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TCR/BCR Repertoire Analysis

Diversity analysis of immune cells involved in specific antigen recognition will be performed at the gene level. High quality repertoire analysis is being implemented by optimized methods such as unbiased amplification, next generation sequencing, and dedicated bioinformatics.

Neoepitope Analysis

By conducting RNA seq and Exose seq together, specific gene mutations which are specific to individual cancers can be identified. In addition, by taking into account intermolecular coupling, variant peptides (neoepitope) capable of presenting an antigen will be identified. This identification can also be used for indel, long indel, and splicing variants using sequence data.

16S rRNA Bacterial Flora Analysis

Distribution of bacterial species (bacterial flora) in samples can be analyzed by examining the V1V2 or V3V4 region of the 16S rRNA gene preserved in each bacterial species using next generation sequencing. This analysis can be applied not only to biologically derived samples (feces, saliva, etc.), but also to environmentally derived samples (soil, food, etc.).

Publications

2024/07/22
Paper

The Journal of Dermatology

Involvement of the genus Corynebacterium in the pathogenesis of pigmented intratarsal keratinous cyst

A paper utilizing our 16S rRNA Bacterial Flora Analysis was published by Dr. Miyuki Yoshikawa, Dr. Daiki Rokunohe et al., Hirosaki University Graduate School of Medicine, Department of Dermatology, has published a paper on inflammation and bacterial flora in intratarsal keratinous cysts (IKC), which are pigmented intraocular keratinocysts.
IKC, a benign cystic lesion of the eyelid, usually appears as a yellow to white cyst, but rarely as a brown or gray-blue cyst, affecting the clinical diagnosis. The cause of this phenomenon is not known. The author's research group has observed a localized lymphocytic infiltrate in the melanocyte-rich, melanin-positive area below the cyst wall in pigmented IKC, and has analyzed the bacterial colonies within the cyst for the presence of Corynebacterium species. The etiology of pigmented IKC related to inflammation and bacterial flora was discussed in the paper.
We have performed 16S rRNA Bacterial Flora Analysis using the bacterial colonies in the cysts described in the paper.

2024/07/22
Paper

Pathology International

Solid tumor
Regulatory T-cells activated in metastatic draining lymph nodes possibly suppress cancer immunity in cancer tissues of head and neck squamous cell cancer

A paper utilizing our TCR Repertoire Analysis was published by Dr. Susumu Suzuki et al., Aichi Medical University Research Creation Support Center that suggests regulatory T cells activated in metastatic lymph nodes suppress cancer immunity in head and neck squamous cell carcinoma (HNSCC) cancer tissue.

Since the mechanism by which activated regulatory T cells (Tregs) suppress cancer immunity is unknown, to elucidate this mechanism, the research group performed T cell receptor (TCR) repertoire analysis was performed. We found that the TCR repertoires were biased in cancer tissue and metastatic DLNs (M-DLNs) compared to non-metastatic DLNs, and that the TCR repertoires of Tregs and CD8+ T cells between M-DLNs and cancer tissue were more similar than in other sites. These results suggest that Treg and CD8+ T cells are activated by cancer antigens such as neoantigens and shared antigens in M-DLNs and cancer tissues, and that Tregs suppress CD8+ T cell function in a cancer antigen-specific manner in M-DLNs and cancer tissues. Furthermore, M-DLN may be a source of Tregs and CD8+ T cells that are recruited to cancer tissues. These findings suggest that antigen-specific targeting of Tregs with M-DLN may represent a novel immunotherapeutic strategy for squamous cell carcinoma of the head and neck.

We have performed a TCR Repertoire Analysis using Tregs and conventional T cells in peripheral blood, influx regional lymph nodes (DLNs), and cancer tissues of patients with squamous cell carcinoma of the head and neck (HNSCC) as shown in the article.

2024/07/01
Paper

Cell Stem Cell

Basal immunity
Human iPSC-derived CD4+ Treg-like cells engineered with chimeric antigen receptors control GvHD in a xenograft model

A paper utilizing our TCR Repertoire Analysis was published by Dr. Hisashi Yano, Shin Kaneko Laboratory, CiRA, Kyoto University.
In this paper, they report a success in producing CD4+ Treg-like cells derived from human iPS cells by inducing FOXP3 expression.
Using our TCR Repertoire Analysis, it was confirmed that cells cultured from non-T cell-derived iPS cells reconstitute various TCRα and β genes during their differentiation, constituting a polyclonal TCR repertoire.

TCR/BCR Repertoire analysis Guidebook

MUST READ! For users considering Repertoire analysis

An easy-to-understand repertoire analysis guidebook has been prepared which explains in plain language for the questions such as what is immune repertoire analysis and what kind of preparation is necessary.
Please request the guidebook from below if desired.

