Distinct epitopes are recognized by cytolytic T lymphocyte clones on the same class I molecule: Direct demonstration using DNA-transfected targets and long-term cytolytic T cell clones

E. McLaughlin-Taylor, J. G. Woodward, M. McMillan, Jeffrey A Frelinger

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Abstract

By producing long-term, stable, cytolytic T lymphocyte clones and utilizing targets expressing only a single gene product derived from the stimulator mouse strain, we have been able to directly demonstrate that T cells recognize distinct epitopes expressed on a single H-2 molecule. These multiple determinants are distinguishable by inhibition patterns with monoclonal antibodies (mAb). When two T cell clones, P-2.14 and P-2.17, are tested on a L cell transfected with the D(P) gene (λ12a), the T cells kill the transfected targets as well as blasts derived from D(P) mouse strains. mAb 7-16.10 inhibits recognition and killing of D(P) targets by both P-2.14 and P-2.17. This mAb recognizes a private specificity H-2.m22. Interestingly mAb 11-20.3 which also recognizes the H-2.m22 specificity inhibits clone P-2.14 but not P-2.17. The mAb 7-16.10, however, competes with 11-20.3 for binding to the surface of L cells expressing the D(P) gene. Thus the two T cells must recognize an overlapping specificity. Other mAb which bind to the H-2D(P) molecule are unable to inhibit either of these two cytolytic T cell clones. Paradoxically, any of the mAb when tested individually are sufficient to inhibit the polyclonal response derived from in vitro mixed lymphocyte culture. Therefore, by using targets expressing only a single H-2 molecule derived by DNA-mediated gene transfer and cytolytic T cell clones we have been able to directly demonstrate the presence of multiple epitopes on a single molecule and define their inhibition with mAb directed to that same molecule.

Original languageEnglish (US)
Pages (from-to)969-974
Number of pages6
JournalEuropean Journal of Immunology
Volume14
Issue number11
StatePublished - 1984
Externally publishedYes

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Epitopes
Clone Cells
Monoclonal Antibodies
T-Lymphocytes
DNA
Genes
Lymphocytes

ASJC Scopus subject areas

  • Immunology

Cite this

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title = "Distinct epitopes are recognized by cytolytic T lymphocyte clones on the same class I molecule: Direct demonstration using DNA-transfected targets and long-term cytolytic T cell clones",
abstract = "By producing long-term, stable, cytolytic T lymphocyte clones and utilizing targets expressing only a single gene product derived from the stimulator mouse strain, we have been able to directly demonstrate that T cells recognize distinct epitopes expressed on a single H-2 molecule. These multiple determinants are distinguishable by inhibition patterns with monoclonal antibodies (mAb). When two T cell clones, P-2.14 and P-2.17, are tested on a L cell transfected with the D(P) gene (λ12a), the T cells kill the transfected targets as well as blasts derived from D(P) mouse strains. mAb 7-16.10 inhibits recognition and killing of D(P) targets by both P-2.14 and P-2.17. This mAb recognizes a private specificity H-2.m22. Interestingly mAb 11-20.3 which also recognizes the H-2.m22 specificity inhibits clone P-2.14 but not P-2.17. The mAb 7-16.10, however, competes with 11-20.3 for binding to the surface of L cells expressing the D(P) gene. Thus the two T cells must recognize an overlapping specificity. Other mAb which bind to the H-2D(P) molecule are unable to inhibit either of these two cytolytic T cell clones. Paradoxically, any of the mAb when tested individually are sufficient to inhibit the polyclonal response derived from in vitro mixed lymphocyte culture. Therefore, by using targets expressing only a single H-2 molecule derived by DNA-mediated gene transfer and cytolytic T cell clones we have been able to directly demonstrate the presence of multiple epitopes on a single molecule and define their inhibition with mAb directed to that same molecule.",
author = "E. McLaughlin-Taylor and Woodward, {J. G.} and M. McMillan and Frelinger, {Jeffrey A}",
year = "1984",
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T1 - Distinct epitopes are recognized by cytolytic T lymphocyte clones on the same class I molecule

T2 - Direct demonstration using DNA-transfected targets and long-term cytolytic T cell clones

AU - McLaughlin-Taylor, E.

AU - Woodward, J. G.

AU - McMillan, M.

AU - Frelinger, Jeffrey A

PY - 1984

Y1 - 1984

N2 - By producing long-term, stable, cytolytic T lymphocyte clones and utilizing targets expressing only a single gene product derived from the stimulator mouse strain, we have been able to directly demonstrate that T cells recognize distinct epitopes expressed on a single H-2 molecule. These multiple determinants are distinguishable by inhibition patterns with monoclonal antibodies (mAb). When two T cell clones, P-2.14 and P-2.17, are tested on a L cell transfected with the D(P) gene (λ12a), the T cells kill the transfected targets as well as blasts derived from D(P) mouse strains. mAb 7-16.10 inhibits recognition and killing of D(P) targets by both P-2.14 and P-2.17. This mAb recognizes a private specificity H-2.m22. Interestingly mAb 11-20.3 which also recognizes the H-2.m22 specificity inhibits clone P-2.14 but not P-2.17. The mAb 7-16.10, however, competes with 11-20.3 for binding to the surface of L cells expressing the D(P) gene. Thus the two T cells must recognize an overlapping specificity. Other mAb which bind to the H-2D(P) molecule are unable to inhibit either of these two cytolytic T cell clones. Paradoxically, any of the mAb when tested individually are sufficient to inhibit the polyclonal response derived from in vitro mixed lymphocyte culture. Therefore, by using targets expressing only a single H-2 molecule derived by DNA-mediated gene transfer and cytolytic T cell clones we have been able to directly demonstrate the presence of multiple epitopes on a single molecule and define their inhibition with mAb directed to that same molecule.

AB - By producing long-term, stable, cytolytic T lymphocyte clones and utilizing targets expressing only a single gene product derived from the stimulator mouse strain, we have been able to directly demonstrate that T cells recognize distinct epitopes expressed on a single H-2 molecule. These multiple determinants are distinguishable by inhibition patterns with monoclonal antibodies (mAb). When two T cell clones, P-2.14 and P-2.17, are tested on a L cell transfected with the D(P) gene (λ12a), the T cells kill the transfected targets as well as blasts derived from D(P) mouse strains. mAb 7-16.10 inhibits recognition and killing of D(P) targets by both P-2.14 and P-2.17. This mAb recognizes a private specificity H-2.m22. Interestingly mAb 11-20.3 which also recognizes the H-2.m22 specificity inhibits clone P-2.14 but not P-2.17. The mAb 7-16.10, however, competes with 11-20.3 for binding to the surface of L cells expressing the D(P) gene. Thus the two T cells must recognize an overlapping specificity. Other mAb which bind to the H-2D(P) molecule are unable to inhibit either of these two cytolytic T cell clones. Paradoxically, any of the mAb when tested individually are sufficient to inhibit the polyclonal response derived from in vitro mixed lymphocyte culture. Therefore, by using targets expressing only a single H-2 molecule derived by DNA-mediated gene transfer and cytolytic T cell clones we have been able to directly demonstrate the presence of multiple epitopes on a single molecule and define their inhibition with mAb directed to that same molecule.

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