Changing T cell specificity by retroviral T cell receptor display

Kessels, Helmut W.; Boom, Marly D. van den; Spits, Hergen; Hooijberg, Erik; Schumacher, Ton N.

doi:10.1073/pnas.97.26.14578

Cited by 87 publications

(56 citation statements)

References 40 publications

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“…But, even if our findings are not universally true for all TCRs, they are of significant importance. With sufficient care, it should be possible to choose or even artificially generate TCRs that display optimized quality (18,87,88).…”

Section: Discussionmentioning

confidence: 99%

Transfer of TCR Genes into Mature T Cells Is Accompanied by the Maintenance of Parental T Cell Avidity

Rubinstein

Kadima

Salem

et al. 2003

The Journal of Immunology

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The adoptive transfer of tumor-specific T cells expanded in vitro can be of significant therapeutic value in select cancer patients. This strategy is limited though, as it is often difficult, if not impossible, to obtain T cells of clinical value. The transfer of TCR genes to mature T cells to generate tumor-reactive T cells provides a potential mechanism to overcome these limitations. To evaluate the feasibility of such an approach and the quality of the resulting T cells, we generated replication-deficient retroviral vectors using the well-characterized OT-1 TCR genes. After transducing murine T cells, we were able to expand large numbers of Ag-specific T cells that were functionally active against tumor cells expressing the relevant Ag. Furthermore, we found that T cells expressing retrovirally encoded TCR had avidity that was similar to that of the parental clone. This maintenance of avidity was despite variable expression of the retrovirally encoded TCR and the presence of potentially competing endogenous TCRs. These results suggest that the inherent qualities of the TCR, as dictated by the coding sequence, are the most critical parameters in the generation of high-avidity T cells.

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Section: Discussionmentioning

confidence: 99%

Transfer of TCR Genes into Mature T Cells Is Accompanied by the Maintenance of Parental T Cell Avidity

Rubinstein

Kadima

Salem

et al. 2003

The Journal of Immunology

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“…The CD28-negative variant Jurkat cell line J.RT3-T3.5 was grown in Iscove's modified Dulbecco's medium (Life Technologies, Paisley, Scotland) with 5% FCS (BioWhittaker). Jurkat cells were transduced with pMX vectors encoding CD28 wt or CD28 KNI by using a transduction protocol as described (17) and sorted for CD28-expressing cells. A total of 2 ϫ 10 7 Jurkat͞CD28 wt or Jurkat͞CD28 KNI cells (in 500 l PBS) was stimulated with 10 g͞ml anti-CD28 mAb 37.51 in the presence of 0.1 mM pervanadate for 20 min on ice.…”

Section: Methodsmentioning

confidence: 99%

Specificity and affinity motifs for Grb2 SH2-ligand interactions

Kessels

Ward

Schumacher

2002

Proc. Natl. Acad. Sci. U.S.A.

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Protein-protein interactions are often mediated by the recognition of short continuous amino acid stretches on target proteins by specific binding domains. Affinity-based selection strategies have successfully been used to define recognition motifs for a large series of such protein domains. However, in many biological systems specificity of interaction may be of equal or greater importance than affinity. To address this issue we have developed a peptide library screening technology that can be used to directly define ligands for protein domains based on both affinity and specificity of interaction. We demonstrate the value of this approach by the selection of peptide ligands that are either highly specific for the Grb2 Src homology 2 (SH2) domain or that are cross-reactive between a group of related SH2 domains. Examination of previously identified physiological ligands for the Grb2 SH2 domain suggests that for these ligands regulation of the specificity of interaction may be an important factor for in vivo ligand selection.T he intracellular organization that enables a cell to respond to external stimuli consists of a complex web of signal transduction pathways. Key factors in the regulation of cellular signaling are the protein binding domains-small, conserved protein modules that mediate intracellular protein-protein interactions. Many of these domains, such as the families of SH2 (Src homology 2), SH3 (Src homology 3), PTB (phosphotyrosine binding), or PDZ (postsynaptic density-95͞Discs large͞zona occludens-1) domains, use short peptide sequences for ligand recognition (1). For example, the binding of SH2 domains to target proteins involves the recognition of a phosphorylated tyrosine residue, and specificity of individual SH2 domains is mediated by the recognition of amino acid residues immediately C-terminal to the phospho-tyrosine (2). The binding preferences of SH2 domains have been studied extensively through the use of peptide libraries, and predictions for the optimal binding motifs for a large number of SH2 domains have been obtained in this manner (3,4). In addition, such binding motifs have been used extensively as lead structures for the design of selective small-molecular SH2 inhibitors (5, 6). Importantly, in traditional library screening strategies used to define SH2 ligand motifs the selection of ligands is exclusively based on the strength of the SH2-phospho-peptide interaction. Consequently, the motifs that are identified in this manner describe ligands with an optimal affinity for a given SH2 domain (here named affinity motifs). Because both affinity and specificity of protein interactions are controlled by the same thermodynamic factors (shape and charge complementarity in the ground state), the selection of high affinity ligands will often also result in the selection of highly specific ligands. However, it has previously been argued that for closely related targets (such as the families of SH2 domains and other signal transduction modules) affinity-based selections may result in the ide...