TCR/TCR/BCRレパトア解析ガイドブック

Number of pages:71, File size: 9.5 MB, File format: PDF

About the structure of this guidebook

Chapter 1: TCR/BCR Repertoire

  • 1)Overview
  • 2)Somatic Recombination
  • 3)Somatic Hypermutation
  • 4)Scale of Diversity
  • 5)Differences in gDNA and mRNA after Somatic Recombination

Chapter 2:
Analysis of Repertoire

  • 1) Overview
  • 2) History 1 (Until the 20th Century)
  • 3) History 2 (Appearance of NGS)
  • 4) History 3 (Appearance of Long Read Sequencer)
  • 5) History 4 (Reliability Assurance and Automation)《New!》
  • 6) Gene Amplification Method
  • 7) Advantages and Disadvantages
  • 8) Risks of PCR Bias
  • 9) Risk of Primer Mismatch in Multiplex PCR is Impossible
  • 10) Determination of the Isotype with Multiplex PCR
  • 11) Versatility of Repertoire Analysis and its Application to Various Animal Species《New!》
  • 12) "Pair Identification" of Repertoire Using Single-Cell Analysis《New!》
  • 13) Full-Length Prediction Analysis《New!》
  • 14) Possibility of Antigen Predictive Analysis《New!》

Chapter 3:
Sample Handling

  • 1) Overview of Analyzable Samples and Handling
  • 2) Handling of Whole Blood Samples
  • 3) Separation of PBMC from Blood (Whole Blood) Samples
  • 4) Handling of Isolated Cell Samples
  • 5) Handling of Tissue Samples
  • 6) Handling of Total RNA Samples

Chapter 4:
Interpretation of Repertoire Analysis Data

  • 1)Overview
  • 2)Interpretation of Repertoire Analysis Results
  • 3)About Supplementary Material
  • 4)Frequently Asked Questions and Answers

Chapter 5:
Publications Using Repertoire Analysis《New!》

  • 1)List of Publications (62 reports)
  • 2)Publication Summary

Chapter 6:
Analysis Service Lineup (Digest Version)《New!》

  • - Overview and Basic Request Procedures
  • - TCR/BCR Repertoire Analysis
  • - Neoepitope Analysis
  • - 16S rRNA Bacterial Flora Analysis

Customer Feedback

University professor A

The realization of personalized medicine requires the identification of tumor antigens and cytotoxic T cell therapy to target tumor antigens. Neoepitope search・identification software and TCR repertoire analysis provided by Repertoire Genesis Inc. made it possible to identify targets and corresponding TCR for subsequent application to gene therapy, enabling dendritic cell vaccination therapy. One can truly feel that this marks the beginning of cutting-edge medicine.

University professor B
Field:Hematological oncology

By TCR/BCR repertoire analysis, various clinical issues can be solved. TCR/BCR repertoire analysis is considered necessary for evaluating and predicting the success or failure of bone marrow transplantation in our lab; thus, we request repertoire analysis frequently. Moreover, this analysis is highly useful for clinical diagnosis and determination of treatment efficacy. Neoepitope analysis recently developed by Repertoire Genesis Inc. can be applied widely in the hematology field and it is believed that the analysis can be quickly used for predicting therapy effect for CML. It is a technology progressively increasing clinical knowledge with extremely broad scope of application.

University professor C
Field:Hematological oncology

Our lab is carrying out studies of infectious disease, allergy, and autoimmune disease based on hematology and immunology, pursuing diagnostic methods which can be applied across organs. Within our research, determining whether the immune system is specifically involved in disease using TCR/BCR repertoire analysis developed by Repertoire Genesis, Inc. plays an extremely important role. Moreover, we think that the cause of idiopathic purpura will be elucidated by TCR/BCR repertoire analysis.

Director of a research institution D
Field:Virology

By TCR/BCR repertoire analysis, the research field of infection immunity is thought to progress dramatically. Until recently, the importance of T cell involvement has been elucidated using a virus infected animal model. In the future, we expect to apply BCR analysis for developing antibody therapy drugs. Thus, continuously analyses are requested to Repertoire Genesis, Inc.

University professor E
Field:Immunology

To study the relationship between lifestyle diseases (obesity) and the immune system, we are using an arteriosclerosis animal model for analysis. We consider TCR/BCR repertoire analysis to be the most effective tool for conducting precise characterization analysis of cells infiltrating the arteriosclerotic region.
Analysis results by Repertoire Genesis Inc. showed that involvement of NKT cells in lifestyle diseases is strongly indicated. Research from a perspective of relationship between lifestyle diseases (obesity) and the immune system may seem weak at first. However, the relationship between many diseases and the immune system will be elucidated by technologies provided by Repertoire Genesis Inc. Thus, progress in future research is anticipated.

University professor F
Field:Hematological oncology

The fatality rate of ATL found in Kyushu area is extremely high after onset. In recent years, success in treating ATL by opdivo has been confirmed. However, the mode of action for drug efficacy is not clear.
TCR repertoire analysis by Repertoire Genesis Inc. revealed the involvement between the hemogram pattern in peripheral blood and Treg after administration of opdivo. Therefore, the analysis results may bring some good news in the future. Moreover, it appears that a final confirmation of ATL metastasis to the skin will be performed by TCR repertoire. It is also thought that many cases of leukemia and lymphoma will require TCR/BCR repertoire analysis. We intend to continue using the service provided by Repertoire Genesis Inc.

University professor G (overseas)
Field:Immunology

Now we would have extreme use of Repertoire Genesis's sequencing technology because sequencing T cells on a massive scale would provide more useful representation of the TCR repertoire and provide a fundamental basis for progressing to more detailed cell culture/in vivo experimentation.

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