Development of a T-cell Receptor Mimic Antibody against Wild-Type p53 for Cancer Immunotherapy

A particular feature of γδ T cell biology is that cells expressing T cell receptor (TCR) using specific Vγ/Vδ segments are localized in distinct epithelial sites, e.g., in mouse epidermis nearly all γδ T cells express Vγ3/Vδ1. These cells, referred to as dendritic epidermal T cells (DETC) originate from fetal Vγ3+ thymocytes. The role of γδ TCR specificity in DETC's migration/localization to the skin has remained controversial. To address this issue we have generated transgenic (Tg) mice expressing a TCR δ chain (Vδ6.3-Dδ1-Dδ2-Jδ1-Cδ), which can pair with Vγ3 in fetal thymocytes but is not normally expressed by DETC. In wild-type (wt) Vδ6.3Tg mice DETC were present and virtually all of them express Vδ6.3. However, DETC were absent in TCR-δ−/− Vδ6.3Tg mice, despite the fact that Vδ6.3Tg γδ T cells were present in normal numbers in other lymphoid and nonlymphoid tissues. In wt Vδ6.3Tg mice, a high proportion of in-frame Vδ1 transcripts were found in DETC, suggesting that the expression of an endogenous TCR-δ (most probably Vδ1) was required for the development of Vδ6.3+ epidermal γδ T cells. Collectively our data demonstrate that TCR specificity is essential for the development of γδ T cells in the epidermis. Moreover, they show that the TCR-δ locus is not allelically excluded.

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T‐cell receptor affinity in the age of cancer immunotherapy

Molecular Carcinogenesis ◽

10.1002/mc.23212 ◽

2020 ◽

Vol 59 (7) ◽

pp. 862-870 ◽

Cited By ~ 2

Author(s):

Michele M. Hoffmann ◽

Jill E. Slansky

Keyword(s):

T Cell ◽

Cancer Immunotherapy ◽

T Cell Receptor ◽

Cell Receptor ◽

Receptor Affinity

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Influence of the NH2-terminal Amino Acid of the T Cell Receptor α Chain on Major Histocompatibility Complex (MHC) Class II + Peptide Recognition

Journal of Experimental Medicine ◽

10.1084/jem.185.11.1919 ◽

1997 ◽

Vol 185 (11) ◽

pp. 1919-1927 ◽

Cited By ~ 19

Author(s):

Jeffrey L. Seibel ◽

Nancy Wilson ◽

Haruo Kozono ◽

Philippa Marrack ◽

John W. Kappler

Keyword(s):

Major Histocompatibility Complex ◽

Amino Acid ◽

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Wild Type ◽

Terminal Amino Acid ◽

Histocompatibility Complex ◽

Terminal Amino ◽

Α Chain

The α/β T cell receptor (TCR) recognizes peptide fragments bound in the groove of major histocompatibility complex (MHC) molecules. We modified the TCR α chain from a mouse T cell hybridoma and tested its ability to reconstitute TCR expression and function in an α chain–deficient variant of the hybridoma. The modified α chain differed from wild type only in its leader peptide and mature NH2-terminal amino acid. Reconstituted cell surface TCR complexes reacted normally with anti-TCR and anti-CD3 antibodies. Although cross-linking of this TCR with an antibody to the TCR idiotype elicited vigorous T cell hybridoma activation, stimulation with its natural MHC + peptide ligand did not. We demonstrated that this phenotype could be reproduced simply by substituting the glutamic acid (E) at the mature NH2 terminus of the wild type TCR α chain with aspartic acid (D). The substitution also dramatically reduced the affinity of soluble α/β-TCR heterodimers for soluble MHC + peptide molecules in a cell-free system, suggesting that it did not exert its effect simply by disrupting TCR interactions with accessory molecules on the hybridoma. These results demonstrate for the first time that amino acids which are not in the canonical TCR complementarity determining regions can be critical in determining how the TCR engages MHC + peptide.

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GATA3 Abundance Is a Critical Determinant of T Cell Receptor β Allelic Exclusion

Molecular and Cellular Biology ◽

10.1128/mcb.00052-17 ◽

2017 ◽

Vol 37 (12) ◽

Cited By ~ 3

Author(s):

Chia-Jui Ku ◽

JoAnn M. Sekiguchi ◽

Bharat Panwar ◽

Yuanfang Guan ◽

Satoru Takahashi ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

The Other ◽

Allelic Exclusion ◽

Wild Type ◽

Critical Determinant ◽

Gata3 Expression ◽

Immunological Process ◽

Functional Receptor

ABSTRACT Allelic exclusion describes the essential immunological process by which feedback repression of sequential DNA rearrangements ensures that only one autosome expresses a functional T or B cell receptor. In wild-type mammals, approximately 60% of cells have recombined the DNA of one T cell receptor β (TCRβ) V-to-DJ-joined allele in a functional configuration, while the second allele has recombined only the DJ sequences; the other 40% of cells have recombined the V to the DJ segments on both alleles, with only one of the two alleles predicting a functional TCRβ protein. Here we report that the transgenic overexpression of GATA3 leads predominantly to biallelic TCRβ gene (Tcrb) recombination. We also found that wild-type immature thymocytes can be separated into distinct populations based on intracellular GATA3 expression and that GATA3LO cells had almost exclusively recombined only one Tcrb locus (that predicted a functional receptor sequence), while GATA3HI cells had uniformly recombined both Tcrb alleles (one predicting a functional and the other predicting a nonfunctional rearrangement). These data show that GATA3 abundance regulates the recombination propensity at the Tcrb locus and provide new mechanistic insight into the historic immunological conundrum for how Tcrb allelic exclusion is mediated.

