Identification of Human T Cell Receptor γδ-recognized Epitopes/Proteins via CDR3δ Peptide-based Immunobiochemical Strategy

Chen, Hui; He, Xiancong; Wang, Zhun; Wu, Di; Zhang, Huiyuan; Xu, Chunping; He, Haixiang; Cui, Lianxian; Ba, Denian; Wang, He

doi:10.1074/jbc.m708067200

Cited by 59 publications

(64 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With this novel strategy, we have successfully identified seven tumor-related epitopes, two hepatitis B virus (HBV) infection-related antigenic epitopes, and two self proteins including heat shock protein (HSP) 60 and human mutS homolog 2 (hMSH2) that are recognized by human ␥␦ TCR (7). These results further support that the primary sequence of CDR3␦ in ␥␦ TCR determine the specificity of antigen binding.…”

supporting

confidence: 59%

“…Nine peptides as putative epitopes were identified, among which seven were tumor-related and two were HBV infection-related. BLAST searches revealed that most matched proteins were conserved proteins of prokaryotes (7). Results suggest the possibility that ␥␦ TCR recognizes some conserved molecules.…”

Section: Discussionmentioning

confidence: 88%

“…We mutated V, N-D-N, or J fragments of a V␦2 TCR CDR3 sequence (OT3) in peptide and engineered ␥␦ TCR. We found that the conserved flanking regions of CDR3␦ play a critical role in antigenic binding to OEC cells/tissues or hMSH2 protein, a new ligand for ␥␦TCR we found recently (7). Furthermore, we have identified the cysteine residue in V fragment and the leucine residue in J fragment as critical residues in the binding activity of ␥␦ TCR.…”

mentioning

confidence: 77%

See 2 more Smart Citations

Antigen Specificity of γδ T Cells Depends Primarily on the Flanking Sequences of CDR3δ

Xi¹,

Guo²,

Chen

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The structural basis that determines the specificity of ␥␦ T cell receptor (TCR) recognition remains undefined. Our previous data show that the complementary determining region of human TCR␦ (CDR3␦) is critical to ligand binding. Here we used linear and configurational approaches to examine the roles of V, N-D-N, or J regions in CDR3␦-mediated antigen recognition. Surprisingly, we found that the binding activities of CDR3␦ from different ␥␦ TCRs to their target tissues and ligands depend on the conserved flanking sequences (V and J) but not as much on the D region of CDR3␦ fragment. We further defined the key residues in the V and J regions of CDR3␦ fragments, including the cysteine residue in the V fragment and the leucine residue in the J fragment that determine their ligand binding specificity. Our results demonstrate that TCR␦ primarily uses conserved flanking regions to bind ligands. This finding may provide an explanation for the limited number of ␥␦ TCR ligands that have as yet been identified.Extensive studies suggest that ␥␦ T cells play important roles in host defense against microbial infections, monitoring of tumorigenesis, immunoregulation, and development of autoimmunity (1-3). However, little is known about the structural basis of antigenic recognition by ␥␦ T cell receptor (TCR) 3 because of the limited identified specific ligands for ␥␦ TCR and the lack of structural information revealing how ␥␦ TCR might interact with such ligands.The crystallographic structure of a murine ␥␦ TCR in complex with major histocompatibility complex class (MHC) Ib T22 (4, 5) showed that the CDR loops f ␥␦ TCR, predominantly germline-encoded residues of the complementary determining region of human TCR␦ (CDR3␦), are in direct contact with T22, suggesting that the primary sequence of CDR3 in ␥␦ TCR, especially CDR3␦, serves as a key determinant for the specificity of antigen recognition. Our recent finding that CDR3␦ peptide mimics human ␥␦ TCR binding to tumor cells and tissues is consistent with the role of CDR3␦ in ␥␦ TCR recognition (6).Based on this finding, we used synthesized CDR3␦ peptide as a probe to screen putative protein ligands in tumor protein extracts by affinity chromatography analysis. With this novel strategy, we have successfully identified seven tumor-related epitopes, two hepatitis B virus (HBV) infection-related antigenic epitopes, and two self proteins including heat shock protein (HSP) 60 and human mutS homolog 2 (hMSH2) that are recognized by human ␥␦ TCR (7). These results further support that the primary sequence of CDR3␦ in ␥␦ TCR determine the specificity of antigen binding.CDR3␦ is composed of fragments derived from V, N-D-N, and J gene segments. The flanking sequences composed of V and J fragment is conserved while N-D-N region is diverse. The diversity of ␥␦ TCRs is supposedly higher than that of TCR␣␤ due to the link of D gene fragment and the insertion of nucleotide acids (8). However, the number of identified antigenic ligands recognized by ␥␦ TCR remains very limited. It has been d...

