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Serological expression after sequential double transfection with purified HLA-11 gene of mouse fibroblasts carrying human beta-2 microglobulin

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Abstract

A genomic cosmid library constructed from DNA from a genotyped individual (JF = HLA-A11, Cw−, B38/A26, Cw7, B51) was screened for clones containing class I histocompatibility genes. Among these clones, one was found to carry a 4.8 kb Hind III fragment which is highly correlated with HLA-A11. This clone was used to transfect LMTK+ cultured mouse fibroblast transformants expressing human beta-2 microglobulin. The human beta-2 microglobulin heavy chain-associated determinant was positively detected by the M18 monoclonal antibody. HLA-A11 expression on these doubly transformed cells was specifically demonstrated by complement-dependent cytotoxicity with HLA-A11 + A3-specific but not with HLA-A3-specific monoclonal antibodies. Absorption studies with human alloantisera confirmed the presence on these cells of HLA-A11 determinants and of cross-reacting determinants which absorbed anti-HLA-A1 and −A3 alloantisera. The JF5-J27 transfected cell expressed both heavy and light chains of human class I histocompatibility genes.

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Abbreviations

β2m:

beta-2 microglobulin

CTL:

cytolytic T lymphocytes

FCS:

fetal calf serum

HAT:

hypoxanthine-azaguanine-thymidine

kb:

kilobase pair

MHC:

major histocompatibility complex

MoAb:

monoclonal antibodies

PBL:

peripheral blood lymphocytes

PEG:

polyethylene glycol

r :

correlation coefficient

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Authors and Affiliations

  1. INSERM U 93, Hôpital St. Louis, 2, place du Dr. Fournier, 75475, Paris Cedex 10, France

    Pascale Paul, Virginia Lepage, Brigitte Sayagh, Marika Pla, Laurence Boumsell, Corinne Douay, Daniel Cohen, Jacques Colombani, Jean Dausset &  Laurent Degos

  2. INSERM U 152, Hôpital Cochin, 27, rue du Fg. St. Jacques, 75013, Paris, France

    Jean-Jacques Metzger

  3. Centre d'Etude du Polymorphisme Humain, Collège de france, place Marcelin Berthelot, 75005, Paris, France

    Jean Dausset

Authors
  1. Pascale Paul
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  2. Virginia Lepage
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  3. Brigitte Sayagh
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  4. Jean-Jacques Metzger
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  5. Marika Pla
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  6. Laurence Boumsell
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  7. Corinne Douay
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  8. Daniel Cohen
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  9. Jacques Colombani
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  10. Jean Dausset
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  11. Laurent Degos
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This study is dedicated to the memory of Jean-Jacques Metzger.

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Paul, P., Lepage, V., Sayagh, B. et al. Serological expression after sequential double transfection with purified HLA-11 gene of mouse fibroblasts carrying human beta-2 microglobulin. Immunogenetics 22, 1–8 (1985). https://doi.org/10.1007/BF00430589

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  • DOI: https://doi.org/10.1007/BF00430589

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