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High-throughput determination of the antigen specificities of T cell receptors in single cells

10.1101/457069 ◽

2018 ◽

Cited By ~ 3

Author(s):

Shu-Qi Zhang ◽

Ke-Yue Ma ◽

Alexandra A. Schonnesen ◽

Mingliang Zhang ◽

Chenfeng He ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

High Throughput ◽

Single Cells ◽

Cell Receptor ◽

Cross Reactivity ◽

Wild Type ◽

Type Antigen

We present tetramer-associated T-cell receptor sequencing (TetTCR-Seq), a method to link T cell receptor (TCR) sequences to their cognate antigens in single cells at high throughput. Binding is determined using a library of DNA-barcoded antigen tetramers that is rapidly generated by in vitro transcription and translation. We applied TetTCR-Seq to identify patterns in TCR cross-reactivity with cancer neo-antigens and to rapidly isolate neo-antigen-specific TCRs with no cross-reactivity to the wild-type antigen.

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Promiscuous Behavior of HPV16E6 Specific T Cell Receptor Beta Chains Hampers Functional Expression in TCR Transgenic T Cells, Which Can Be Restored in Part by Genetic Modification

Analytical Cellular Pathology ◽

10.1155/2010/616497 ◽

2010 ◽

Vol 32 (1-2) ◽

pp. 43-56

Author(s):

Kirsten B. J. Scholten ◽

Janneke J. Ruizendaal ◽

Marcus Graf ◽

Thomas Schoedl ◽

Duco Kramer ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Receptor Gene ◽

Careful Analysis ◽

Cell Receptor ◽

Open Reading Frames ◽

Wild Type ◽

Human T Cells ◽

Reading Frames

Background: T cell receptor gene transfer is a promising strategy to treat patients suffering from HPV induced malignancies. Therefore we isolated the TCRαβ open reading frames of an HPV16E6 specific CTL clone and generated TCR transgenic T cells. In general low level expression of the transgenic TCR in recipient human T cells is observed as well as the formation of mixed TCRs dimers. Here we addressed both issues employing three different expression platforms.Methods: We isolated the HVP16E6 specific TCRα and TCRβ open reading frames and retrovirally transduced human T cells with either wild-type (wt), or codon-modified (cm) chains to achieve enhanced TCR expression levels, or used codon-modification in combination with cysteinization (cmCys) of TCRs to facilitate preferential pairing of the introduced TCRα and TCRβ chains.Results: Careful analysis of recipient T cells carrying the HPV16E6 TCRβ and endogenous TCR chains revealed the transgenic TCRβ chain to behave very promiscuously. Further analysis showed that the percentage of tetramer positive T cells in codon-modified/cysteinized TCR transgenic T cells was four-fold higher compared to wild-type and two-fold higher compared to codon-modification only. Functional activity, as determined by IFN-γ production, was high in cmCysTCR transgenic T cells, where it was low in cm and wt TCR transgenic T cells. Recognition of endogenously processed HPV16E6 antigen by cmCysTCR transgenic T cells was confirmed in a cytotoxicity assay.Conclusions: Promiscuous behavior of the HPV16E6 specific TCRβ chain can in part be forced back into specific action in TCR transgenic T cells by codon modification in combination with the inclusion of an extra cysteine in the TCR chains.

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A Robotic Microscope System to Examine T Cell Receptor Acuity Against Tumor Neoantigens: A New Tool for Cancer Immunotherapy Research

IEEE Robotics and Automation Letters ◽

10.1109/lra.2019.2894466 ◽

2019 ◽

Vol 4 (2) ◽

pp. 1760-1767

Author(s):

Lee-Ling Sharon Ong ◽

Hai Zhu ◽

Debasis Banik ◽

Zhenping Guan ◽

Yinnian Feng ◽

...

Keyword(s):

T Cell ◽

Cancer Immunotherapy ◽

T Cell Receptor ◽

Cell Receptor

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Cd8− T Cell Transfectants That Express a High Affinity T Cell Receptor Exhibit Enhanced Peptide-Dependent Activation

Journal of Experimental Medicine ◽

10.1084/jem.194.8.1043 ◽

2001 ◽

Vol 194 (8) ◽

pp. 1043-1052 ◽

Cited By ~ 80

Author(s):

Phillip D. Holler ◽

Alice R. Lim ◽

Bryan K. Cho ◽

Laurie A. Rund ◽

David M. Kranz

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Cytotoxic T Lymphocyte ◽

Cell Activity ◽

Cell Receptor ◽

Wild Type ◽

High Affinity ◽

Antigen Presenting

T cells are activated by binding of the T cell receptor (TCR) to a peptide-major histocompatibility complex (MHC) complex (pMHC) expressed on the surface of antigen presenting cells. Various models have predicted that activation is limited to a narrow window of affinities (or dissociation rates) for the TCR–pMHC interaction and that above or below this window, T cells will fail to undergo activation. However, to date there have not been TCRs with sufficiently high affinities in order to test this hypothesis. In this report we examined the activity of a CD8-negative T cell line transfected with a high affinity mutant TCR (KD = 10 nM) derived from cytotoxic T lymphocyte clone 2C by in vitro engineering. The results show that despite a 300-fold higher affinity and a 45-fold longer off-rate compared with the wild-type TCR, T cells that expressed the mutant TCRs were activated by peptide. In fact, activation could be detected at significantly lower peptide concentrations than with T cells that expressed the wild-type TCR. Furthermore, binding and functional analyses of a panel of peptide variants suggested that pMHC stability could account for apparent discrepancies between TCR affinity and T cell activity observed in several prior studies.

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