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通过表达分析分离编码HLA - DRβ链的不同cDNA克隆。

Isolation of distinct cDNA clones encoding HLA-DR beta chains by use of an expression assay.

作者信息

Long E O, Wake C T, Strubin M, Gross N, Accolla R S, Carrel S, Mach B

出版信息

Proc Natl Acad Sci U S A. 1982 Dec;79(23):7465-9. doi: 10.1073/pnas.79.23.7465.

DOI:10.1073/pnas.79.23.7465
PMID:6818545
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC347360/
Abstract

cDNA clones encoding different human Ia antigen beta chains were isolated by use of a complementation-expression assay in Xenopus oocytes. The assay was based on two previous findings. First, oocytes injected with mRNA from a human B-cell line express HLA-DR antigen. The three intracellular DR chains are assembled in oocytes and can be immunoprecipitated with anti-DR monoclonal antibodies. Second, we have isolated cDNA clones encoding DR alpha and intermediate chains. In order to identify beta-chain cDNA clones, mRNA was hybrid-selected with pools of cDNA clones, mixed with mRNA for the alpha and intermediate chains, and injected into oocytes. We isolated two distinct clones that could select DR beta-chain mRNA as demonstrated by assembly of the translation product with DR alpha chains and immunoprecipitation with DR-specific monoclonal antibodies. One clone is specific for a beta chain of the DR locus. The other clone, much weaker in its ability to select DR mRNA, encodes another Ia-like beta chain. Full-length cDNA clones corresponding to the DR and Ia-like beta chains were isolated and compared. Cross-hybridization was detectable in the coding regions but not in the 3' untranslated regions. Distinct RNAs homologous to the DR and the Ia-like beta-chain clones were present in B cells but were undetectable in three T-cell lines.

摘要

通过在非洲爪蟾卵母细胞中进行互补表达分析，分离出了编码不同人类Ia抗原β链的cDNA克隆。该分析基于之前的两项发现。第一，注射来自人类B细胞系mRNA的卵母细胞表达HLA - DR抗原。三条细胞内DR链在卵母细胞中组装，并能用抗DR单克隆抗体进行免疫沉淀。第二，我们已经分离出了编码DRα链和中间链的cDNA克隆。为了鉴定β链cDNA克隆，将mRNA与cDNA克隆池进行杂交选择，与α链和中间链的mRNA混合，然后注射到卵母细胞中。我们分离出了两个不同的克隆，如翻译产物与DRα链组装以及用DR特异性单克隆抗体进行免疫沉淀所证明的，它们能够选择DRβ链mRNA。一个克隆对DR基因座的β链具有特异性。另一个克隆选择DR mRNA的能力较弱，编码另一种Ia样β链。分离出了与DR和Ia样β链相对应的全长cDNA克隆并进行了比较。在编码区可检测到交叉杂交，但在3'非翻译区未检测到。与DR和Ia样β链克隆同源的不同RNA存在于B细胞中，但在三个T细胞系中未检测到。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/770afcc029fa/pnas00462-0394-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/e7661c23fab6/pnas00462-0392-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/bb5d7abea792/pnas00462-0393-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/fd5a1f566b36/pnas00462-0394-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/770afcc029fa/pnas00462-0394-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/e7661c23fab6/pnas00462-0392-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/bb5d7abea792/pnas00462-0393-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/fd5a1f566b36/pnas00462-0394-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/347360/770afcc029fa/pnas00462-0394-b.jpg

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本文引用的文献

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Genetics of HLA disease association.HLA疾病关联的遗传学
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The complete sequence of the mRNA for the HLA-DR-associated invariant chain reveals a polypeptide with an unusual transmembrane polarity.与HLA - DR相关的恒定链的mRNA完整序列揭示了一种具有异常跨膜极性的多肽。
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Structural polymorphism of the DC1 light chains from DR1-, DR2-, and DRw6-positive cell lines.来自DR1、DR2和DRw6阳性细胞系的DC1轻链的结构多态性。
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Isolation of cDNA clones for the p33 invariant chain associated with HLA-DR antigens.与HLA-DR抗原相关的p33恒定链cDNA克隆的分离
Proc Natl Acad Sci U S A. 1983 Sep;80(18):5714-8. doi: 10.1073/pnas.80.18.5714.
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Human B cell variants immunoselected against a single Ia antigen subset have lost expression of several Ia antigen subsets.针对单个Ia抗原亚群进行免疫选择的人B细胞变体已失去了几个Ia抗原亚群的表达。
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Gamma-interferon induces expression of the HLA-D antigens on normal and leukemic human myeloid cells.γ干扰素可诱导人正常及白血病髓样细胞上HLA - D抗原的表达。
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Complete amino acid sequence of an HLA-DR antigen-like beta chain as predicted from the nucleotide sequence: similarities with immunoglobulins and HLA-A, -B, and -C antigens.根据核苷酸序列预测的一种HLA - DR抗原样β链的完整氨基酸序列：与免疫球蛋白以及HLA - A、- B和 - C抗原的相似性。
Proc Natl Acad Sci U S A. 1982 Jun;79(12):3687-91. doi: 10.1073/pnas.79.12.3687.
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Isolation and identification of a cDNA clone corresponding to an HLA-DR antigen beta chain.一个与HLA - DR抗原β链对应的cDNA克隆的分离与鉴定。
Proc Natl Acad Sci U S A. 1982 Mar;79(6):1703-7. doi: 10.1073/pnas.79.6.1703.
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[Primary structure of class II human histocompatibility antigens. 1st communication. Amino acid sequence of the N-terminal 198 residues of the beta chain of a HLA-Dw2,2;DR2,2-alloantigen (author's transl)].[人类Ⅱ类组织相容性抗原的一级结构。首次通讯。一种HLA-Dw2,2;DR2,2同种异体抗原β链N端198个残基的氨基酸序列（作者译）]
Hoppe Seylers Z Physiol Chem. 1981 Dec;362(12):1665-9.
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Distinct forms of both alpha and beta subunits are present in the human Ia molecular pool.人类Ia分子库中存在α和β亚基的不同形式。
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Characterization of HLA-D-region antigens by two-dimensional gel electrophoresis. Molecular-genotyping.通过二维凝胶电泳对HLA-D区抗原进行表征。分子基因分型。
J Exp Med. 1980 Aug 1;152(2 Pt 2):18s-36s.