show abstract

supporting

confidence: 59%

Section: Discussionmentioning

confidence: 88%

mentioning

confidence: 77%

See 1 more Smart Citation

Antigen Specificity of γδ T Cells Depends Primarily on the Flanking Sequences of CDR3δ

Xi¹,

Guo²,

Chen

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…76 More recently, another putative Vc9Vd2 T cell tumor antigen, human MutS homologue 2 (MSH2), was identified using a peptide-based affinity screening system for the CDR3 of the d chain. 77 MSH2 is normally located in the nucleus where it functions as a DNA mismatch repair gene, but it is often mutated in a number of different types of epithelial cancers and can be ectopically expressed. 78 Transformation of normal human B cells by Epstein-Barr virus or subjecting renal carcinoma cell lines to oxidative stress also led to an increased surface expression of MSH2 and rendered them susceptible to Vc9Vd2 T cell-mediated cell lysis that could be blocked by the use of anti-MSH2 antibodies or downregulation of its gene expression.…”

Section: Vd2 T Cells: To Know the Complex Burden Of Being Humanmentioning

confidence: 99%

Defining the nature of human γδ T cells: a biographical sketch of the highly empathetic

2012

View full text Add to dashboard Cite

The elusive task of defining the character of cd T cells has been an evolving process for immunologists since stumbling upon their existence during the molecular characterization of the a and b T cell receptor genes of their better understood brethren. Defying the categorical rules used to distinctly characterize lymphocytes as either innate or adaptive in nature, cd T cells inhabit a hybrid world of their own. At opposing ends of the simplified spectrum of modes of antigen recognition used by lymphocytes, natural killer and ab T cells are particularly well equipped to respond to the 'missing self' and the 'dangerous non-self', respectively. However, between these two reductive extremes, we are chronically faced with the challenge of making peace with the 'safe non-self' and dealing with the inevitable 'distressed self', and it is within this more complex realm cd T cells excel thanks to their highly empathetic nature. This review gives an overview of the latest insights revealing the unfolding story of human cd T cells, providing a biographical sketch of these unique lymphocytes in an attempt to capture the essence of their fundamental nature and events that influence their life trajectory. What hangs in their balance is their nuanced ability to differentiate the friends from the foe and the pathological from the benign to help us adapt swiftly and efficiently to life's many stresses. Keywords: antigen recognition; danger-associated molecular patterns; gamma delta T cell subsets; immune homeostasis; innate immunity; stress response PROLOGUE Since the time of their accidental discovery during the molecular characterization of the a and b T cell receptor (TCR) genes belonging to their relatively unambiguous brethren, 1 'enigmatic' is still one of the most commonly used adjectives to describe cd T cells. This can be attributed to the fact that these unconventional lymphocytes have thus far been remarkably successful in thwarting most attempts at definition. Much of what we know about T cell biology and function comes from what has been discerned through studies done on ab T cells; however, one thing that is certain is that the majority of the rules governing the lives of ab T cells are surprisingly irrelevant when it comes to understanding the elusive character of cd T cells. Table 1 highlights some of the major differences between these two T cell subsets in humans.Recognition by cd T cells is not typically in the context of classical major histocompatibility complex (MHC) class I or II molecules and neither is antigen processing required. 2,3 These observations are corroborated by the fact that the majority of cd T cells are CD8 and CD4 negative. Crystal structure analysis of the cd TCR determined that the length and conformation of cd TCR resemble immunoglobulins (Ig) more than the ab TCR, which was taken to suggest that antigen recognition by cd T cells may be more similar to the binding of antibody to antigen rather than the MHC/peptide complex recognized by ab T cells. 4 The very basis of foreign antigen rec...

