H-2 Antigens pp 177-184 | Cite as

Site Directed Mutagenesis Identifies Allo-Antigenic Epitopes of an H-2 Antigen Recognized by Antibodies and by Cytotoxic T-Lymphocytes

  • Keiko Ozato
  • David Koeller
  • Ronald Lieberman
  • Jun-ichi Miyazaki
  • Ettore Appella
  • Donald W. Mann
  • James Forman
Part of the NATO ASI Series book series (NSSA, volume 144)

Abstract

To test our previous hypothesis that the segment between amino acid position 63 to 73 of the H-2Dd antigen forms a major allo-antigenic site, mutations were introduced into the H-2Ld gene in a sequential fashion, which replaced the codons for amino acid position 63, 65, 66, 70 and 73 of the H-2Ld antigen with those of the H-2Dd antigen. Gain and loss of serological and CTL epitopes specific for the H-2Dd and H-2Ld antigens were examined. The amino acid substitutions at position 63, 65, and 66 led to the acquisition of multiple serological H-2Dd specificities which were expressed in the mutant H-2Ld antigen. A further amino acid substitution at position 70 resulted in the gain of additional H-2Dd specificities, allowing to localize more than half of all the relevant H-2Dd serological epitopes to position 63 to 70. An alto CTL epitope of the H-2Dd antigen was also localized to this stretch of amino acid sequence, as one of several H-2Dd specific CTL clones reacted with the mutant molecule in which amino acids were replaced at position 63 to 70. Further, some H-2Ld specific alto CTL clones lost reactivity to the mutant molecules, demonstrating the presence of CTL epitopes in this region of the H-2Ld antigen. From these results we conclude that the amino acid sequence encompassing from position 63 to 70 of the H-2Dd and H-2Ld molecules forms major alto- antigenic epitopes recognized by multiple antibodies and CTLs.

Keywords

Major Histocompatibility Complex Class Amino Acid Position External Domain Amphipathic Structure Mutant Molecule 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1987

Authors and Affiliations

  • Keiko Ozato
    • 1
  • David Koeller
    • 1
  • Ronald Lieberman
    • 1
    • 3
  • Jun-ichi Miyazaki
    • 1
  • Ettore Appella
    • 2
  • Donald W. Mann
    • 4
  • James Forman
    • 4
  1. 1.Laboratory of Developmental and Molecular ImmunityNational Institute of Child Health and Human DevelopmentUSA
  2. 2.Laboratory of Cell BiologyNational Cancer Institute, National Institutes of HealthBethesdaUSA
  3. 3.Division of Cardio-Reno Drug and Drug ProductsFood and Drug AdministrationRockvilleUSA
  4. 4.Department of MicrobiologyUniversity of Texas, Health Science Center at DallasDallasUSA

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