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Single Cell Immune Profiling

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Single Cell Sequencing Service 
- Single Cell Sequencing 
- Spatial transcriptome sequencing 
  - Spatial Gene Expression for Fresh Frozen
  - Spatial Gene Expression for FFPE
High throughput sequencing services 
Cell-based immune-related assays 
Proteomics testing service 
- Proteomics services by LC/MS 
- Olink Proteomics Services 
  - Olink Precision Proteomics Analysis Service
Small molecule metabolite detection services 
- Metabolomics service 
  - Targeted metabolomics
  - Non-targeted metabolomics
- Nucleic acid mass spectrometry detection service

Single Cell Immune Profiling

Product Description

Case Analysis

Result display

Sample delivery

Q&A

In 2015, 10x Genomics released the Chromium Single Cell System platform based on microfluidic and oil-drop wrapping technologies, enabling high-throughput single-cell transcriptome and single-cell V(D)J sequencing. This technology not only allows perfect matching of TCR/BCR duplexes to obtain TCR/BCR duplex composition, but also could be detailed into the single cell level while obtaining 5' end transcriptome information.

New released Single Cell Immunoprofiling of 10x Genomics uses microfluidic chip to prepare single cell systems, selecting universal primers for the 5' end-joins and nested primers for the immune molecular constancy region for V(D)J enrichment to enable full-length sequencing of paired heavy and light chains (B cells) or alpha and beta chains (T cells) at the single cell level. Full-length sequencing is performed, thus providing an efficient technical platform for comprehensive and systematic studies of immunomic libraries, and is important for the study of the molecular mechanisms of disease onset and development of disease.

Sample types

Animal tissues, other body fluids such as blood, single cell suspensions, cell lines, etc.

Experimental process

Experimental flow of 10x Genomics single cell immune group library

Analysis process

Flow Chart of Bioinformatics Analysis of Single Cell VDJ Immunogroup Bank

Application directions

a. Identification of antibodies and discovery of new antibodies;

b. Revealing the development of Ig libraries and mapping global Ig libraries;

c. Studying infectious diseases, such as the evolutionary spectrum of antibodies in the development of infections;

d. Studying the propensity for V(D)J recombination in immune disorders and to search for biomarkers of immune disorders;

e. Early detection of cancer, and studying markers for the diagnosis of cancer progression and recurrence, and studying tumor immune mechanisms.

Product advantages

a. Identification and characterization of rare cell types and biomarkers;

b. Analysis of tissue micro environment, disease progression and drug immune response;

c. Large-scale antibody and TCR discovery against neoantigens

d. Characterizing the immune response to infection by measuring immune cell phenotypes;

e. Reduce single cell sequencing costs through large-scale single-cell V(D)J+ expression profiling sequencing;

f. Provide flow cytometry sorting services to enable sorting for immune cells with purposes.

Case 1 Mapping CD8+ T cells in the gut of patients with ulcerative colitis by single-cell sequencing

Lymphocytes in the colon, such as tissue-resident memory CD8+ T cells, can respond rapidly to repeated antigen exposure. However, their cellular phenotype and the mechanisms by which they drive immune regulation and inflammation remain unclear. Using single-cell immunome library sequencing and mass cytometry, the researchers mapped the cellular profile of CD8+ T cells in healthy colon and ulcerative colitis (UC). The study found broad heterogeneity in CD8+ T cell composition, including expanding effector cells and post-terminally differentiated effector CD8+ T cells. UC-associated CD8+ effector T cells can trigger tissue destruction and produce the tumor necrosis factor TNF-α, while post-terminally differentiated effector CD8+ T cells have access to innate signals to perform regulatory functions that may alleviate excessive inflammation. The study identifies the colonic CD8+ T cell phenotype in healthy control and UC patients, make clear their clonal relationship and describes dysfunctional terminally differentiated CD8+ T cells expressing IL-26 in UC, which could attenuate symptoms of acute colitis in an IL-26 transgenic mouse model.

Reference：Corridoni D, Antanaviciute A, Gupta T, et al. Single-cell atlas of colonic CD8+ T cells in ulcerative colitis[J]. Nat Med,2020,91480-1490.

Case 2 Molecular pathways of colonic inflammation induced by tumor immunotherapy

Checkpoint blockade using specific antibodies against PD-1 and CTLA-4 inhibitory receptors induces durable responses in a variety of human cancers, but can also induce severe inflammatory toxicity. Nearly 60% of patients treated with the combination of PD-1 and CTLA-4 antibodies experience severe treatment-limiting toxicity, and the cellular and molecular mechanisms of these immune-related toxicities are unclear, which delays the optimal timing of clinical treatment.

In this paper, the researchers used 10x immunome library sequencing and flow cytometry analysis of intestinal samples from tumor patients after immunotherapy to perform a comprehensive analysis of immune cells in colitis. The results showed a significant accumulation of CD8+ T cells in a highly cytotoxic and proliferative state, but no evidence was found from regulatory T cell depletion. T cell receptor (TCR) sequence analysis showed that the majority of CD8+ T cells associated with colitis were from tissue-resident populations, explaining the early onset of colitis symptoms after treatment initiation. The analysis also identified cytokines, chemokines and surface receptors which could be served as therapeutic targets for colitis and other potential inflammatory side effects of checkpoint blockade.

Reference：Luoma AM,Suo S,Williams HL,et al.Molecular Pathways of Colon Inflammation Induced by Cancer Immunotherapy[J].Cell,2020,3:655-671.e22.

Case 3 Identification of a potent neutralizing antibody against SARS-CoV-2 by high-throughput single-cell sequencing of B cells from recovering patients

COVID-19 pandemic urgently requires therapeutic and preventive interventions urgently. This study reports the rapid identification of SARS-CoV-2 neutralizing antibodies by high-throughput single-cell RNA and V(D)J sequencing of antigen-rich B cells from 60 recovering patients. From 8,558 antigen-binding IgG1+ clonotypes, 14 potent neutralising antibodies were identified, the most potent of which was BD-368-2 with IC50 of 1.2 and 15 ng/mL against pseudo- and true SARS-CoV-2, respectively. BD-368-2 also showed potent therapeutic and prophylactic efficacy. In addition, the 3.8 Å cryo-electron microscopic structure of the neutralizing antibody in complex with a spike-extracellular domain trimer revealed overlapping epitope ACE2 binding sites for the antibody. Based on the predictions, it was demonstrated that the SARS-CoV-2 neutralizing antibody has a similar CDR3H structure to the SARS-CoV neutralizing antibody. It was shown that human neutralizing antibodies can be effectively identified by high-throughput B-cell sequencing in response to epidemic infectious diseases.

Reference：Cao Y, Su B, Guo X , et al. Potent neutralizing antibodies against SARS-CoV-2 identified by high-throughput single-cell sequencing of convalescent Patients' B cells[J]. Cell, 2020.