Stable, soluble, high-affinity, engineered T cell receptors: novel antibody-like proteins for specific targeting of peptide antigens

Boulter, Jonathan M.; Jakobsen, Bent K.

doi:10.1111/j.1365-2249.2005.02929.x

Cited by 25 publications

(16 citation statements)

References 64 publications

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“…In vitro engineering of TCRs for increased affinity has been accomplished using two different methods (yeast and phage display), but so far with limited success (Boulter and Jakobsen, 2005; Richman and Kranz, 2007). The major obstacle to success has been the difficulties associated with the expression of the αβ heterodimer in heterologous display systems, a feature that has for many decades plagued efforts to express soluble TCRs (Rudolph et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…the lower the affinity, the greater the number of ligands required) (Schodin et al, 1996). One method to overcome this issue has been to engineer higher affinity TCRs in vitro (Boulter and Jakobsen, 2005; Richman and Kranz, 2007), with the hope of introducing the novel α and β chain genes into T cells for adoptive transfer into patients, in order to fight the malignancy (i.e. in vitro TCR gene therapy followed by adoptive T cell therapy) (Schumacher, 2002; Blattman and Greenberg, 2004; Engels and Uckert, 2007; Rosenberg et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…Two TCRs, A6 and 1G4 (both MHC class-I restricted), have been engineered for higher affinity using this method (Li et al, 2005). Phage display of these TCRs was enabled by introduction of a non-native disulfide bond between the two C domains in order to promote pairing of the α and β chains (Boulter and Jakobsen, 2005). An advantage of this format is that it allowed for the engineering of the full-length (VαCα/VβCβ) protein without the need for prior selection of V domain stabilizing mutations.…”

Section: Introductionmentioning

confidence: 99%

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Engineering higher affinity T cell receptors using a T cell display system

Chervin

Aggen

Raseman

et al. 2008

Journal of Immunological Methods

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The T cell receptor (TCR) determines the cellular response to antigens, which are presented on the surface of target cells in the form of a peptide bound to a product of the major histocompatibility complex (pepMHC). The response of the T cell depends on the affinity of the TCR for the pepMHC, yet many TCRs have been shown to be of low affinity, and some naturally occurring T cell responses are poor due to low affinities. Accordingly, engineering the TCR for increased affinity for pepMHC, particularly tumor-associated antigens, has become an increasingly desirable goal, especially with the advent of adoptive T cell therapies. For largely technical reasons, to date there have been only a handful of TCRs engineered in vitro for higher affinity using well established methods of protein engineering. Here we report the use of a T cell display system, using a retroviral vector, for generating a high affinity TCR from the mouse T cell clone 2C. The method relies on the display of the TCR, in its normal, signaling competent state, as a CD3 complex on the T cell surface. A library in the CDR3α of the 2C TCR was generated in the MSCV retroviral vector and transduced into a TCRnegative hybridoma. Selection of a high affinity, CD8-independent TCR was accomplished after only two rounds of flow cytometric sorting using the pepMHC SIYRYYGL/K b (SIY/K b ). The selected TCR contained a sequence motif in the CDR3α with characteristics of several other TCRs previously selected by yeast display. In addition, it was possible to directly use the selected T cell hybridoma in functional assays without the need for sub-cloning, revealing that the selected TCR was capable of mediating CD8-independent activity. The method may be useful in the direct isolation and characterization of TCRs that could be used in therapies with adoptive transferred T cells.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Engineering higher affinity T cell receptors using a T cell display system

Chervin

Aggen

Raseman

et al. 2008

Journal of Immunological Methods

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“…Most typically, TCR α and β chain genes are inserted into retroviral vectors and used to transduce activated host T lymphocytes (Engels and Uckert, 2007; Udyavar et al, 2009). Other approaches include lentiviral transduction (Circosta et al, 2009; Zhou et al, 2003), transposon-mediated gene integration (Peng et al, 2009), and direct gene transfer (Cooper et al, 2006). …”

Section: Tcr-modified T Lymphocytes For Tumor Immunotherapymentioning

confidence: 99%

“…Several approaches have been used that can enhance TCR pairing. Cysteine substitutions can be introduced in the constant regions of the α and β chains that lead to the formation of a new interchain disulfide bond (Boulter and Jakobsen, 2005; Cohen et al, 2006; Kuball et al, 2007). Other approaches include directly linking CD3ζ to TCR chains (Sebestyen et al, 2008), use of single chain TCRs (Lake et al, 1999; Zhang et al, 2004), mutating complementary Cα and Cβ interacting residues (Voss et al, 2008), and substituting human Cα and Cβ with their murine counterparts (Cohen et al, 2006; Voss et al, 2006).…”

Section: Optimizing Tcr Expressionmentioning

confidence: 99%

Rebalancing Immune Specificity and Function in Cancer by T-Cell Receptor Gene Therapy

Udyavar

Geiger

2010

Arch. Immunol. Ther. Exp.

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Adoptive immunotherapy with tumor-specific T lymphocytes has demonstrated clinical benefit in some cancers, particularly melanoma. Yet isolating and expanding tumor-specific cells from patients is challenging, and there is limited ability to control T cell affinity and response characteristics. T cell receptor (TCR) gene therapy, in which T lymphocytes for immunotherapy are redirected using introduced rearranged TCR, has emerged as an important alternative. Successful TCR gene therapy requires consideration of a number of issues, including TCR specificity and affinity, optimal gene therapy constructs, types of T cells administered, and the survival and activity of the modified cells. In this review, we highlight the rationale for and experience with, as well as new approaches to enhance TCR gene therapy.

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