Structural polymorphism of human DR antigens

Silver, Jack; Ferrone, Soldano

doi:10.1038/279436a0

Cited by 63 publications

(23 citation statements)

References 14 publications

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“…Weaker bands visible in the EcoRI and HindII digests most likely represent fragments of the same gene rather than other related genes because (i) the major bands are only 3.2 and 2.4 kilobases (kb), respectively, (ii) washing was at high stringency, and (iii) there was only one BamHI fragment. In addition, no evidence for polymorphism was obtained because the same pattern was observed with three human cell lines of different DR specificities, which is consistent with the biochemical evidence that the a chain is not the polymorphic constituent of HLA-DR (6)(7)(8). …”

Section: Methodssupporting

confidence: 71%

“…After addition of EDTA to 10 mM and Tris HCl (pH 7.6) to 100 mM, the sample was extracted and passed over a Sepharose CL-6B column, and the excluded peak was precipitated. ds cDNA was elongated with dCMP residues in 200 mM potassium cacodylate/50 mM Tris HCl, pH 6.9/1 mM CoCl2/1 mM dCTP containing 100 ,ug ofbovine serum albumin (Pentex, Kankakee, IL; Miles), =2 jig of ds cDNA, and 125 units of terminal deoxynucleotidyltransferase (P-L Biochemicals) per ml; incubation was for [1][2][3][4][5][6] (Fig. 1, lane 1).…”

Section: Methodsmentioning

confidence: 99%

“…Multiple alleles have been described for the HLA-DR antigens. This polymorphism is carried by the 3 chains whereas the a chains are constant in individuals with different DR specificities (6)(7)(8). In addition, recent work with anti-DR monoclonal antibodies has identified at least two distinct molecular subsets or isotypes of DR antigens in homozygous cell lines, implying the existence of multiple DR genes (9).…”

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confidence: 99%

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Isolation of cDNA clones encoding HLA-DR alpha chains.

Wake¹,

Long²,

Strubin³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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HLA-DR antigens, the human equivalent of mouse Ia antigens, are multimeric surface glycoproteins characterized by a high degree of allelic polymorphism. They are expressed specifically on macrophages and lymphocytes and they play a key role in the regulation of the immune response. We have investigated this complex genetic system by a direct study of the genes involved through molecular cloning. This paper deals with the cloning, in plasmids, of full-length cDNA sequences for the HLA-DR a chain from the human B-cell line Raji. The approach relies on a translation assay of mRNA injected into frog oocytes and recognition of translation products by polyclonal and monoclonal antibodies. After enrichment of specific mRNA and cloning ofcDNA, plasmid clones were analyzed by hybridization-selection of mRNA and translation in oocytes. A clone was identified and used to screen a cDNA library from which several full-length HLA-DR a chain plasmids were isolated. DNA sequence determination of one such clone confirmed its identity and also established the amino acid sequence of the NH2-terminal signal sequence ofHLA-DR a chains. The translation product ofHLA-DR a chain mRNA purified by hybridization-selection gives a single a chain spot on two-dimensional gels, whereas the a chain released from the a//3 HLA-DR complex gives about seven distinct spots. Finally, the results ofanalysis ofgenomic DNA by Southern blotting are compatible with the existence of a single nonpolymorphic a chain gene and indicate extensive cross-hybridization with a homologous gene in mouse DNA.

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Section: Methodssupporting

confidence: 71%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Isolation of cDNA clones encoding HLA-DR alpha chains.

Wake¹,

Long²,

Strubin³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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“…Most of the genetic variation in HLA-II molecules is located to the more distal parts of the ectodomains. A highly peculiar exception here is the monomorphic a-chain of HLA-DR, with only one functional allele (39). The inability of HKB1 to bind all HLA-DR molecules made us exclude the a-chain from the analysis, and rather focus on the b-chain.…”

Section: Specificity Of Vaccine Proteins For Human Hla-ii Moleculesmentioning

confidence: 99%

Antigen Targeting to Human HLA Class II Molecules Increases Efficacy of DNA Vaccination

Grødeland

Fredriksen²,

Løset

et al. 2016

The Journal of Immunology

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It has been difficult to translate promising results from DNA vaccination in mice to larger animals and humans. Previously, DNA vaccines encoding proteins that target Ag to MHC class II (MHC-II) molecules on APCs have been shown to induce rapid, enhanced, and long-lasting Ag-specific Ab titers in mice. In this study, we describe two novel DNA vaccines that as proteins target HLA class II (HLA-II) molecules. These vaccine proteins cross-react with MHC-II molecules in several species of larger mammals. When tested in ferrets and pigs, a single DNA delivery with low doses of the HLA-II–targeted vaccines resulted in rapid and increased Ab responses. Importantly, painless intradermal jet delivery of DNA was as effective as delivery by needle injection followed by electroporation. As an indication that the vaccines could also be useful for human application, HLA-II–targeted vaccine proteins were found to increase human CD4+ T cell responses by a factor of ×103 in vitro. Thus, targeting of Ag to MHC-II molecules may represent an attractive strategy for increasing efficacy of DNA vaccines in larger animals and humans.