show abstract

“…We found that synthesized OEC-derived CDR3d-peptides could mimic the TCRcd and bind specifically to tumor cell lines and tissues 25 A CDR3d-grafted antibody also showed specific binding to several tumor cells 19 We used synthesized OEC-derived CDR3d peptides as probes to search for putative TCR-binding epitopes by screening a 12-mer random peptide phage-displayed library and identified several putative TCR ligands within tumor cell extracts by affinity chromatography and liquid chromatography/electrospray ionization tandem mass spectrometry analysis. Nine peptides and two proteins that bound the cdTCR been identified including human mutS homolog 2, a novel cdTCR ligand 15,31 Another research group also found that the CDR3d region was responsible for the T22 protein binding specificity of the Vd2 T cells clone G8 32 We had previously constructed full-length PBMCs-derived c9 and d2 chains using overlapping PCR. The fulllength c9 and d2 chains with tumor antigen-specific CDR3 regions were stably transfected into J.RT3-T3.5 by electroporation and expressed functional c9d2TCR 26 In this study, we induced c9 and d2 (OT3) expression in PBLs using a retroviral vector and generated efficient tumor reactive T cells.…”

Section: Discussionmentioning

confidence: 99%

CDR3δ -grafted γ9δ2T cells mediate effective antitumor reactivity

Zhao

Cui

et al. 2011

Cell Mol Immunol

Self Cite

View full text Add to dashboard Cite

Adoptive cell-transfer therapy (ACT) has been reported to suppress growing tumors and to overcome tumor escape in animal models. As a candidate ACT effector, c9d2T cells can be activated and expanded in vitro and in vivo and display strong antitumor activity against colorectal, lung, prostate, ovarian and renal cell carcinomas. However, it is difficult to obtain a large enough number of cdT cells to meet the need for immunotherapy that can overcome the cancer patients' immune suppressive tumor microenvironment. In previous studies, our lab confirmed that c9d2T cells recognized tumor cells via the CDR3d region of the cd-T-cell receptor (TCR). We constructed full-length human peripheral blood mononuclear cell (PBMC)-derived c9 and d2 chains in which the CDR3 region was replaced by an ovarian epithelial carcinoma (OEC)-derived CDR3. We transferred the CDR3d-grafted c9d2TCR into peripheral blood lymphocytes (PBLs) to develop genetically modified c9d2T cells. In vitro studies have shown that these CDR3d-grafted c9d2T cells can produce cytokines after stimulation with tumor cell extracts and exhibit cytotoxicity towards tumor cells, including human OEC and cervical adenocarcinoma. CDR3d-grafted c9d2T cells adoptively transferred into nude mice bearing a human OEC cell line demonstrated significant antitumor effects. These results indicate that CDR3d-grafted c9d2T cells might be candidates for clinical tumor immunotherapy.

show abstract

Identification of Human T Cell Receptor γδ-recognized Epitopes/Proteins via CDR3δ Peptide-based Immunobiochemical Strategy

Cited by 59 publications

References 34 publications

Antigen Specificity of γδ T Cells Depends Primarily on the Flanking Sequences of CDR3δ

Antigen Specificity of γδ T Cells Depends Primarily on the Flanking Sequences of CDR3δ

Defining the nature of human γδ T cells: a biographical sketch of the highly empathetic

CDR3δ -grafted γ9δ2T cells mediate effective antitumor reactivity

Contact Info

Product

Resources

About