Secretion of a Chimeric T-cell Receptor-immunoglobulin Protein

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1987 May 1

PMID 3472243

Citations 16

Authors

N R Gascoigne

C C Goodnow

K I Dudzik

V T Oi

M M Davis

Affiliations

Soon will be listed here.

Abstract

To produce sufficient quantities of soluble T-cell receptor protein for detailed biochemical and biophysical analyses we have explored the use of immunoglobulin--T-cell receptor gene fusions. In this report we describe a chimeric gene construct containing a T-cell receptor alpha-chain variable (V) domain and the constant (C) region coding sequences of an immunoglobulin gamma 2a molecule. Cells transfected with the chimeric gene synthesize a stable protein product that expresses immunoglobulin and T-cell receptor antigenic determinants as well as protein A binding sites. We show that the determinant recognized by the anticlonotypic antibody A2B4.2 resides on the V alpha domain of the T-cell receptor. The chimeric protein associates with a normal lambda light chain to form an apparently normal tetrameric (H2L2, where H = heavy and L = light) immunoglobulin molecule that is secreted. Also of potential significance is the fact that a T-cell receptor V beta gene in the same construct is neither assembled nor secreted with the lambda light chain, and when expressed with a C kappa region it does not assemble with the chimeric V alpha C gamma 2a protein mentioned above. This indicates that not all T-cell receptor V regions are similar enough to immunoglobulin V regions for them to be completely interchangeable.

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References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Kindred B, Shreffler D . H-2 dependence of co-operation between T and B cells in vivo. J Immunol. 1972; 109(5):940-3. View

Katz D, Hamaoka T, Benacerraf B . Cell interactions between histoincompatible T and B lymphocytes. II. Failure of physiologic cooperative interactions between T and B lymphocytes from allogeneic donor strains in humoral response to hapten-protein conjugates. J Exp Med. 1973; 137(6):1405-18. PMC: 2139346. DOI: 10.1084/jem.137.6.1405. View

Shulman M, Wilde C, Kohler G . A better cell line for making hybridomas secreting specific antibodies. Nature. 1978; 276(5685):269-70. DOI: 10.1038/276269a0. View

Zinkernagel R, Doherty P . MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Adv Immunol. 1979; 27:51-177. DOI: 10.1016/s0065-2776(08)60262-x. View

Deisenhofer J . Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from Staphylococcus aureus at 2.9- and 2.8-A resolution. Biochemistry. 1981; 20(9):2361-70. View

Mulligan R, Berg P . Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc Natl Acad Sci U S A. 1981; 78(4):2072-6. PMC: 319285. DOI: 10.1073/pnas.78.4.2072. View

Matzinger P, Zamoyska R . A beginner's guide to major histocompatibility complex function. Nature. 1982; 297(5868):628. DOI: 10.1038/297628a0. View

Oi V, Morrison S, Herzenberg L, Berg P . Immunoglobulin gene expression in transformed lymphoid cells. Proc Natl Acad Sci U S A. 1983; 80(3):825-9. PMC: 393473. DOI: 10.1073/pnas.80.3.825. View

10.

Gillies S, Morrison S, Oi V, Tonegawa S . A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983; 33(3):717-28. DOI: 10.1016/0092-8674(83)90014-4. View

11.

Haas I, Wabl M . Immunoglobulin heavy chain binding protein. Nature. 1983; 306(5941):387-9. DOI: 10.1038/306387a0. View

12.

Samelson L, Germain R, Schwartz R . Monoclonal antibodies against the antigen receptor on a cloned T-cell hybrid. Proc Natl Acad Sci U S A. 1983; 80(22):6972-6. PMC: 390108. DOI: 10.1073/pnas.80.22.6972. View

13.

Oi V, Vuong T, Hardy R, Reidler J, Dangle J, Herzenberg L . Correlation between segmental flexibility and effector function of antibodies. Nature. 1984; 307(5947):136-40. DOI: 10.1038/307136a0. View

14.

Yanagi Y, Yoshikai Y, Leggett K, Clark S, Aleksander I, Mak T . A human T cell-specific cDNA clone encodes a protein having extensive homology to immunoglobulin chains. Nature. 1984; 308(5955):145-9. DOI: 10.1038/308145a0. View

15.

Hedrick S, Nielsen E, Kavaler J, Cohen D, Davis M . Sequence relationships between putative T-cell receptor polypeptides and immunoglobulins. Nature. 1984; 308(5955):153-8. DOI: 10.1038/308153a0. View

16.

Siu G, Clark S, Yoshikai Y, Malissen M, Yanagi Y, Strauss E . The human T cell antigen receptor is encoded by variable, diversity, and joining gene segments that rearrange to generate a complete V gene. Cell. 1984; 37(2):393-401. DOI: 10.1016/0092-8674(84)90369-6. View

17.

Chien Y, Gascoigne N, Kavaler J, Lee N, Davis M . Somatic recombination in a murine T-cell receptor gene. Nature. 1984; 309(5966):322-6. DOI: 10.1038/309322a0. View

18.

Sharon J, Gefter M, Manser T, Morrison S, Oi V, Ptashne M . Expression of a VHC kappa chimaeric protein in mouse myeloma cells. Nature. 1984; 309(5966):364-7. DOI: 10.1038/309364a0. View

19.

Malissen M, Minard K, Mjolsness S, Kronenberg M, Goverman J, Hunkapiller T . Mouse T cell antigen receptor: structure and organization of constant and joining gene segments encoding the beta polypeptide. Cell. 1984; 37(3):1101-10. DOI: 10.1016/0092-8674(84)90444-6. View

20.

Gascoigne N, Chien Y, Becker D, Kavaler J, Davis M . Genomic organization and sequence of T-cell receptor beta-chain constant- and joining-region genes. Nature. 1984; 310(5976):387-91. DOI: 10.1038/310387a0. View