Abstract 1425: Chimeric antigen receptors based on T cell receptor-like antibodies

AbstractNeoepitopes or neoantigens are a spectrum of unique mutations presented in a particular patient’s tumor. Neoepitope-based adoptive therapies have the potential of tumor eradication without undue damaging effect on normal tissues. In this context, methods based on the T cell receptor (TCR) engineering or chimeric antigen receptors (CARs) have shown great promise. This review focuses on the TCR-like CARs and TCR-CARs directed against tumor-derived epitopes, with a concerted view on neoepitopes. We also address the current limitations of the field to know how to harness the full benefits of this approach and thereby design a sustained and specific antitumor therapy.

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A novel method to generate T-cell receptor–deficient chimeric antigen receptor T cells

Blood Advances ◽

10.1182/bloodadvances.2017012823 ◽

2018 ◽

Vol 2 (5) ◽

pp. 517-528 ◽

Cited By ~ 14

Author(s):

Takahiro Kamiya ◽

Desmond Wong ◽

Yi Tian Png ◽

Dario Campana

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Chimeric Antigen Receptor ◽

Cell Receptor ◽

Surface Expression ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Key Points ◽

Novel Method

Key Points Newly designed PEBLs prevent surface expression of T-cell receptor in T cells without affecting their function. Combined with chimeric antigen receptors, PEBLs can rapidly generate powerful antileukemic T cells without alloreactivity.

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Ig alpha and Ig beta are functionally homologous to the signaling proteins of the T-cell receptor

Molecular and Cellular Biology ◽

10.1128/mcb.14.2.1095-1103.1994 ◽

1994 ◽

Vol 14 (2) ◽

pp. 1095-1103

Author(s):

A L Burkhardt ◽

T Costa ◽

Z Misulovin ◽

B Stealy ◽

J B Bolen ◽

...

Keyword(s):

Signal Transduction ◽

T Cells ◽

T Cell ◽

B Cell ◽

T Cell Receptor ◽

Interleukin 2 ◽

Antigen Receptor ◽

Cell Receptor ◽

Antigen Receptors ◽

Cell Antigen

Signal transduction by antigen receptors and some Fc receptors requires the activation of a family of receptor-associated transmembrane accessory proteins. One common feature of the cytoplasmic domains of these accessory molecules is the presence is at least two YXXA repeats that are potential sites for interaction with Src homology 2 domain-containing proteins. However, the degree of similarity between the different receptor-associated proteins varies from that of T-cell receptor (TCR) zeta and Fc receptor RIIIA gamma chains, which are homologous, to the distantly related Ig alpha and Ig beta proteins of the B-cell antigen receptor. To determine whether T- and B-cell antigen receptors are in fact functionally homologous, we have studied signal transduction by chimeric immunoglobulins bearing the Ig alpha or Ig beta cytoplasmic domain. We found that Ig alpha and Ig beta cytoplasmic domains were able to activate Ca2+ flux, interleukin-2 secretion, and phosphorylation of the same group of cellular substrates as the TCR in transfected T cells. Chimeric proteins were then used to examine the minimal requirements for activation of the Fyn, Lck, and ZAP kinases in T cells. Both Ig alpha and Ig beta were able to trigger Fyn, Lck, and ZAP directly without involvement of TCR components. Cytoplasmic tyrosine residues in Ig beta were required for recruitment and activation of ZAP-70, but these amino acids were not essential for the activation of Fyn and Lck. We conclude that Fyn and Lck are able to recognize a clustered nonphosphorylated immune recognition receptor, but activation of these kinases is not sufficient to induce cellular responses such as Ca2+ flux and interleukin-2 secretion. In addition, the molecular structures involved in antigen receptor signaling pathways are conserved between T and B cells.

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The gamma and epsilon subunits of the CD3 complex inhibit pre-Golgi degradation of newly synthesized T cell antigen receptors.

