γδTCR immunoglobulin constant region domain exchange in human αβTCRs improves TCR pairing without altering TCR gene-modified T cell function

Tao,Changli; Shao,Hongwei; Zhang,Wenfeng; Bo,Huaben; Wu,Fenglin; Shen,Han; Huang,Shulin

doi:10.3892/mmr.2017.6206

γδTCR immunoglobulin constant region domain exchange in human αβTCRs improves TCR pairing without altering TCR gene-modified T cell function

Authors:
- Changli Tao
- Hongwei Shao
- Wenfeng Zhang
- Huaben Bo
- Fenglin Wu
- Han Shen
- Shulin Huang
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Affiliations: Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, P.R. China
Published online on: February 15, 2017 https://doi.org/10.3892/mmr.2017.6206
Pages: 1555-1564
Copyright: © Tao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The adoptive genetic transfer of T cell receptors (TCRs) has been shown to be overall feasible and offer clinical potential as a treatment for different types of cancer. However, this promising clinical approach is limited by the serious potential consequence that exogenous TCR mispairing with endogenous TCR chains may lead to the risk of self-reactivity. In the present study, domain‑exchange and three‑dimensional modeling strategies were used to create a set of chimeric TCR variants, which were used to exchange the partial or complete constant region of αβTCR with corresponding γδTCR domains. The expression, assembly and function of the chimeric TCR variants were examined in Jurkat T cells and peripheral mononuclear blood cells (PBMCs). Genetically‑encoded chimeras were fused with a pair of fluorescent proteins (ECFP/EYFP) to monitor expression and the pairing between chimeric TCRα chains and TCRβ chains. The fluorescence energy transfer based on confocal laser scanning microscopy showed that the introduction of γδTCR constant sequences into the αβTCR did not result in a global reduction of mispairing with endogenous TCR. However, the TCR harboring the immunoglobulin‑like domain of the γδTCR constant region (i.e., TCR∆IgC), showed a higher expression and preferential pairing, compared with wild‑type (wt)TCR. The function analysis showed that TCR∆IgC exhibited the same levels of interferon-γ production and cytotoxic activity, compared with wtTCR. Furthermore, these modified TCR-transduced T cells retained the classic human leukocyte antigen restriction of the original TCR. The other two chimeric TCRs, had either exchange of the cp+tm+ic domain or exchange of the whole C domain (Fig. 1). Ultimately, exchange of these domains demonstrated defective function in the transduced T cells. Taken together, these findings may provide further understanding of the γδTCR constant domain with implications for the improvement of TCR gene transfer therapy.

