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By MedGenome Scientific Affairs
The field of immune repertoire profiling has witnessed remarkable advancements in recent years, revolutionizing our understanding of the immune system and its role in various diseases. One of the key techniques to understand this complex mechanism is TCR sequencing. TCR, or T-cell receptor, plays a crucial role in the adaptive immune response by recognizing and binding to specific antigens. By sequencing the TCR repertoire, researchers can gain valuable insights into the diversity and specificity of T-cell populations, leading to the development of novel treatment modalities in infectious diseases, autoimmunity, and immuno-oncology.
TCR sequencing has proven to be a powerful tool for understanding:
- Host-pathogen interactions and designing effective therapeutic strategies: Through analysis and identification of specific T-cell clones associated with protective immune responses, novel vaccines or immunotherapies that target these specific T-cell clones can be developed.
- TCR sequencing provides crucial insights into the underlying mechanisms of Auto-immune disease development: By comparing the TCR repertoires of patients with autoimmune disorders to those of healthy individuals, researchers have been able to identify aberrant T-cell populations that are associated with autoimmune pathology. This knowledge aids in the development of targeted therapies that restore immune balance and alleviate autoimmune symptoms.
- TCR sequencing holds immense promise for personalized cancer treatment: By profiling the TCR repertoire of tumor-infiltrating lymphocytes (TILs), researchers can identify T-cell clones that are specifically targeting tumor antigens. This information can then be used to engineer T-cell-based immunotherapies, such as chimeric antigen receptor (CAR) T-cell therapy, that specifically target and eliminate cancer cells. TCR sequencing also allows for the monitoring of treatment response and the identification of potential immune escape mechanisms employed by tumors.
RNA based TCR repertoire profiling
While traditional TCR sequencing methods rely on genomic DNA, recent advancements in RNA-based sequencing techniques have expanded the scope of immune repertoire profiling. RNA-based TCR repertoire profiling offers several advantages over DNA-based methods, providing deeper insights into T-cell dynamics and functionality.
By profiling the TCR repertoire at the RNA level, researchers can capture the transcriptomic landscape of T-cells, allowing for the identification of actively expressed T-cell clones. This enables a more accurate representation of the T-cell diversity and functionality within a given sample. Moreover, RNA-based TCR sequencing facilitates the characterization of antigen-specific T-cells, enabling researchers to map the immune response to specific antigens with greater precision.
RNA-based TCR repertoire profiling also allows for the detection of alternative splicing events within the TCR transcripts. Alternative splicing can result in the generation of T-cell receptor isoforms with distinct antigen-binding properties. By capturing these isoforms, researchers can gain a deeper understanding of T-cell receptor diversity and its implications for immune recognition and response.
BCR repertoire profiling in diagnostic biomarker discovery and disease diagnosis
B-cells, through their B-cell receptors (BCRs), play a crucial role in humoral immunity by recognizing and binding to antigens. Profiling the BCR repertoire offers valuable insights into B-cell differentiation, BCR somatic hypermutation, class switching, and antigen specificity.
One of the key applications of BCR repertoire profiling is in diagnostic biomarker discovery. By analyzing the BCR repertoires of patients with certain diseases, researchers can identify disease-specific B-cell clones or antibody sequences. These disease-associated BCR sequences can then be utilized as diagnostic biomarkers, aiding in the early detection and monitoring of diseases. Furthermore, BCR repertoire profiling can also provide insights into disease progression and treatment response, enabling personalized medicine approaches.
BCR repertoire profiling provides a better picture for disease diagnosis. By comparing the BCR repertoires of healthy individuals to those of patients, researchers can identify disease-specific B-cell clones or antibody sequences. This information can aid in the early diagnosis and classification of diseases, facilitating timely interventions and improving patient outcomes.
Deeper insights into B-cell differentiation, BCR somatic hypermutation, class switching, and antigen specificity
Beyond its applications in biomarker discovery and disease diagnosis, BCR repertoire profiling provides deeper insights into various aspects of B-cell biology. By analyzing the BCR repertoires of different B-cell subsets, researchers can unravel the intricate processes of B-cell differentiation. This knowledge not only enhances our understanding of normal immune development but also sheds light on the dysregulation of B-cell differentiation in diseases such as leukemia and lymphoma.
BCR repertoire profiling is also instrumental in studying BCR somatic hypermutation and class switching. Somatic hypermutation is a key mechanism through which B-cells generate high-affinity antibodies, while class switching allows to produce different antibody isotypes with distinct effector functions. By analyzing the BCR repertoires of B-cell subsets at different stages of somatic hypermutation and class switching, researchers can decipher the underlying molecular mechanisms and regulatory networks governing these processes.
Furthermore, BCR repertoire profiling enables the characterization of antigen-specific B-cell populations. By identifying B-cell clones that are enriched for specific antigen-binding sequences, researchers can gain insights into the antigen-specific immune response. This information can be valuable for vaccine development, as it helps in the identification of immunogenic epitopes and the assessment of vaccine efficacy.
In conclusion, immune repertoire profiling, particularly through TCR and BCR sequencing, has revolutionized our understanding of the immune system and its role in various diseases. From infectious diseases to autoimmunity and immuno-oncology, TCR sequencing has paved the way for novel treatment modalities. RNA-based TCR repertoire profiling offers deeper insights into T-cell dynamics and functionality. On the other hand, BCR repertoire profiling provides valuable information about B-cell differentiation, somatic hypermutation, class switching, and antigen specificity. By harnessing the power of immune repertoire profiling, we are unlocking new frontiers in diagnostics, therapeutics, and our overall understanding of the immune system.
At MedGenome, we provide TCR and BCR repertoire profiling using bulk input (from cells, RNA) using the SMARTerTCR/BCR Profiling Kit (Takara Bio USA Inc) the Chromium Immune Profiling solutions (10X Genomics). We have expertise in processing a variety of sample types at high-throughput mode.
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- Complete workflow — Sample extraction, library prep, sequencing and for seamless data analysis and visualization
- Increased accuracy — unique molecular identifier (UMI)-based PCR error correction
- Detection of TCR clonotypes — Detection of novel clonotypes, and sensitive identification of full-length V(D)J and gives high resolution TCR-α and TCR-β pairing information.
- Detection of all BCR isotypes — sequence all heavy and light chains seamlessly with pooled primers
References
Shugay M. et al. Towards error-free profiling of immune repertoires. Nat. Methods 11, 653–655 (2014).
Yaari, G. and Kleinstein, S.H. Practical guidelines for B-cell receptor repertoire sequencing analysis. Genome Med. 7:121 (2015).
Georgiou, G., Ippolito, G., Beausang, J. et al. The promise and challenge of high-throughput sequencing of the antibody repertoire. Nat Biotechnol 32, 158–168 (2014).
Six, A., et al. (2013) The past, present, and future of immune repertoire biology–the rise of next-generation repertoire analysis. Front. Immunol. 4(413):1–16.
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