The Journal of Cell Biology ◽

10.1083/jcb.110.4.973 ◽

1990 ◽

Vol 110 (4) ◽

pp. 973-986 ◽

Cited By ~ 50

Author(s):

T Wileman ◽

G R Carson ◽

M Concino ◽

A Ahmed ◽

C Terhorst

Keyword(s):

T Cells ◽

Endoplasmic Reticulum ◽

Plasma Membrane ◽

T Cell ◽

T Cell Receptor ◽

Intracellular Transport ◽

Cell Receptor ◽

Dna Transfection ◽

Antigen Receptors ◽

Intracellular Degradation

The T cell receptor for antigen (TCR) is composed of six different transmembrane proteins. T cells carefully control the intracellular transport of the receptor and allow only complete receptors to reach the plasma membrane. In an attempt to understand how T cells regulate this process, we used c-DNA transfection and subunit-specific antibodies to follow the intracellular transport of five subunits (alpha beta gamma delta epsilon) of the receptor. In particular, we assessed the intracellular stability of each chain. Our results showed that the chains were markedly different in their susceptibility to intracellular degradation. TCR alpha and beta and CD3 delta were degraded rapidly, whereas CD3 gamma and epsilon were stable. An analysis of the N-linked oligosaccharides of the glycoprotein subunits suggested that the chains were unable to reach the medial Golgi during the metabolic chase. This was supported by immunofluorescence micrographs that showed both the stable CD3 gamma and unstable CD3 delta chain localized in the endoplasmic reticulum. To study the effects of subunit associations on intracellular transport we used cotransfection to reconstitute precise combinations of subunits. Associations between stable and unstable subunits expressed in the same cell led to the formation of stable complexes. These complexes were retained in or close to the endoplasmic reticulum. The results suggested that the intracellular transport of the T cell receptor could be regulated by two mechanisms. The TCR alpha and beta and CD3 delta subunits were degraded rapidly and as a consequence failed to reach the plasma membrane. CD3 gamma or epsilon were stable but were retained inside the cell. The results also demonstrated that there was an interplay between the two pathways such that the CD3 gamma and epsilon subunits were able to protect labile chains from rapid intracellular degradation. In this way, they could seed subunit assembly in or close to the endoplasmic reticulum and allow a stable receptor to form before its transport to the plasma membrane.

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Breast cancer is marked by specific, Public T-cell receptor CDR3 regions shared by mice and humans

PLoS Computational Biology ◽

10.1371/journal.pcbi.1008486 ◽

2021 ◽

Vol 17 (1) ◽

pp. e1008486

Author(s):

Miri Gordin ◽

Hagit Philip ◽

Alona Zilberberg ◽

Moriah Gidoni ◽

Raanan Margalit ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Sequence Data ◽

Breast Tumors ◽

Cell Receptor ◽

Beta Chain ◽

Antigen Receptors ◽

Partial Success

The partial success of tumor immunotherapy induced by checkpoint blockade, which is not antigen-specific, suggests that the immune system of some patients contain antigen receptors able to specifically identify tumor cells. Here we focused on T-cell receptor (TCR) repertoires associated with spontaneous breast cancer. We studied the alpha and beta chain CDR3 domains of TCR repertoires of CD4 T cells using deep sequencing of cell populations in mice and applied the results to published TCR sequence data obtained from human patients. We screened peripheral blood T cells obtained monthly from individual mice spontaneously developing breast tumors by 5 months. We then looked at identical TCR sequences in published human studies; we used TCGA data from tumors and healthy tissues of 1,256 breast cancer resections and from 4 focused studies including sequences from tumors, lymph nodes, blood and healthy tissues, and from single cell dataset of 3 breast cancer subjects. We now report that mice spontaneously developing breast cancer manifest shared, Public CDR3 regions in both their alpha and beta and that a significant number of women with early breast cancer manifest identical CDR3 sequences. These findings suggest that the development of breast cancer is associated, across species, with biomarker, exclusive TCR repertoires.