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“…Since most of the polymorphism attributed to DR molecules has been correlated with structural heterogeneity of the DR (&chain (9)(10)(11)(12)(13), we have prepared cell lines from donors of HLA-D homozygous typing cells representing each of the distinct HLA-D antigens associated with DR4 and have studied the relationships among isolated DR4 ,3-chain polypeptides. Our results demonstrate a striking correlation between the functionally distinct HLA-D phenotypes of individual HCL and the two-dimensional gel patterns of the DR ,B chains isolated from these HCL.…”

mentioning

confidence: 99%

Electrophoretic analysis of human HLA-DR antigens from HLA-DR4 homozygous cell lines: correlation between beta-chain diversity and HLA-D.

Nepom¹,

Nepom²,

Mickelson³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

119

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Two-dimensional gel electrophoresis of immunoprecipitated human HLA-DR antigens from cells expressing the HLA-DR4 haplotype shows distinct clustering of (3-chain patterns. Six unique electrophoretic variants were observed among 17 HLA-DR4 homozygous cell lines (HCL) analyzed. These patterns correlate precisely with the HLA-D phenotype of the HCL donor as determined by reactivity in mixed lymphocyte culture. All DR4 HCL that belong to one of the well-defined HLA-D antigen groups (Dw4, DwlO, LD "40", LD "DYI", LD "KT2", or LD "TAS") have identical DR (-chain . Comprehensive populations studies with DR4-associated HTC have led to the description of several distinct DR4-associated HLA-D antigens, including Dw4, DwlO, LD "40", LD "DYT", and LD "KT2" (4-8). There are several possible explanations for these apparent DR4 "splits." First, HLA-Dw4 and DR4 may be the products of different loci, each coding for distinct "D region"-associated antigens. A second possibility is that Dw4 and DR4 are products of a single gene, the primary product of which is recognized by DR4 alloantisera, but through post-translational modification sufficient phenotype variation occurs to cause the differences detected in MLC by T cells. In a third model, the haplotypes expressing Dw4, DwlO, LD 40, LD DYT, and LD KT2 may be encoded by different alleles but share a common crossreactive or "supertypic" determinant recognized by DR4 alloantisera.To answer such questions, we have begun a detailed analysis of the two-dimensional gel electrophoresis patterns of DR antigens expressed by B-lymphoblastoid cell lines (HCL) derived from HLA-DR4 homozygous donors. Since most of the polymorphism attributed to DR molecules has been correlated with structural heterogeneity of the DR (&chain (9-13), we have prepared cell lines from donors of HLA-D homozygous typing cells representing each of the distinct HLA-D antigens associated with DR4 and have studied the relationships among isolated DR4 ,3-chain polypeptides. Our results demonstrate a striking correlation between the functionally distinct HLA-D phenotypes of individual HCL and the two-dimensional gel patterns of the DR ,B chains isolated from these HCL.MATERIALS AND METHODS HCL. Seventeen B-lymphoblastoid cell lines homozygous for an HLA-DR4-associated D locus were used in this study (Table 1). The specificities Dw4, Dw10, LD 40, LD DYT, LD KT2, and DB3 have been defined by the Eighth Workshop (5), by Reinsmoen and Bach (4), and by Nose et al. (2). In addition, we have used HCL derived from a unique HTC, "TAS," described by Amar et al. (20), that is mutually reactive in MLC with cells from each of the other clusters and that may represent yet another apparent split of the DR4 haplotype.Established B-lymphoblastoid cell lines were available for some of the above HTC, otherwise new B-lymphoblastoid lines were prepared from cryopreserved HTC by cocultivation with Epstein-Barr virus (EBV) as described (21). Briefly, 1-2 X 106

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Structural polymorphism of human DR antigens

Cited by 63 publications

References 14 publications

Isolation of cDNA clones encoding HLA-DR alpha chains.

Isolation of cDNA clones encoding HLA-DR alpha chains.

Antigen Targeting to Human HLA Class II Molecules Increases Efficacy of DNA Vaccination

Electrophoretic analysis of human HLA-DR antigens from HLA-DR4 homozygous cell lines: correlation between beta-chain diversity and HLA-D.

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