Cytokine Storm Testing

Immune Monitoring Solutions for COVID-19 Research

Cytokine Storm Testing for SARS-CoV-2 & COVID-19 Research

Rules-Based Medicine (RBM) is a CLIA certified laboratory providing researchers with reproducible, quantitative immunoassay data from a single drop of blood serum or plasma.

We develop and manufacture our own assays in order to control for lot-to-lot variability and minimize matrix interferences (RF factor, heterophile antibodies).

InflammationMAP® and Immune Markers for COVID-19

Measure 55 key inflammation markers in a cost-effective, and efficient manner from 5 validated multiplex panels covering inflammatory pathways including cytokines, chemokines, vascular proteins and acute-phase reactants. Our biomarker testing services have been used in hundreds of clinical trials and research projects and cited in over 1000 peer reviewed publications.

Blue Text = Multiplex Immunoassays | Ultrasensitive Immunoassay = Ultrasensitive Immunoassays

InflammationMAP®

CORE1 (CytokineMAP®A)
1. GM-CSF 2, 13, 19
2. IFN-gamma 3, 4, 5, 19, 20
3. IL-2 12, 16, 19
4. IL-3
5. IL-4 19
6. IL-5
7. IL-6 10, 11, 13, 14, 19
8. IL-7 16, 19
9. IL-8 1, 13, 19
10. IL-10 1, 11, 13, 19
11. IL-18
12. MIP-1 alpha 16, 19
13. MIP-1 beta 19
14. MCP-1 1, 13, 18
15. TNF-alpha 13, 15, 16, 19
16. TNF-beta 13, 15
Core2 (CytokineMAP®B)
1. BDNF
2. Eotaxin-1
3. Factor VII
4. ICAM-1
5. IL-1 alpha
6. IL-1 beta 15, 18, 19
7. IL-1ra1
8. IL-12p40
9. IL-12p70
10. IL-17 19
11. IL-23
12. MMP-3 18
13. MMP-9
14. SCF1
15. VEGF 19
HMP8
1. Adiponectin
2. A2Macro
3. EN-RAGE1
4. Ferritin 16
5. Myoglobin1
6. PAI-1
7. PARC
8. RANTES 20
9. TIMP-11
10. TNFR2
11. VCAM-1
HMP2
1. Lp(a)
2. B2M
3. CRP 13
4. SAP1
5. TBG
6. vWF1
Core4
1. AAT1
2. C3
3. Fibrinogen
4. Haptoglobin
5. Ig A 17
6. Ig M 17
7. VDBP

Additional Markers

1. Interferon alpha 3, 4,5
2. Interferon beta 3, 4, 5  Ultrasensitive Immunoassay
3. LIGHT 6, 13  Ultrasensitive Immunoassay
4. Neuropilin-1 7, 8
5. Growth/differentiation factor 15 9
6. Interferon gamma Induced Protein 10 11, 14, 19
7. Interleukin-2 receptor alpha 12
8. LAP-TGF-beta 20
9. Carbonic anhydrase 9 1
10. Decorin 1