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Somatic hypermutation of T cell receptor α chain contributes to selection in nurse shark thymus

eLife ◽

10.7554/elife.28477 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 15

Author(s):

Jeannine A Ott ◽

Caitlin D Castro ◽

Thaddeus C Deiss ◽

Yuko Ohta ◽

Martin F Flajnik ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Somatic Hypermutation ◽

Cytidine Deaminase ◽

Cell Receptor ◽

Antigen Receptors ◽

Cell Repertoire ◽

Nurse Shark ◽

Α Chain ◽

Complementarity Determining Regions

Since the discovery of the T cell receptor (TcR), immunologists have assigned somatic hypermutation (SHM) as a mechanism employed solely by B cells to diversify their antigen receptors. Remarkably, we found SHM acting in the thymus on α chain locus of shark TcR. SHM in developing shark T cells likely is catalyzed by activation-induced cytidine deaminase (AID) and results in both point and tandem mutations that accumulate non-conservative amino acid replacements within complementarity-determining regions (CDRs). Mutation frequency at TcRα was as high as that seen at B cell receptor loci (BcR) in sharks and mammals, and the mechanism of SHM shares unique characteristics first detected at shark BcR loci. Additionally, fluorescence in situ hybridization showed the strongest AID expression in thymic corticomedullary junction and medulla. We suggest that TcRα utilizes SHM to broaden diversification of the primary αβ T cell repertoire in sharks, the first reported use in vertebrates.

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T cell receptor repertoire for a viral epitope in humans is diversified by tolerance to a background major histocompatibility complex antigen.

Journal of Experimental Medicine ◽

10.1084/jem.182.6.1703 ◽

1995 ◽

Vol 182 (6) ◽

pp. 1703-1715 ◽

Cited By ~ 138

Author(s):

S R Burrows ◽

S L Silins ◽

D J Moss ◽

R Khanna ◽

I S Misko ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Specific Response ◽

Major Histocompatibility ◽

Antigen Receptors ◽

Barr Virus ◽

Receptor Repertoire ◽

Tcr Repertoire ◽

Epstein Barr

Two unusual characteristics of the memory response to the immunodominant Epstein-Barr virus (EBV) epitope FLRGRAYGL, which associates with HLA B8, have provided an unique opportunity to investigate self tolerance and T cell receptor (TCR) plasticity in humans. First, the response is exceptionally restricted, dominated by cytotoxic T lymphocytes (CTL) with identical TCR protein sequences (Argaet, V. P., C. W. Schmidt, S. R. Burrows, S. L. Silins, M. G. Kurilla, D. L. Doolan, A. Suhrbier, D. J. Moss, E. Kieff, T. B. Sculley, and I. S. Misko. 1994. J. Exp. Med. 180:2335-2340). Second, CTL expressing this receptor are cross-reactive with the alloantigen HLA B* 4402 on uninfected cells (Burrows, S. R., R. Khanna, J. M. Burrows, and D. J. Moss. 1994. J. Exp. Med. 179:1155-1161). No CTL using this conserved public TCR could be reactivated from the peripheral blood of EBV exposed individuals expressing both HLA B8 and B*4402, demonstrating the clonal inactivation of potentially self-reactive T cells in humans. A significant FLRGRAYGL-specific response was still apparent, however, and TCR sequence analysis of multiple CTL clones revealed an oligoclonal TCR repertoire for this determinant within these individuals, using diverse V and J gene segments and CDR3 regions. In addition, a significant public TCR component was identified in which several distinct alpha/beta rearrangements are shared by CTL clones from a number of unrelated HLA B8+, B*4402+ donors. The striking dominance of public TCR in the response to this EBV epitope suggests a strong genetic bias in TCR gene recombination. Fine specificity analysis using peptide analogues showed that, of six different antigen receptors for FLRGRAYGL/HLA B8, none associate closely with the peptide's full array of potential TCR contact residues. Whereas the HLA B*4402-cross-reactive receptor binds amino acids toward the COOH terminus of the peptide, others preferentially favor an NH2-terminal determinant, presumably evading an area that mimics a structure presented on HLA B*4402. Thus, tolerance to a background major histocompatibility antigen can effectively diversify the TCR repertoire for a foreign epitope by deflecting the response away from an immunodominant combination of TCR-binding residues.

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Ig alpha and Ig beta are functionally homologous to the signaling proteins of the T-cell receptor.