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1	Sharpe M and Mount N: Genetically modified T cells in cancer therapy: Opportunities and challenges. Dis Model Mech. 8:337–350. 2015. View Article : Google Scholar : PubMed/NCBI
2	Casucci M, Hawkins RE, Dotti G and Bondanza A: Overcoming the toxicity hurdles of genetically targeted T cells. Cancer Immunol Immunother. 64:123–130. 2015. View Article : Google Scholar : PubMed/NCBI
3	Cameron BJ, Gerry AB, Dukes J, Harper JV, Kannan V, Bianchi FC, Grand F, Brewer JE, Gupta M, Plesa G, et al: Identification of a Titin-derived HLA-A1-presented peptide as a cross-reactive target for engineered MAGE A3-directed T cells. Sci Transl Med. 5:197ra1032013. View Article : Google Scholar : PubMed/NCBI
4	Johnson LA, Morgan RA, Dudley ME, Cassard L, Yang JC, Hughes MS, Kammula US, Royal RE, Sherry RM, Wunderlich JR, et al: Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood. 114:535–546. 2009. View Article : Google Scholar : PubMed/NCBI
5	Parkhurst MR, Yang JC, Langan RC, Dudley ME, Nathan DA, Feldman SA, Davis JL, Morgan RA, Merino MJ, Sherry RM, et al: T cells targeting carcinoembryonic antigen can mediate regression of metastatic colorectal cancer but induce severe transient colitis. Mol Ther. 19:620–626. 2011. View Article : Google Scholar
6	Morgan RA, Chinnasamy N, Abate-Daga D, Gros A, Robbins PF, Zheng Z, Dudley ME, Feldman SA, Yang JC, Sherry RM, et al: Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy. J Immunother. 36:133–151. 2013. View Article : Google Scholar : PubMed/NCBI
7	Linette GP, Stadtmauer EA, Maus MV, Rapoport AP, Levine BL, Emery L, Litzky L, Bagg A, Carreno BM, Cimino PJ, et al: Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma. Blood. 122:863–871. 2013. View Article : Google Scholar : PubMed/NCBI
8	van Loenen MM, De Boer R, Amir AL, Hagedoorn RS, Volbeda GL, Willemze R, van Rood JJ, Falkenburg JH and Heemskerk MH: Mixed T cell receptor dimers harbor potentially harmful neoreactivity. Proc Natl Acad Sci USA. 107:10972–10977. 2010. View Article : Google Scholar : PubMed/NCBI
9	Bendle GM, Linnemann C, Hooijkaas AI, Bies L, De Witte MA, Jorritsma A, Kaiser AD, Pouw N, Debets R, Kieback E, et al: Lethal graft-versus-host disease in mouse models of T cell receptor gene therapy. Nat Med. 16:565–570. 2010. View Article : Google Scholar : PubMed/NCBI
10	Cohen CJ, Zhao Y, Zheng Z, Rosenberg SA and Morgan RA: Enhanced antitumor activity of murine-human hybrid T-cell receptor (TCR) in human lymphocytes is associated with improved pairing and TCR/CD3 stability. Cancer Res. 66:8878–8886. 2006. View Article : Google Scholar : PubMed/NCBI
11	Cohen CJ, Li YF, El-Gamil M, Robbins PF, Rosenberg SA and Morgan RA: Enhanced antitumor activity of T cells engineered to express T-cell receptors with a second disulfide bond. Cancer Res. 67:3898–3903. 2007. View Article : Google Scholar : PubMed/NCBI
12	Voss RH, Willemsen RA, Kuball J, Grabowski M, Engel R, Intan RS, Guillaume P, Romero P, Huber C and Theobald M: Molecular design of the Calphabeta interface favors specific pairing of introduced TCRalphabeta in human T cells. J Immunol. 180:391–401. 2008. View Article : Google Scholar : PubMed/NCBI
13	Aggen DH, Chervin AS, Schmitt TM, Engels B, Stone JD, Richman SA, Piepenbrink KH, Baker BM, Greenberg PD, Schreiber H and Kranz DM: Single-chain VαVβ T-cell receptors function without mispairing with endogenous TCR chains. Gene Ther. 19:365–374. 2012. View Article : Google Scholar : PubMed/NCBI
14	Ochi T, Fujiwara H, Okamoto S, An J, Nagai K, Shirakata T, Mineno J, Kuzushima K, Shiku H and Yasukawa M: Novel adoptive T-cell immunotherapy using a WT1-specific TCR vector encoding silencers for endogenous TCRs shows marked antileukemia reactivity and safety. Blood. 118:1495–1503. 2011. View Article : Google Scholar : PubMed/NCBI
15	Provasi E, Genovese P, Lombardo A, Magnani Z, Liu PQ, Reik A, Chu V, Paschon DE, Zhang L, Kuball J, et al: Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer. Nat Med. 18:807–815. 2012. View Article : Google Scholar : PubMed/NCBI
16	van der Veken LT, Coccoris M, Swart E, Falkenburg JH, Schumacher TN and Heemskerk MH: Alpha beta T cell receptor transfer to gamma delta T cells generates functional effector cells without mixed TCR dimers in vivo. J Immunol. 182:164–170. 2009. View Article : Google Scholar : PubMed/NCBI
17	Marcu-Malina V, Heijhuurs S, van Buuren M, Hartkamp L, Strand S, Sebestyen Z, Scholten K, Martens A and Kuball J: Redirecting αβ T cells against cancer cells by transfer of a broadly tumor-reactive γδT-cell receptor. Blood. 118:50–59. 2011. View Article : Google Scholar : PubMed/NCBI
18	Tao C, Shao H, Yuan Y, Wang H, Zhang W, Zheng W, Ma W and Huang S: Imaging of T-cell receptor fused to CD3ζ reveals enhanced expression and improved pairing in living cells. Int J Mol Med. 34:849–855. 2014.PubMed/NCBI
19	Jang YY, Cho D, Kim SK, Shin DJ, Park MH, Lee JJ, Shin MG, Shin JH, Suh SP and Ryang DW: An improved flow cytometry-based natural killer cytotoxicity assay involving calcein AM staining of effector cells. Ann Clin Lab Sci. 42:42–49. 2012.PubMed/NCBI
20	De Libero G, Lau SY and Mori L: Phosphoantigen presentation to TCR γδ Cells, a conundrum getting less gray zones. Front immunol. 5:6792014.PubMed/NCBI
21	Scheper W, Sebestyen Z and Kuball J: Cancer Immunotherapy using γδT cells: Dealing with diversity. Front Immunol. 5:6012014. View Article : Google Scholar : PubMed/NCBI
22	Kuhns MS, Davis MM and Garcia KC: Deconstructing the form and function of the TCR/CD3 complex. Immunity. 24:133–139. 2006. View Article : Google Scholar : PubMed/NCBI
23	Heemskerk MH, Hagedoorn RS, van der Hoorn MA, van der Veken LT, Hoogeboom M, Kester MG, Willemze R and Falkenburg JH: Efficiency of T-cell receptor expression in dual-specific T cells is controlled by the intrinsic qualities of the TCR chains within the TCR-CD3 complex. Blood. 109:235–243. 2007. View Article : Google Scholar : PubMed/NCBI
24	Teixeiro E, Daniels MA, Hausmann B, Schrum AG, Naeher D, Luescher I, Thome M, Bragado R and Palmer E: T cell division and death are segregated by mutation of TCRbeta chain constant domains. Immunity. 21:515–526. 2004. View Article : Google Scholar : PubMed/NCBI