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Creative Biolabs offers single cell cytokine profiling services to advance the proteomic innovations at the single cell level. With the cytokine profiling data, we can discover the functional cell phenotype of each cell, which enables customers to advance the future of medicines against diseases.

Single Cell Cytokine Profiling

Single Cell Cytokine Profiling is a comprehensive functional profiling of each cell type by using a single-cell chip and running an entire highly-multiplexed proteomics workflow. This technology can be used as a new standard in single-cell functional biology. What you need to do is sending your sample to us, and we will send back the fully analyzed data to you.

Our Single Cell Cytokine Profiling Services

"Proteomic Barcoded" Chip is able to analyze the "functional phenotype" of each immune cell (e.g. T cell, monocyte, and NK cell) by detecting 30+ cytokines per cell and provide researchers with more accurate data. To detect how immune cells proteomically perform against a heterogeneous system, we can use single-cell adaptive immune solution to measure the "functional phenotype" of each immune cell, thus allowing for complete single-cell functional characterization and revealing functional biological drivers of persistence, potency, durability etc. In addition, we can offer single-cell functional biology analysis for human, mouse and NHP samples.

Each immune cell is different, and we need a more comprehensive understanding of its cell characteristics. This will help us connect and differentiate with other immune cells.

For single-cells, cytoking profiling service can address the limitations of RNA seq and flow cytometry by using cytokines to define cell phenypes more completely.

Single Cell Cytokine Profiling workflow.Figure 1. Single Cell Cytokine Profiling workflow.

Here are the cytoking profiling panels for your selection.

  • Cytokine Panels
  • 1) Human Adaptive Immune

    Granzyme B, IFN-γ, MIP-1α, Perforin, TNF-α, TNF-β, GM-CSF, IL-2, IL-5, IL-7, IL-8, IL-9, IL-12, IL-15, IL-21, CCL11, IP-10, MIP-1β, RANTES, IL-4, IL-10, IL-13, IL-22, TGFβ1, sCD137, sCD40L, IL-1β, IL-6, IL-17A, IL-17F, MCP-1, MCP-4.

    2) Non-Human Primate Adaptive Immune

    TNF-α, MCP-1, IL-2, IL-4, MIP-1β, IL-6, IL-8, IL-1β, RANTES, IFN-g, IP-10, MIP-1α, MIF, GM-CSF.

    3) Mouse Adaptive Immune

    Granzyme B, IFN-γ, MIP-1α, TNF-α, GM-CSF, IL-2, IL-5, IL-7, IL-12p70, IL-15, IL-21, sCD137, CCL11, CXCL1, CXCL13, IP-10, RANTES, Fas, IL-4, IL-10, IL-13, IL-27, TGFβ1, IL-6, IL-17A, MCP-1, IL-1β.

    4) Human Natural Killer (Coming Soon)

    Granzyme B, IFN-γ, MIP-1α, Perforin, TNF-α, TNF-β, GM-CSF, IL-2, IL-5, IL-7, IL-8, IL-9, IL-12, IL-15, IL-21, CCL11, IP-10, MIP-1β, RANTES, IL-4, IL-10, IL-13, IL-22, TGFβ1, sCD137, sCD40L, IL-1β, IL-6, IL-17A, IL-17F, MCP-1, MCP-4.

    5) Human Innate Immune

    IFN-γ, MIP-1α, TNF-α, TNF-β, GM-CSF, IL-8, IL-9, IL-15, IL-18, TGF-α, IL-5, CCL11, IP-10, MIP-1β, RANTES, BCA-1, IL-10, IL-13, IL-22, sCD40L, IL-1β, IL-6, IL-12-p40, IL-12, IL-17A, IL-17F, MCP-1, MCP-4, MIF, EGF, PDGF-BB, VEGF.

    6) Human Inflammation

    GM-CSF, IFN-γ, IL-2, IL-12, TNF-α, TNF-β, IL-4, IL-5, IL-7, IL-9, IL-13, CCL11, IL-8, IP-10, MCP-1, MCP-4, MIP-1α, MIP-1β, RANTES, IL-10, IL-15, IL-22, TGF-β1, IL-1β, IL-6, IL-17A, IL-17F, IL-21, Granzyme B, Perforin, sCD40L, sCD137.

    7) Mouse Innate Immune

    IFN-g, TNF-a, MIP-1a, IL-15, GM-CSF, IL-5, IL-10, IL-13, IL-6, IL-17A, MCP-1, IP-10, MIP-1b, EGF, PDGF-BB, MIF.

  • Subset Panels
  • 1) Membrane Stain

Advantage of Single Cell Cytokine Profiling

  • 32+ Cytokine Analysis
  • Multiplexed 32+ cytokine analysis at single-cell resolution.

  • Functional Phenotyping
  • Functional Phenotyping targeting 500-1500 single cells per chip.

  • Single-cell input levels for high-quality samples
  • Fully automated, quick, and cost-efficient workflow.

Applications of Single Cell Cytokine Profiling

  • Application of single-cell functional proteomics for novel biomarker discovery

Functional immune profiling with single-cell proteomics can analyze changes in polyfunctional monocytes according to the severity of the disease. Therefore, accelerate the comprehensive understanding of the immune response can be gained based on potential biomarkers. This is essential for identifying and developing effective treatments.