Molecular and Cellular Biology ◽

10.1128/mcb.14.2.1095 ◽

1994 ◽

Vol 14 (2) ◽

pp. 1095-1103 ◽

Cited By ~ 41

Author(s):

A L Burkhardt ◽

T Costa ◽

Z Misulovin ◽

B Stealy ◽

J B Bolen ◽

...

Keyword(s):

Signal Transduction ◽

T Cells ◽

T Cell ◽

B Cell ◽

T Cell Receptor ◽

Interleukin 2 ◽

Antigen Receptor ◽

Cell Receptor ◽

Antigen Receptors ◽

Cell Antigen

Signal transduction by antigen receptors and some Fc receptors requires the activation of a family of receptor-associated transmembrane accessory proteins. One common feature of the cytoplasmic domains of these accessory molecules is the presence is at least two YXXA repeats that are potential sites for interaction with Src homology 2 domain-containing proteins. However, the degree of similarity between the different receptor-associated proteins varies from that of T-cell receptor (TCR) zeta and Fc receptor RIIIA gamma chains, which are homologous, to the distantly related Ig alpha and Ig beta proteins of the B-cell antigen receptor. To determine whether T- and B-cell antigen receptors are in fact functionally homologous, we have studied signal transduction by chimeric immunoglobulins bearing the Ig alpha or Ig beta cytoplasmic domain. We found that Ig alpha and Ig beta cytoplasmic domains were able to activate Ca2+ flux, interleukin-2 secretion, and phosphorylation of the same group of cellular substrates as the TCR in transfected T cells. Chimeric proteins were then used to examine the minimal requirements for activation of the Fyn, Lck, and ZAP kinases in T cells. Both Ig alpha and Ig beta were able to trigger Fyn, Lck, and ZAP directly without involvement of TCR components. Cytoplasmic tyrosine residues in Ig beta were required for recruitment and activation of ZAP-70, but these amino acids were not essential for the activation of Fyn and Lck. We conclude that Fyn and Lck are able to recognize a clustered nonphosphorylated immune recognition receptor, but activation of these kinases is not sufficient to induce cellular responses such as Ca2+ flux and interleukin-2 secretion. In addition, the molecular structures involved in antigen receptor signaling pathways are conserved between T and B cells.

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Abnormal deletions in the T-cell receptor delta locus of mouse thymocytes.

Molecular and Cellular Biology ◽

10.1128/mcb.14.7.4455 ◽

1994 ◽

Vol 14 (7) ◽

pp. 4455-4464 ◽

Cited By ~ 9

Author(s):

S M Fish ◽

M J Bosma

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Dna Sequences ◽

Cell Receptor ◽

Antigen Receptors ◽

Wild Type ◽

Variable Regions ◽

Age Dependent ◽

A Site ◽

Signal Coding

Separate genetic elements (V, D, and J) encode the variable regions of lymphocyte antigen receptors. During early lymphocyte differentiation, these elements rearrange to form contiguous coding segments (VJ and VDJ) for a diverse array of variable regions. Rearrangement is mediated by a recombinase that recognizes short DNA sequences (signals) flanking V, D, and J elements. Signals flank both the 5' and 3' sides of each D element, thereby allowing assembly of a functional VDJ gene. However, in rearrangements involving the D delta 2 and J delta 1 elements of the mouse T-cell receptor delta (TCR delta) locus, we unexpectedly found that the D delta 2 element and a portion of its 5' signal are often deleted. Approximately 50% of recovered D delta 2 to J delta 1 rearrangements from thymocytes of adult wild-type mice showed such deletions. An additional 20% of the rearrangements contained standard D delta 2-J delta 1 coding junctions but showed some loss of nucleotides from the 5' D delta 2 signal. This loss was clearly associated with another event involving a site-specific cleavage at the 5' signal/coding border of D delta 2 and rejoining of the modified signal and coding ends. The abnormal loss of D delta 2 and a portion of the 5' D delta 2 signal was infrequently observed in D delta 2-to-J delta 1 rearrangements recovered from neonatal mice. The possible basis and significance of this age-dependent phenomenon are discussed.

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