Expression of a hybrid immunoglobulin-T cell receptor protein in transgenic mice

doi:10.1016/0092-8674(89)90943-4

Cell

Volume 58, Issue 5, 8 September 1989, Pages 911-921

https://doi.org/10.1016/0092-8674(89)90943-4 Get rights and content

Abstract

We have constructed a hybrid immunoglobulin (VDJ_H)-T cell receptor (C_α) gene using the VDJ_H exon from a digoxin-specific antibody. This gene was used to make a line of transgenic mice. The hybrid VDJ_H-C_α protein is expressed on a subset of T cells in these mice, and we have shown that it forms part of a functional TCR complex by the criteria of coprecipitation and comodulation of CD3 and TCR β chain components and T cell activation with anti-idiotypic antibodies or digoxin. Furthermore, in cells expressing the hybrid protein, there is allelic exclusion of endogenous TCR α genes. We discuss the implications for the comparative structure of T cell receptors and immunoglobulins.

References (50)

D. Beale et al.
Unusual features of the T-cell receptor C domains are revealed by structural comparisons with other members of the Immunoglobulin superfamily
Comp. Biochem. Physiol.
(1986)
J.W. Kappler et al.
T cell tolerance by clonal elimination in the thymus
Cell
(1987)
Y. Kuwana et al.
Expression of chimeric receptor composed of immunoglobulin-derived V regions and T-cell receptor-derived C regions
Biochem. Biophys. Res. Commun.
(1987)
M. Mudgett-Hunter et al.
Binding and structural diversity among high-affinity monoclonal anti-digoxin antibodies
Mol. Immunol.
(1985)
G. Russo et al.
Molecular analysis of a t(7;14) (q35;q32) chromosome translocation in a T cell leukemia of a patient with Ataxia Telangiectasia
Cell
(1988)
L.E. Samelson et al.
Identification of the components of the murine T cell antigen receptor complex
Cell
(1985)
E. Southern
Detection of specific sequences among DNA fragments separated by agarose gel electrophoresis
J. Mol. Biol.
(1975)
K. Zinn et al.
Identification of two distinct regulatory regions adjacent to the human β-interferon gene
Cell
(1983)
J. Allison et al.
Tumor-specific antigen of murine T-lymphoma defined with monoclonal antibody
J. Immunol.
(1982)
M.J. Bevan
In a radiation chimaera, host H-2 antigens determine immune responsiveness of donor cytotoxic cells
Nature
(1977)

J.A. Bluestone et al.

Structure and specificity of T cell receptor $γ δ$ on major histocompatibility complex antigen-specific CD3⁺, CD4⁻, CD8⁻ T lymphocytes

J. Exp. Med.

(1988)

H. Blüthmann et al.

T-cell-specific expression of T-cell receptor transgenes allows functional rearrangement of endogenous α- and β-genes

Nature

(1988)

L. Bougueleret et al.

Variability analysis of the human and mouse T-cell receptor β chains

Immunogenetics

(1987)

Y.-H. Chien et al.

A new T-cell receptor gene located within the alpha locus and expressed early in T-cell differentiation

Nature

(1987)

C. Chothia et al.

The outline structure of the T-cell αβ receptor

EMBO J.

(1988)

H. Clevers et al.

The T cells receptor/CD3 complex: a dynamic protein ensemble

Annu. Rev. Immunol.

(1988)

I.N. Crispe et al.

Selective activation of Lyt2⁺ precursor T cells by ligation of the antigen receptor

Nature

(1985)

M.M. Davis et al.

T-cell antigen receptor genes and T-cell recognition

Nature

(1988)

Z. Dembic et al.

Transfer of specificity by murine α and β T-cell receptor genes

Nature

(1986)

C.T. Denny et al.

A chromosome 14 inversion in a T-cell lymphoma is caused by site-specific recombination between immunoglobulin and T-cell receptor loci

Nature

(1986)

C. de Preval et al.

Specific interaction between V_H and V_L regions of human monoclonal immunoglobulins

J. Mol. Biol.

(1976)

J.-P. de Villartay et al.

Deletion of the human T-cell receptor δ-gene by a site-specific recombination

Nature

(1988)

N.R.J. Gascoigne et al.

Secretion of a chimeric T-cell receptor-immunoglobulin protein

K. Haskins et al.

The major histocompatibility complex-restricted antigen receptor in T cells. I. Isolation with a monoclonal antibody

J. Exp. Med.

(1983)

S.M. Hedrick et al.

Sequence relationships between putative T-cell receptor polypeptides and immunoglobulins

Nature

(1984)

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Cell

ArticleExpression of a hybrid immunoglobulin-T cell receptor protein in transgenic mice

Abstract

Comp. Biochem. Physiol.

Cell

Biochem. Biophys. Res. Commun.

Mol. Immunol.

Cell

Cell

J. Mol. Biol.

Cell

Tumor-specific antigen of murine T-lymphoma defined with monoclonal antibody

J. Immunol.

In a radiation chimaera, host H-2 antigens determine immune responsiveness of donor cytotoxic cells

Nature

Structure and specificity of T cell receptor γδ on major histocompatibility complex antigen-specific CD3+, CD4−, CD8− T lymphocytes

J. Exp. Med.

T-cell-specific expression of T-cell receptor transgenes allows functional rearrangement of endogenous α- and β-genes

Nature

Variability analysis of the human and mouse T-cell receptor β chains

Immunogenetics

A new T-cell receptor gene located within the alpha locus and expressed early in T-cell differentiation

Nature

The outline structure of the T-cell αβ receptor

EMBO J.

The T cells receptor/CD3 complex: a dynamic protein ensemble

Annu. Rev. Immunol.

Selective activation of Lyt2+ precursor T cells by ligation of the antigen receptor

Nature

T-cell antigen receptor genes and T-cell recognition

Nature

Transfer of specificity by murine α and β T-cell receptor genes

Nature

A chromosome 14 inversion in a T-cell lymphoma is caused by site-specific recombination between immunoglobulin and T-cell receptor loci

Nature

Specific interaction between VH and VL regions of human monoclonal immunoglobulins

J. Mol. Biol.

Deletion of the human T-cell receptor δ-gene by a site-specific recombination

Nature

Secretion of a chimeric T-cell receptor-immunoglobulin protein

The major histocompatibility complex-restricted antigen receptor in T cells. I. Isolation with a monoclonal antibody

J. Exp. Med.

Sequence relationships between putative T-cell receptor polypeptides and immunoglobulins

Nature

Article
Expression of a hybrid immunoglobulin-T cell receptor protein in transgenic mice

Structure and specificity of T cell receptor $γ δ$ on major histocompatibility complex antigen-specific CD3⁺, CD4⁻, CD8⁻ T lymphocytes

Selective activation of Lyt2⁺ precursor T cells by ligation of the antigen receptor

Specific interaction between V_H and V_L regions of human monoclonal immunoglobulins