Paper Title Multi-omics resolves a sharp disease-state shift between mild and moderate COVID-19
Technology Single-cell multiplex secretome assay and Single cell multi-omics assay
Journal Cell
IF 41.582
Published 2020
Abstract In the study of 139 COVID-19 patients, they represented all levels of disease severity. The major shift between mild and moderate disease was determined by clinical measurements, immune cells, single-cell multiplex secretome assay and single cell multi-omics assay. Elevated inflammatory signaling is accompanied by the loss of specific classes of metabolites and metabolic processes
Sample COVID-19 patients and healthy control samples
Methods 32 secreted cytokines in individual live CD8 + T cells were measured
Result Polyfunctionality, as quantified by the Polyfunctional Strength Index (PSI = the numbers of different proteins secreted × copy numbers secreted), is similar for healthy and mild cases, but up-regulated at moderate severity. Thus, polyfunctionality is reduced in severe patients

Single cell polyfunctional strength index (PSI) of CD8 + T cells according to sample WOS (Su et al., 2020).Figure 2. Single cell polyfunctional strength index (PSI) of CD8 + T cells according to sample WOS (Su et al., 2020).

  • Reveal sources of immune modulation with the single-cell innate immune solution

Single-Cell Innate Immunity solution is helping researchers detect functional cellular differences associated with immune regulation.

  • Understanding the role of innate and myeloid cells in immune suppression

In cancer immunology, myeloid cells (e.g. Neutrophils, MDSCs, and Monocytes) are key components of the immune system, largely responsible for innate defense against an array of pathogens (Figure 3). Furthermore, myeloid cells provide key aspects of both orchestrating the attack of and suppression of the immune system towards the tumor.

Currently, myeloid cell types are being tested using Single-Cell Innate Immune solution for diseases such as cancer and inflammation.

Myeloid cells provide key aspects of both orchestrating the attack of and suppression of the immune system towards the tumor, analyzed on the Single-Cell Innate Immune solution(Ransohoff & Cardona, 2010).Figure 3. Myeloid cells provide key aspects of both orchestrating the attack of and suppression of the immune system towards the tumor, analyzed on the Single-Cell Innate Immune solution (Ransohoff & Cardona, 2010).

  • How the innate immune solution reveals monocyte response to pathogenic ligands?
Paper Title Highly multiplexed profiling of single-cell effector functions reveals deep functional heterogeneity in response to pathogenic ligands
Journal Proceedings of the National Academy of Sciences of yhe United States of America
IF 11.205
Published 2015
Abstract Currently, it remains a challenge to collect highly multiplexed measurements of secreted proteins from single cells for comprehensive analysis of functional states. Herein, Combine spatial and spectral encoding with polydimethylsiloxane (PDMS) microchambers for codetection of 42 immune effector proteins secreted from single cells. Using this platform to profile differentiated macrophages stimulated with lipopolysaccharide (LPS), the ligand of Toll-like receptor 4 (TLR4), reveals previously unobserved deep functional heterogeneity and varying levels of pathogenic activation. Uniquely protein profiling on the same single cells before and after LPS stimulation identified a role for macrophage inhibitory factor (MIF) to potentiate the activation of LPS-induced cytokine production. This technology enables full-spectrum dissection of immune functional states in response to pathogenic or environmental stimulation, and opens opportunities to quantify deep functional heterogeneity for more comprehensive and accurate immune monitoring
Methods Single-Cell Multiplex Protein Secretion Assay
Result The MIF-positive pop-ulation in a LPS-stimulated sample is replenished from the cells originating in other subpopulations in the basal state, suggesting the existence of phenotypic homeostasis for MIF production among all functional cell subsets. Within the PFA population, LPS induced a larger subset of PFA cells to secrete IL-6 and IL-10 compared with PAM3. Poly (I:C) appears to be the least potent ligand, with minimal increase of PFA population, although this could be due to the fact that our an-tibody panel did not include a large number of antiviral effectors. However, we noticed a significant phenotypic shift of MIF-secreting cells upon stimulation with poly (I:C)

Functional heterogeneity and subpopulations of U937-derived macrophages in response to TLR ligands (Lu et al., 2015).Figure 4. Functional heterogeneity and subpopulations of U937-derived macrophages in response to TLR ligands. (Lu et al., 2015)

  • Single-Cell innate immune solution application

Single-Cell Innate Immune solution is helping researchers detect functional cellular differences that correlate to immune suppression. The Single-Cell Innate Immune solution does this by revealing the sources of cellular differences based on the ability to detect what each immune cell is truly secreted in a highly multiplexed manner.

Single-Cell Innate Immune Panel measures over 30 cytokines that orchestrate the immune response from each innate and myeloid cell, back to the cell itself.Figure 5. Single-Cell innate immune panel measures over 30 cytokines that orchestrate the immune response from each innate and myeloid cell, back to the cell itself.

To learn more about the single cell cytokine profiling service, please feel free to contact us to confirm how we can be involved in your project.

References

  1. Lu, Y., Xue, Q., Eisele, M. R., Sulistijo, E. S., Brower, K., Han, L., Fan, R. (2015). Highly multiplexed profiling of single-cell effector functions reveals deep functional heterogeneity in response to pathogenic ligands. Proc Natl Acad Sci U S A, 112(7), E607-615.
  2. Ransohoff, R. M., & Cardona, A. E. (2010). The myeloid cells of the central nervous system parenchyma. Nature, 468(7321), 253-262.
  3. Su, Y., Chen, D., Yuan, D., Lausted, C., Choi, J., Dai, C. L., Heath, J. R. (2020). Multi-Omics Resolves a Sharp Disease-State Shift between Mild and Moderate COVID-19. Cell, 183(6), 1479-1495 e1420.
! ! For Research Use Only. Not for diagnostic or therapeutic purposes.

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