Cloning and Expression of Human Membrane-Bound and Soluble Engineered T Cell Receptors for Immunotherapy

Alajez, Nehad M.; Eghtesad, Saman; Finn, Olivera J.

doi:https://doi.org/10.1155/JBB/2006/68091

BioMed Research International

On this page

Abstract References Copyright Related Articles

Methods Report | Open Access

Volume 2006 | Article ID 068091 | https://doi.org/10.1155/JBB/2006/68091

Cloning and Expression of Human Membrane-Bound and Soluble Engineered T Cell Receptors for Immunotherapy

Nehad M. Alajez,¹Saman Eghtesad,¹and Olivera J. Finn¹

Received28 Jun 2005

Revised15 Nov 2005

Accepted21 Nov 2005

Published23 Feb 2006

Abstract

We report here the design and construction of several gene vectors for expression in mammalian cells of membrane-bound and soluble human T cell receptors (TR). We designed a vector (TR-ALPHA-IRES-TR-BETA pEF4) that encodes high-level expression of the full-length TR on the surface of T cells. Furthermore, we engineered TR that does not require the presence of endogenous CD3 molecules for surface expression and thus expression is not limited to T cells. We also constructed a vector encoding a single-chain TR (scTR) as a fusion protein of V-ALPHA-V-BETA-C-BETA with CD3Z. Since it is encoded and expressed as a single molecule, this scTR is well suited for gene therapy. Lastly, we successfully used a mammalian expression vector for generation of soluble human TR. The approaches we used here for manipulation of a human tumor-specific TR can be useful for other investigators interested in TR-based immunotherapy.

References

M E Dudley, J R Wunderlich, P F Robbins et al., “Cancer regression and autoimmunity in patients after clonal repopulation with antitumor lymphocytes,” Science, vol. 298, no. 5594, pp. 850–854, 2002.
View at: Google Scholar
M P Rubinstein, A N Kadima, M L Salem et al., “Transfer of TCR genes into mature T cells is accompanied by the maintenance of parental T cell avidity,” The Journal of Immunology, vol. 170, no. 3, pp. 1209–1217, 2003.
View at: Google Scholar
R A Morgan, M E Dudley, Y Y Yu et al., “High efficiency TCR gene transfer into primary human lymphocytes affords avid recognition of melanoma tumor antigen glycoprotein 100 and does not alter the recognition of autologous melanoma antigens,” The Journal of Immunology, vol. 171, no. 6, pp. 3287–3295, 2003.
View at: Google Scholar
C A Aarnoudse, M Krüse, R Konopitzky, N Brouwenstijn, and P I Schrier, “TCR reconstitution in Jurkat reporter cells facilitates the identification of novel tumor antigens by cDNA expression cloning,” International Journal of Cancer, vol. 99, no. 1, pp. 7–13, 2002.
View at: Google Scholar
M A Derby, J Wang, D H Margulies, and J A Berzofsky, “Two intermediate-avidity cytotoxic T lymphocyte clones with a disparity between functional avidity and MHC tetramer staining,” International Immunology, vol. 13, no. 6, pp. 817–824, 2001.
View at: Google Scholar
J T Snyder, M A Alexander-Miller, J A Berzofsky, and I M Belyakov, “Molecular mechanisms and biological significance of CTL avidity,” Current HIV Research, vol. 1, no. 3, pp. 287–294, 2003.
View at: Google Scholar
M E Weijtens, E H Hart, and R L Bolhuis, “Functional balance between T cell chimeric receptor density and tumor associated antigen density: CTL mediated cytolysis and lymphokine production,” Gene Therapy, vol. 7, no. 1, pp. 35–42, 2000.
View at: Google Scholar
S Yang, G P Linette, S Longerich, and F G Haluska, “Antimelanoma activity of CTL generated from peripheral blood mononuclear cells after stimulation with autologous dendritic cells pulsed with melanoma gp100 peptide G209-2M is correlated to TCR avidity,” The Journal of Immunology, vol. 169, no. 1, pp. 531–539, 2002.
View at: Google Scholar
J Magarian-Blander, P Ciborowski, S Hsia, S C Watkins, and O J Finn, “Intercellular and intracellular events following the MHC-unrestricted TCR recognition of a tumor-specific peptide epitope on the epithelial antigen MUC1,” The Journal of Immunology, vol. 160, no. 7, pp. 3111–3120, 1998.
View at: Google Scholar
I Engel, T H Ottenhoff, and R D Klausner, “High-efficiency expression and solubilization of functional T cell antigen receptor heterodimers,” Science, vol. 256, no. 5061, pp. 1318–1321, 1992.
View at: Google Scholar
M Mittelbrunn, M Yáñez-Mó, D Sancho, A Ursa, and F Sánchez-Madrid, “Cutting edge: dynamic redistribution of tetraspanin CD81 at the central zone of the immune synapse in both T lymphocytes and APC,” The Journal of Immunology, vol. 169, no. 12, pp. 6691–6695, 2002.
View at: Google Scholar
N M Alajez, J Schmielau, M D Alter, M Cascio, and O J Finn, “Therapeutic potential of a tumor-specific, MHC-unrestricted T-cell receptor expressed on effector cells of the innate and the adaptive immune system through bone marrow transduction and immune reconstitution,” Blood, vol. 105, no. 12, pp. 4583–4589, 2005.
View at: Google Scholar
R A Willemsen, M E Weijtens, C Ronteltap et al., “Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR,” Gene Therapy, vol. 7, no. 16, pp. 1369–1377, 2000.
View at: Google Scholar
J Novotny, R K Ganju, S T Smiley et al., “A soluble, single-chain T-cell receptor fragment endowed with antigen-combining properties,” Proceedings of the National Academy of Sciences of the United States of America, vol. 88, no. 19, pp. 8646–8650, 1991.
View at: Google Scholar
Z Eshhar, N Bach, C J Fitzer-Attas et al., “The T-body approach: potential for cancer immunotherapy,” Springer Seminars in Immunopathology, vol. 18, no. 2, pp. 199–209, 1996.
View at: Google Scholar
C Grégoire, S Y Lin, G Mazza, N Rebai, I F Luescher, and B Malissen, “Covalent assembly of a soluble T cell receptor-peptide-major histocompatibility class I complex,” Proceedings of the National Academy of Sciences of the United States of America, vol. 93, no. 14, pp. 7184–7189, 1996.
View at: Google Scholar
G Pavlinkova, D Colcher, B J Booth, A Goel, and S K Batra, “Pharmacokinetics and biodistribution of a light-chain-shuffled CC49 single-chain Fv antibody construct,” Cancer Immunology, Immunotherapy, vol. 49, no. 4-5, pp. 267–275, 2000.
View at: Google Scholar

Copyright

Copyright © 2006 Nehad M. Alajez et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation Order printed copies

Views

207

Downloads

927

Citations