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Refinement of HLA gene map** with induced B-cell line mutants

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Abstract

The lymphoma cell line BJAB.B95.8.6 was gamma-irradiated to induce mutations of major histocompatibility complex (MHC) encoded genes. Cloned “wild-type” cells were phenotyped HLA-A1, A2, B 13, 1335, Bw4, Bw6, Cw4, DR5, DRw52, DQwl, DQw3, DPw2, DPw4, GLO1*1, PGM3*2-1, and ME1*0 and possessed two apparently normal chromosome 6s prior to mutagenesis. Loss mutants were selected 5 days after 3 Gy gamma-irradiation employing three complement-fixing monoclonal antibodies specific for HLA-A2 (TÜ101) and Bw4 (TÜ48, TÜ109). Fifteen independently arising mutants were isolated and cloned. Ty** with monospecific alloantisera and cell-mediated lympholysis revealed the presence of HLA-A1, 835, Bw6, Cw4, DR5. DRw52, DQw3, and DPw4 specificities on all mutant clones. HLA-A2, B13, and Bw4 were absent. Mutants differed in their expression of class 11 antigens. One group retained DQw1 and DPw2, another was DQw1−, DPw2+, and a third was DQw1−, DPw2−. Karyoty** of the “wild-type” line and selected mutant clones showed that the loss of HLA specificities correlated with deletions which map the HLA-A and -B loci directly to the distal part of the 6p2l.33 region and the class II genes to the region 6p21.33 (proximal) to 6p21.31 (distal) on the short arm of chromosome 6.

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Abbreviations

CML:

cell-mediated lympholysis

CTX:

cytotoxicity

DBBA:

direct bacterial binding assay

EBV:

Epstein-Barr virus

GLO:

glyoxalase

IBBA:

indirect bacterial binding assay

LU:

lytic units

ME1:

cytoplasmic malic enzyme

MHC:

major histocompatibility complex

MOAB:

monoclonal antibody

NADP:

nicotinamide-adenine dinucleotide phosphate

PGM3:

phosphoglucomutase isozyme 3

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Authors and Affiliations

  1. Immunology Laboratory, Medizinische Klinik, Abt. II, Universität Tübingen, D-7400, Tübingen, Germany

    B. Springs, C. Müller, G. Pawelec, P. Wernet & A. Ziegler

  2. Medizinische Hochschule Lübeck, Institut für Humangenetik, D-2400, Lübeck, Germany

    C. Fonatsch

  3. Institut für Anthropologie und Humangenetik, Universität Tübingen, D-7400, Tübingen, Germany

    J. Kömpf

Authors
  1. B. Springs
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  2. C. Fonatsch
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  3. C. Müller
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  4. G. Pawelec
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  5. J. Kömpf
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  6. P. Wernet
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  7. A. Ziegler
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In partial fulfillment of Ph.D. thesis requirements.

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Springs, B., Fonatsch, C., Müller, C. et al. Refinement of HLA gene map** with induced B-cell line mutants. Immunogenetics 21, 277–291 (1985). https://doi.org/10.1007/BF00375380

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