References

  1. Distinct patterns of blood cytokines beyond a cytokine storm predict mortality in COVID-19
    (2021) Herr C, et al. Journal of Inflammation Research.
    https://doi.org/10.2147/JIR.S320685
  2. GM-CSF-based treatments in COVID-19: reconciling opposing therapeutic approaches
    (2020) Lang FM, et al. Nature Reviews Immunology. 20, 507–514 (2020)
    https://doi.org/10.1038/s41577-020-0357-7
  3. Impaired type I interferon activity and inflammatory responses in severe COVID-19 patients.
    Hadjadj J, et al. Science 369(6504):718-724.v
    Recent coverage of the Hadjadj J, et al. article in BioSpace
  4. Type III interferons disrupt the lung epithelial barrier upon viral recognition.
    (2020) Broggi A, et al. Science 369(6504):706-712
  5. Type I and III interferons disrupt lung epithelial repair during recovery from viral infection
     Major J, et al. Science 369(6504):712-717
  6. Cerecor and Myriad Genetics announce that levels of novel cytokine, LIGHT were highly correlated with disease severity and mortality in COVID-19 ARDS biomarker study
    Harrell J and Gleason S. (2020) https://myriadrbm.com/2020/05/26/cerecor-and-myriad-genetics-announce-that-levels-of-novel-cytokine-light-were-highly-correlated-with-disease-severity-and-mortality-in-covid-19-ards-biomarker-study/
  7. Neuropilin-1 is a host factor for SARS-CoV2 infection
    (2020) Daly JL, et al. Science.
    DOI:10.1126/science.abd307
  8. Neuropilin-1 facilitates SARS-CoV2 cell entry and infectivity. (2020) Cantuti-Castelvetri L, et al. Science
    DOI: 10.1126/science.abd2985
  9. Growth differentiation factor-15 provides prognostic information superior to established cardiovascular and inflammatory biomarkers in unselected patients hospitalized with COVID-19
    (2020) Myhre PL, et al. Circulation 
    DOI: 10.1161/CIRCULATIONAHA.120.050360
  10. The role of Interleukin 6 inhibitors in therapy of severe COVID-19
    (2020) Nasanov E, et al. Biomedicine & Pharmacotherapy Volume 131, 2020
    https://doi.org/10.1016/j.biopha.2020.110698
  11. A dynamic COVID-19 immune signature includes associations with poor prognosis
    (2020) Laing AG, et al. Nature Medicine. 26, 1623–1635 (2020)
    https://doi.org/10.1038/s41591-020-1038-6
  12. The Inhibition of IL-2/IL-2R gives rise to CD8+T cell and lymphocyte decrease through JAK1-STAT5 in critical patients with COVID-19 pneumonia
    (2020) Shi H, et al. Cell Death Dis. 2020 Jun 8;11(6):429.
    DOI: 10.1038/s41419-020-2636-4
  13. Biomarkers of COVID-19 and technologies to combat SARS-CoV2
    (2020) Zhang L and Guo H. Advances in Biomarker Sciences and Technology. 2020;2:1-23
    doi: 10.1016/j.abst.2020.08.0011,
  14. Longitudinal immune profiling reveals key myeloid signatures associated with COVID-19
    (2020) Mann ER, et al. Science Immunology 5,5. eabd6197
    DOI:10.1126/sciimmunol.abd6197
  15. Immunophenotyping of COVID-19 and influenza highlights the role of type I interferons in development of severe COVID-19
    (2020) Lee LS, et al. Science Immunology. 5, 49, eabd1554
    DOI:10.1126/sciimmunol.abd1554
  16. COVID-19: consider cytokine syndromes and immunosuppression
    (2020) Mehta P, et al. The Lancet. 395, 10229. 1033-1034
    https://doi.org/10.1016/S0140-6736(20)30628-0
  17. Serum IgA, IgM, and IgG response in COVID-19
    (2020) Ma H, et al. Cellular & Molecular Immunology. 17, 773-775
    https://doi.org/10.1038/s41423-020-0474-z
  18. Matrix metalloproteinase 3 as a valuable marker for patients with COVID-19
    (2020) Shi S, et al. J Med Virol. 10.1002/jmv.26235
    DOI: 10.1002/jmv.26235
  19. Inhibition of cytokine signaling by ruxolitinib and implications for COVID-19 treatment
    (2020) Yeleswarm S, et al. Clinical Immunology. 218, 108517
    https://doi.org/10.1016/j.clim.2020.108517
  20. Interferon gamma, TGF-b1 and RANTES expression in upper airway samples from SARS-CoV2 infected patients
    (2020) Clinical Immunology. 220, 108576.
    https://doi.org/10.1016/j.clim.2020.108576

SARS-CoV-2 T cell activation with TruCulture
Spike Protein tubes

SARS-CoV-2 Spike Protein TruCulture Tubes (2019 nCoV)
SARS-CoV-2 Spike Protein TruCulture Tubes (Delta variant) NEW!

TruCulture is a whole blood collection and culture system for immune monitoring in subjects and has been utilized in hundreds of clinical trials. The SARS-CoV-2 Spike Protein TruCulture tube was designed and developed to be used in clinical trials to:

  1. Quantify SARS-CoV-2 vaccine-dependent T cell activation.
  2. Investigate the magnitude and duration of T cell responses post SARS-CoV-2 infection.

Use TruCulture tubes to collect 1 mL of whole blood directly from a patient to immediately start antigen recognition, processing, and presentation in the tube. Each TruCulture tube contains 2 mL of media including the SARS-CoV-2 spike protein antigen. After 48 hours at 37o C, the secreted T cell cytokines IFNγ and IL-2 are quantified by validated immunoassays (RBM’s ultrasensitive immunoassay services).

In comparison to ELISpot utilizing peptide pools, TruCulture recombinant protein antigen recall technology for clinical studies has many advantages:

  • More representative of cell mediated immunity with the entire immune system present for the complex interplay of antigen processing, presentation, and T cell recognition
  • More sensitive – a higher proportion of viable cells (cell reaction starts immediately upon blood draw)
  • Closed sterile system performed with a simple blood draw by a phlebotomist at a trial site with a 37°C degree dry-heat block being the only specialized equipment needed
  • More cost-effective
  • Utility recently published in Lancet Infectious Disease

Download the SARS-CoV-2 Spike Protein TruCulture Tubes application note to learn more.

