b2m light chain,both ligated non-covalently to a complex when a peptide is bound to the antigenic-binding groove.The a heavy chain is divided into an extracellular domain,a transmembrane domain,and an intracellular domain.The extracellular domain consists of sub-regions a1,a2,and a3.The a1 and a2 together form the antigenic-binding groove and bind antigenic peptides with 8–10 amino acid residues.MHC I can form a stable spatial structure;however,it should be noted that there are differences in the structure of MHC I among animal species,including anchored amino acids in binding peptides,binding sites,molecular distance,crystallization conditions,etc.Here,progress in determination of the crystal structure of human,mouse,chicken,non-human primate,and swine MHC I is described in detail.