Identification of Tumoricidal TCRs from Tumor-Infiltrating Lymphocytes by Single-Cell Analysis

Shitaoka, Kiyomi; Hamana, Hiroshi; Kishi, Hiroyuki; Hayakawa, Yoshihiro; Kobayashi, Eiji; Sukegawa, Kenta; Piao, Xiuhong; Lyu, Fulian; Nagata, Takuya; Sugiyama, Daisuke; Nishikawa, Hiroyoshi; Tanemura, Atsushi; Katayama, Ichiro; Murahashi, Mutsunori; Takamatsu, Yasushi; Tani, Kenzaburo; Ozawa, Tatsuhiko; Muraguchi, Atsushi

doi:10.1158/2326-6066.cir-17-0489

Cited by 30 publications

(45 citation statements)

References 39 publications

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“…However, a direct functional assay of TCRs obtained from Treg cells is still not available, reflecting the limited knowledge about epitopes from physiological targets of Treg cells (27, 36). TCRs from tumor specific conventional CD4 + T cells and CD8 + T cells were identified using single-cell RT-PCR methods (23, 24, 37, 38). We also demonstrated a rapid and efficient functional assay for TCRs directly obtained from CD8 + T cells using TCR-transduced lymphocytes in the absence of information on antigen or MHC haplotype (38).…”

Section: Discussionmentioning

confidence: 99%

“…TCRs from tumor specific conventional CD4 + T cells and CD8 + T cells were identified using single-cell RT-PCR methods (23, 24, 37, 38). We also demonstrated a rapid and efficient functional assay for TCRs directly obtained from CD8 + T cells using TCR-transduced lymphocytes in the absence of information on antigen or MHC haplotype (38). We tried to apply this technique to mixed lymphocyte reaction of paired αβ TCR-transduced maternal T cells or TCR-negative T cell lines against self-fetal umbilical cord blood but were unsuccessful.…”

Section: Discussionmentioning

confidence: 99%

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Clonally Expanded Decidual Effector Regulatory T Cells Increase in Late Gestation of Normal Pregnancy, but Not in Preeclampsia, in Humans

et al. 2018

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Background: Regulatory T (Treg) cells are necessary for the maintenance of allogenic pregnancy. However, the repertoire of effector Treg cells at the feto-maternal interface in human pregnancy remains unknown. Our objective was to study T cell receptor (TCR) repertoires of Treg cells during pregnancy compared to normal and complicated pregnancies.Methods:Paired samples of peripheral blood and decidua in induced abortion and miscarriage cases were obtained from consenting patients. CD4+CD25+CD127low/−CD45RA− effector Treg cells were single-cell sorted from mononuclear cells. cDNAs of complementarity determining region 3 (CDR3) in TCRβ were amplified from the single cells by RT-PCR and the sequences were analyzed. The TCRβ repertoires were determined by amino acid and nucleotide sequences. Treg cells were classified into clonally expanded and non-expanded populations by CDR3 sequences.Results: We enrolled nine induced abortion cases in the 1st trimester, 12 cases delivered without complications in the 3rd trimester, 11 miscarriages with abnormal chromosomal karyotyped embryo, seven miscarriages with normal chromosomal karyotyped embryo, and seven cases of preeclampsia [median gestational week (interquartile range): 7 (7–9), 39 (38–40), 9 (8–10), 8 (8–10), and 34 (32–37), respectively]. The frequency of clonally expanded populations of effector Treg cells increased in decidua of 3rd trimester cases compared to 1st trimester cases [4.5% (1.4–10.8%) vs. 20.9% (15.4–28.1%), p < 0.001]. Clonally expanded Treg cells were rarely seen in peripheral blood. The ratio of clonally expanded populations of decidual effector Treg cells in miscarriages with abnormal and normal embryos was not significantly different compared with that in 1st trimester normal pregnancy. Interestingly, clonally expanded populations of effector Treg cells decreased in preeclampsia compared with that in 3rd trimester normal pregnancy [9.3% (4.4–14.5%) vs. 20.9% (15.4–28.1%), p = 0.003]. When repertoires in previous pregnancy and subsequent pregnancy were compared, some portions of the repertoire were shared.Conclusion: TCR repertoires of decidual effector Treg cells are skewed in the 3rd trimester of normal pregnancy. Failure of clonal expansion of populations of decidual effector Treg cells might be related to the development of preeclampsia.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Clonally Expanded Decidual Effector Regulatory T Cells Increase in Late Gestation of Normal Pregnancy, but Not in Preeclampsia, in Humans

et al. 2018

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“…Importantly, the transduced T cells expressing TCRs recognized the neoantigen presented by autologous antigen-presenting cells [124] . Another study using single-cell TCR repertoire analysis revealed that clonally expanded CD8 T cells were antigen-specific and showed cytotoxic activity against tumors in mouse models [126] . Intriguingly, the combination of 10x Genomics' single cell TCR sequencing platform coupled to gene expression holds enormous potential for assessing and monitoring patient response to cancer vaccines and immunotherapy drugs.…”