NEW! SARS-CoV-2 Spike Protein TruCulture tube (Delta variant)

In comparison to the original Wuhan-1 SARS-CoV-2 strain, the Delta variant (B.1617.2) Spike protein is defined by upwards of 10 amino acid substitutions in the SARS-CoV-2 spike protein. Four of these substitutions are of concern and are thought to be the primary reason for higher degree of transmission and possibly causing more severe disease1:

  1. D614G -shared with other highly transmissible Variants of Concern (VOC) (α, β, γ)
  2. T478K -Receptor-Binding Domain (RBD) region
  3. L452R -RBD region, higher affinity for angiotensin-converting enzyme 2 (ACE2), more difficult for immune recognition
  4. P681R -enhances S precursor furin cleavage to active S1/S2 configuration

The remaining six Delta defining Spike protein amino acid changes are:

  1. G142D
  2. T19R
  3. R158G
  4. D950N
  5. E156del
  6. F157del

All ten substitutions and deletions will likely have an important impact on T cell epitope (Class I and II MHC) presentation and recognition by T cells.  For previously vaccinated or infected subjects, or new vaccine development studies, the SARS-CoV-2 Delta Variant Spike Protein TruCulture Tube is a powerful addition to the original TruCulture Spike protein tubes for quantifying antigen recall and cell mediated immunity.

Reference:

  1. https://outbreak.info/situation-reports#Lineage_Mutation

For other TruCulture Tubes and Stimulants, click here.

Analysis

TruCulture cell pellets can be collected for downstream analyses:

Application Note Download

Download the SARS-CoV-2 Spike Protein TruCulture Tubes application note to learn how TruCulture tubes are used to measure antigen recall responses to the SARS-CoV-2 spike protein.

TruCulture Procedure, Collect

01. COLLECT

Draw 1 mL of blood directly into the TruCulture Tube and break off the plunger.

TruCulture Procedure, MIX

02. MIX

Gently invert tube to mix 3 to 5 times

TruCulture Procedure, Incubate

03. INCUBATE

Place tube in 37ºC heat block for up to 24 or 48 hours.

TruCulture Procedure, separate

04. SEPARATE

Manually insert valve to separate supernatant from the cells. Collect supernatant and cell layer for downstream analysis.

Watch our instructional video to see how easy it is to use TruCulture.

All you need is a phlebotomist and a 37oC heat block!
No need for specialized cell culture equipment or technical knowledge.

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Multi-Analyte Profile (MAP) Platform

The RBM MAP platform has been built and optimized to help researchers discover and validate biomarker patterns for use in their drug and diagnostic development efforts. Robust multiplexing on the microsphere-based Luminex technology combined with automated liquid handling, delivers more data in less time than other platforms, resulting in comparable data to traditional ELISA assays.

  • High throughput multiplexing with automated liquid handling reduces sample volume requirements
  • Enhanced accuracy with use of multi-level controls, standard curves, and proprietary blockers
  • High quality and reproducibility with stringent quality control parameters for reliable data

We ensure quality for every multiplex, every analyte, every sample, every time.
The RBM platform produces precise and dependable results by utilizing automated liquid handling systems, advanced quality monitoring, validated data reporting processes, and a highly-trained and dedicated staff.

Every analyte included in RBM’s Multi-Analyte Profiles (MAPs) has its own set of calibrators and controls to provide reliable, high quality data. By continuous quality improvement, RBM ensures that customers are satisfied with the products and services they receive.

xMAP

Other RBM Products

CustomMAP

Our CustomMAP is a flexible option to let you pick specific biomarkers for your research needs. Customize the number of multiplexes to build your CustomMAP.

Speak to a sales representative to learn more about CustomMAP options.

50 sample minimum required. Volume requirements are dependent on the number of multiplexes selected.

Ultrasensitive Immunoassays - Simoa

Looking for trace quantities of markers? Ultrasensitive immunoassays achieve orders-of-magnitude greater sensitivity than conventional immunoassay platforms for when levels of key proteins are extremely low in serum or plasma.

simoa

SARS-CoV-2 Vaccine Development Application with TruCulture®

Utilized in hundreds of clinical trials for a variety of applications, TruCulture® is a whole blood collection and culture system for immune monitoring of subjects in clinical trials.

  • Detect vaccine-dependent T-cell activation (antigen recall)
  • Early safety studies of vaccines and their individual components
  • Profile a subject’s functional immune status
TruCulture Vaccine Development Application note cover page