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“…It has been shown that TCR reconstruction on TCR-deficient T cells is advantageous to investigate how the TCR-pMHC interaction influences T cell activation (27)(28)(29)(30) because primary T cells can increase or decrease their sensitivity/avidity for an epitope through changes in their inhibitory receptor expression and membrane organization as well as via a redistribution of signaling molecules in ϩ cells isolated from an HIV-negative donor were pulsed with various concentrations of VY8 or RY11 peptide (A), infected with recombinant vaccinia virus expressing Nef SF2 (B), or infected with HIV-1 (C) and then mixed with the indicated CTL clones. To obtain relative specific lysis values, the cytotoxic activity toward the same target cells not pulsed with peptide, infected with vaccinia virus alone (i.e., lacking nef expression) or infected with HIV-1 ⌬nef variant was determined in parallel and was deducted as a background value.…”

Section: Functional Reconstruction Of Tcrs On Tcr-deficient T Cellsmentioning

confidence: 99%

Impact of Intrinsic Cooperative Thermodynamics of Peptide-MHC Complexes on Antiviral Activity of HIV-Specific CTL

Motozono

Yanaka

Tsumoto

et al. 2009

The Journal of Immunology

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The antiviral activity of HIV-specific CTL is not equally potent but rather is dependent on their specificity. But what characteristic of targeted peptides influences CTL antiviral activity remains elusive. We addressed this issue based on HLA-B35-restricted CTLs specific for two overlapping immunodominant Nef epitopes, VY8 (VPLRPMTY) and RY11 (RPQVPLRPMTY). VY8-specific CTLs were more potently cytotoxic toward HIV-infected primary CD4+ cells than RY11-specific CTLs. Reconstruction of their TCR revealed no substantial difference in their functional avidity toward cognate Ags. Instead, the decay analysis of the peptide-MHC complex (pMHC) revealed that the VY8/HLA-B35 complex could maintain its capacity to sensitize T cells much longer than its RY11 counterpart. Corroboratively, the introduction of a mutation in the epitopes that substantially delayed pMHC decay rendered Nef-expressing target cells more susceptible to CTL killing. Moreover, by using differential scanning calorimetry and circular dichroism analyses, we found that the susceptible pMHC ligands for CTL killing showed interdependent and cooperative, rather than separate or sequential, transitions within their heterotrimer components under the thermally induced unfolding process. Collectively, our results highlight the significant effects of intrinsic peptide factors that support cooperative thermodynamics within pMHC on the efficient CTL killing of HIV-infected cells, thus providing us better insight into vaccine design.

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Changing T cell specificity by retroviral T cell receptor display

Cited by 87 publications

References 40 publications

Transfer of TCR Genes into Mature T Cells Is Accompanied by the Maintenance of Parental T Cell Avidity

Transfer of TCR Genes into Mature T Cells Is Accompanied by the Maintenance of Parental T Cell Avidity

Specificity and affinity motifs for Grb2 SH2-ligand interactions

Impact of Intrinsic Cooperative Thermodynamics of Peptide-MHC Complexes on Antiviral Activity of HIV-Specific CTL

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