Section: Tcr Repertoire Analysismentioning

confidence: 99%

Unmasking tumor heterogeneity and clonal evolution by single-cell analysis

Shi¹,

Chakraborty²,

Chaudhuri³

2018

JCMT

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The intratumoral heterogeneity orchestrated by the tumor intrinsic and extrinsic mechanisms enable cancers to persist and spread notwithstanding the use of aggressive interventional therapies. The heterogeneity is revealed at multiple levels-at the level of individual tumor cells, in the cellular composition of tumor infiltrates and in the chemical microenvironment in which the cells reside. Deconvoluting the complex nature of the cell types present in the tumor, along with the homo and heterotypic interactions between different cell types can produce novel insights of biological and clinical relevance. However, most techniques analyze tumors at a gross level missing key inter-cell-type genotypic and phenotypic differences. The advent of single-cell sequencing has given an unprecedented opportunity to analyze the tumor at a resolution that not only captures the diversity of the cellular composition of a tumor but also provides information on the genetic, epigenetic and functional states of different cell types. In this review, we summarize the genesis of tumor heterogeneity, its impact on tumor growth and progression and their clinical consequences. We present an overview of the currently available platforms for isolation and sequencing of single tumor cells and provide evidence of its utility in precision medicine and personalized therapy.

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“…Clonally expanded TCRαβ+ T cells were identified in patients with hematologic malignancies and cancer, and the identified tumor antigen‐specific TCRαβ gene could be used to construct TCR‐modified T cells (TCR‐T) for cancer immunotherapy . Molecular analysis, particularly using next‐generation sequencing and single‐cell sequencing, could identify clonal TCR rearrangements, confirming specifically expanded T cell clones and monitoring antigen‐specific T cell responses, which may help to design effective TCR‐based immunotherapies . However, these analyses are incapable of evaluating the functional status of clonally expanded T cells.…”

Section: Introductionmentioning

confidence: 99%

“…2,[25][26][27] Molecular analysis, particularly using next-generation sequencing and single-cell sequencing, could identify clonal TCR rearrangements, confirming specifically expanded T cell clones and monitoring antigen-specific T cell responses, which may help to design effective TCR-based immunotherapies. [28][29][30] However, these analyses are incapable of evaluating the functional status of clonally expanded T cells. Knowledge of the distribution of the TCR + subfamily in T cell subsets in patients with leukemia is lacking.…”

Section: Introductionmentioning

confidence: 99%

A skewed distribution and increased PD-1+Vβ+CD4+/CD8+ T cells in patients with acute myeloid leukemia

Huang

Tan

Chen

et al. 2019

Journal of Leukocyte Biology

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The limited application of immunotherapy in acute myeloid leukemia (AML) may be due to poor understanding of the global T cell immune dysfunction in AML. In this study, we analyzed the distribution characteristics of 24 TCR Vβ subfamilies in CD3+, CD4+, and CD8+ T cells in AML patients and healthy controls. The percentage of TCR Vβ subfamily T cells was predominately lower in most AML cases, while it was increased in some cases. TCR Vβ2+T cells were increased in AML, particularly TCR Vβ2+CD4+T cells, which were significantly higher. To further address the immunosuppression in different Vβ subfamilies, we characterized the distribution of program death‐1 (PD‐1)+T cells in TCR Vβ subfamilies of CD4+ and CD8+T cells. Significantly higher levels of PD‐1+Vβ+T cells were found for most Vβ subfamilies in most AML cases. A higher percentage of PD‐1+Vβ2+T cells with a high number of Vβ2+T cells was found in all of the CD3+, CD4+, and CD8+ T cell subsets. Moreover, increasing PD‐1+Vβ7.2, Vβ8+, Vβ14+, Vβ16+, and Vβ22+CD8+T cells were distributed in the AML‐M5 subtype group compared with the AML‐M3 group. In addition, higher PD‐1+ Vβ5.2+ and PD‐1+ Vβ12+CD8+T cells were associated with AML patients who had a poor response to chemotherapy. In conclusion, increased PD‐1+Vβ+T cells is a common characteristic of AML, higher PD‐1+Vβ2+T cells may be associated with a low antileukemia effect, and higher PD‐1+Vβ5.2+ and PD‐1+Vβ12+CD8+T cells may be related to poor prognosis in AML. These characteristics may be worth considering as immune biomarkers for clinical outcome in AML.

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Identification of Tumoricidal TCRs from Tumor-Infiltrating Lymphocytes by Single-Cell Analysis

Cited by 30 publications

References 39 publications

Clonally Expanded Decidual Effector Regulatory T Cells Increase in Late Gestation of Normal Pregnancy, but Not in Preeclampsia, in Humans

Clonally Expanded Decidual Effector Regulatory T Cells Increase in Late Gestation of Normal Pregnancy, but Not in Preeclampsia, in Humans

Unmasking tumor heterogeneity and clonal evolution by single-cell analysis

A skewed distribution and increased PD-1+Vβ+CD4+/CD8+ T cells in patients with acute myeloid leukemia

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