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Molecular analysis of an HLA-DP mutant cell line selected for its resistance to killing by HLA-DPw2-specific T-cell clones

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Abstract

A collection of HLA-DP mutants was generated, using ICR 191 as the mutagenic agent and resistance to lysis mediated by HLA-DPw2 allospecific cytotoxic T lymphocytes (CTLs) as the selection criterion. These mutants were derived from the HLA haploid lymphoblastoid cell line 45.1. Loss of HLA-DPw2 surface expression accounted for the lack of HLA-DPw2 CTL recognition in all the mutants. However, one of them, 45.EM19, binds to DPw2-specific monoclonal antibodies (mAb) after cell permeabilization. HLA-DPA1 and DPB1 mRNA expression studies permitted the classification of the mutants in four categories: 1) DPA1-negative, DPB1-positive; 2) DPA1-positive, DPB1-negative; 3) DPA1- and DPB1-negative, and 4) DPA1- and DPB1-positive mutants. Mutant 45.EM19 is included in the last group. The cloning and sequencing of the full-length DPA1 (DPA1*0103) and DPB1 (DPB1*02012) cDNAs from this mutant showed no changes in the DPA1 sequence compared to the wild-type sequence. However, a frame-shift mutation in the DPB1 gene exon coding for the transmembrane region was detected. The insertion of a guanine nucleotide provokes an extension of the open reading frame, increasing the length of the C-terminal domain and changing the hydropathicity pattern of the transmembrane domain. This change should be responsible for the phenotype of the 45.EM19 mutant.

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Authors and Affiliations

  1. Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense, 28040, Madrid, Spain

    Javier Arroyo, Rosalía Díez-Orejas, Alberto M. Álvarez &  Miguel Sánchez-Pérez

  2. Centro de Citometría de Flujo, Universidad Complutense, 28040, Madrid, Spain

    Alberto M. Álvarez &  Miguel Sánchez-Pérez

  3. Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, 20892, Bethesda, MD, USA

    Stephen Shaw

Authors
  1. Javier Arroyo
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  2. Rosalía Díez-Orejas
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  3. Alberto M. Álvarez
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  4. Stephen Shaw
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  5. Miguel Sánchez-Pérez
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Correspondence to: Dr. M. Sánchez-Pérez.

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Arroyo, J., Díez-Orejas, R., Álvarez, A.M. et al. Molecular analysis of an HLA-DP mutant cell line selected for its resistance to killing by HLA-DPw2-specific T-cell clones. Immunogenetics 39, 40–47 (1994). https://doi.org/10.1007/BF00171795

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  • DOI: https://doi.org/10.1007/BF00171795

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