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  • 1
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A newly defined leukocyte antigen system, the secondary B-cell (SB) system, was shown to be linked to HLA. The SB marker has been investigated in lymphocyte donors presumed to be genetically homozygous for HLA-A through HLA-D/DR by virtue of descent from a first-cousin marriage and of phenotypic homozygosity for these HLA markers. Of 19 donors, 3 were found to be heterozygous for SB. Studies of the families of these three donors could not distinguish with certainty whether the heterozygosity resulted from SB/DR recombination or from “pseudohomozygosity” for HLA-A through -D/DR by inheritance of two genetically unrelated but similar haplotypes. However, our data favored the occurrence of SB/DR recombination with a meiotic distance perhaps as large as 3.3 cMorgan. Recombinations were identified which mapped SB between HLA-B and GL01. These studies demonstrate the usefulness of cells from first-cousin marriage offspring in mapping a polymorphic genetic system.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract HLA-DR molecules were isolated from eight different HLA-DR3 homozygous B-cell lines by immunoprecipitation with monoclonal antibodies, and they were subsequently analyzed by two-dimensional gel electrophoresis. We found that HLA-DR3 homozygous B-cell lines of consanguineous origin express two types of HLA-DR molecules. One type of HLA-DR molecule was present in all the cell lines tested, whereas the second DR molecule appears to be polymorphic. DNA isolated from the different HLA-DR3 homozygous cell lines was studied by Southern blot analysis to determine whether any DRβ restriction fragment length polymorphism could be observed. Polymorphisms detected at both the product and genomic level have been compared to each other, and their relations to the serological (HLA-DR) and cellular (HLA-D and LB-Q1) typing data will be discussed.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract HLA-DR molecules were isolated from HLA-DR3, −5, and −w6 positive homozygous B-cell lines by immunoprecipitation with monoclonal antibodies and analyzed by gel electrophoretic techniques. DNA isolated from the same cell lines was digested with the restriction enzyme Taq I and hybridized with a DR beta full-length cDNA probe. We demonstrated that certain DR βI alleles are found in combination with different DR βIII alleles as defined by Southern blotting, protein chemistry, a functional assay using purified protein derivative-specific T-cell lines, and, in one case, also alloreactive T-cell reagents. Our results indicate that within the family of HLA-DRw52-associated haplotypes DR beta chain genes may have been transferred from one haplotype to another. The implications of these findings are discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have analyzed the HLA class II gene products from HLA-DRw6 homozygous cells. Epstein-Barr virus-transformed B-cell lines were internally labeled with [35S]-methionine. An NP-40 lysate of the cells was subjected to immunoprecipitation, first with a DRw52-like-specific monoclonal antibody and subsequently with a DR-specific framework antibody. The DR region-encoded gene products were analyzed by one-dimensional gel isoelectric focusing and two-dimensional gel electrophoresis. It is shown that DRw6 homozygous cell lines contain at least two nonallelic DR β chains, one carrying a DRw52 determinant and one DRw52-negative population. Both chains appear to be polymorphic between the cellularly defined subtypes of DRw6. The determinant responsible for the differential mixed lymphocyte culture reactivity of Dw18 and Dw19 cells resides on the DRw52-positive population, whereas the Dw6-Dw9 differences are attributed to determinants on both populations of DR light chains. The Dw16-derived DRw52+ chain much resembles the Dw18 DRw52+ light chain whereas there is a clear-cut difference between these two subtypes in the DRw52− population. We conclude that, for DRw6 homozygous cells, the cellularly recognized D determinants are probably located on DR-encoded molecules, both DRw52+ and DRw52−, and that charge shift of these chains is at least partly responsible for differential recognition of these cells in mixed lymphocyte cultures.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Individuals of known HLA phenotype were used to examine the specificity of secondary proliferative (restimulation) responses in lymphocyte populations which had been primed previously in long-term, one-way mixed lymphocyte reactions (MLR). The objective was to identify the genetic region responsible for this secondary mitogenic effect. Restimulation responses between unrelated and related individuals from a family with a recombinant betweenHLA-B andD produced similar results. In both cases responses were grouped into zero, intermediate, and positive. Restimulation was by three types of cell: those with HLA-D types compatible with the responder; those with HLA-D types specific to the primary stimulator; and those with HLA-D types of a third-party cell. Compatible responses fell into the zero category and the intermediate category, third-party responses fell into the intermediate category, and specific responses fell into the positive category. Our conclusion was that HLA-D products evoked secondary mitogenic responses, while structures coded for by the HLA-A and B regions appeared to have no effect.
    Type of Medium: Electronic Resource
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