Th1 · Th2 · Th17 · Treg

Secreted Cytokine Readout

MSD Immune Cell Profiling — Th1/Th2/Th17/Treg Cytokine Panel Services

Characterize T-cell polarization, immune function, and treatment-associated immunomodulation by quantifying the secreted cytokines that define each T-helper subset — using MSD Multiplex panels purpose-built for immune cell profiling across human, mouse, and NHP.

T cells do not carry ID cards — their functional identity is defined by the cytokines they secrete. IFN-γ marks Th1; IL-4, IL-5, IL-13 mark Th2; IL-17A, IL-22 mark Th17; IL-10, TGF-β1 mark Treg. Flow cytometry identifies surface markers and intracellular cytokines, but MSD multiplex panels quantify the actual secreted effectors — the proteins that mediate immune function — from serum, plasma, cell culture supernatant, or CSF. The two approaches are complementary, not competing: flow cytometry tells you which cells are present; MSD tells you what those cells are producing. For vaccine research, immunotherapy monitoring, CAR-T cell characterization, and autoimmune disease profiling, the secreted cytokine profile is the functional readout that matters.

Inquire About Immune Profiling T-Cell Subset Panels

T-Helper Cell Subsets and Cytokine Signatures

Since the seminal Th1/Th2 paradigm was described by Mosmann and Coffman in 1986, the T-helper universe has expanded to include Th17, Treg, Tfh, Th9, and Th22 subsets — each defined by a characteristic cytokine secretion profile. These subsets are not binary states but continuous spectra, and immune responses typically involve coordinated shifts across multiple subsets simultaneously. Measuring one or two representative cytokines provides a fragmentary view; comprehensive immune profiling requires simultaneous quantification of the full panel of subset-defining cytokines.

 T-Helper Lineage Cytokine Profiles
Th1 — Cell-mediated immunity: IFN-γ (signature), TNF-α, IL-2. Drives macrophage activation, CD8+ T-cell responses, and intracellular pathogen clearance. Dominant in autoimmune conditions (RA, MS, Type 1 diabetes) and anti-tumor immunity.
Th2 — Humoral & Type 2 immunity: IL-4 (signature), IL-5, IL-13. Drives B-cell class switching to IgE, eosinophil activation, and mast cell responses. Dominant in allergy, asthma, atopic dermatitis, and helminth infection.
Th17 — Barrier & fungal immunity: IL-17A (signature), IL-17F, IL-21, IL-22, GM-CSF. Protects mucosal surfaces; pathologically drives psoriasis, ankylosing spondylitis, IBD, and MS. IL-23 is the key upstream polarizing cytokine; IL-12/IL-23p40 is the shared subunit.
Treg — Immune regulation: IL-10 (signature, shared), TGF-β1. Suppresses effector T-cell responses; maintains peripheral tolerance. Treg dysfunction implicated in autoimmunity; Treg excess can suppress anti-tumor immunity.
Tfh — B-cell help: IL-21 (signature). Drives germinal center formation and affinity maturation. Relevant to vaccine responses and antibody-mediated autoimmunity.
Th9 — Type 2 variant: IL-9 (signature). Amplifies mast cell and eosinophil responses. Implicated in allergic inflammation and tumor immunity.

Mosmann TR, Coffman RL (1986). Journal of Immunology. 136(7):2348–2357. PMID: 2419433 — The foundational paper establishing the Th1/Th2 paradigm.

T-Cell Subset Profiling Panels — Human, Mouse, and NHP

Pre-configured V-PLEX panels mapping to each T-helper lineage, with U-PLEX custom options for study-specific configurations.

Panel	Plex	Key Analytes	T-Cell Coverage	Species
Human Th1/Th2 10-Plex	10	IFN-γ, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13, TNF-α, GM-CSF	Th1 + Th2 + inflammatory	Human
Human Th17 7-Plex	7	IL-17A, IL-21, IL-22, IL-23, IL-27, IL-31, MIP-3α	Th17 + Tfh + regulatory	Human
Human Th17 9-Plex	9	IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-31, GM-CSF, TNF-α	Th17 + inflammatory context	Human
Human Th17 10-Plex	10	Extended: IL-17A, IL-17F, IL-17A/F, IL-21, IL-22, IL-23, IL-27, IL-31, GM-CSF, TNF-α	Th17 complete + IL-17 family	Human
Human T-Cell 14-Plex	14	GM-CSF, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-22, IL-23, TNF-α	Th1 + Th2 + Th17 + Tfh	Human
Mouse Th1/Th2 10-Plex	10	IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, KC/GRO, TNF-α, IL-1β	Th1 + Th2 + inflammatory	Mouse
Mouse Th17 9-Plex	9	IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-31, GM-CSF, TNF-α	Th17 complete	Mouse
Mouse T-Cell 14-Plex	14	GM-CSF, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-22, IL-23, TNF-α	Full T-helper spectrum	Mouse
NHP T-Cell 12-Plex	12	GM-CSF, IFN-γ, IL-2, IL-4, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-22, MIP-3α, TNF-α	Cross-species translational	NHP
NHP Th1/Th2 9-Plex	9	IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12/IL-23p40, IL-13, TNF-α	Th1 + Th2 core	NHP

Panel availability confirmed during project consultation. Custom U-PLEX panels can combine T-cell cytokines with additional immune markers (chemokines, checkpoint proteins, activation markers) for study-specific configurations.

MSD Multiplex vs. Flow Cytometry — Complementary, Not Competing

These are the two dominant technologies for immune profiling — each answers different questions. Understanding the complementarity is key to designing an effective immune monitoring strategy.

Dimension	MSD Multiplex	Flow Cytometry (ICS/Flow)
What It Measures	Secreted cytokines in biological fluids (serum, plasma, supernatant, CSF)	Intracellular cytokines + surface markers at single-cell resolution
Question Answered	"What is the functional cytokine output of the immune system?"	"Which specific cell types are producing which cytokines?"
Sample Type	Serum, plasma, CSF, cell culture supernatant — cell-free	Whole blood, PBMCs, tissue digests — requires viable cells
Throughput	96-well / 70 sec read; high-throughput cohort-compatible	Thousands of events per sample; lower sample throughput
Multiplex	Up to 54 analytes per 25 µL	30+ parameters with spectral flow cytometry
Data Type	Continuous concentration values (pg/mL)	Percentage of positive cells, MFI
Longitudinal Stability	Excellent — frozen samples, lot-bridged reagents	Requires fresh cells or cryopreserved PBMCs; batch effects possible
Best Use Case	Cohort studies, treatment monitoring, secreted cytokine profiling	Immune phenotyping, rare population identification, activation state

The most comprehensive immune monitoring programs use both: flow cytometry for cellular phenotyping (which cells are present and activated) and MSD multiplex for functional cytokine profiling (what those cells are secreting into circulation). Data from both platforms can be integrated to link cellular phenotypes with their secreted effector molecules.

Innate Immune Cell Profiling — Dendritic Cells & Macrophages

Extending immune profiling beyond the adaptive (T/B-cell) compartment to the innate immune cells that initiate and shape the immune response.

Panel	Plex	Analytes	Species	Application
Mouse DC Cell 5-Plex	5	IL-7, CTACK/CCL27, Fractalkine/CX3CL1, GRO-α/CXCL1, IL-12/IL-23p40, IP-10/CXCL10, MCP-1/CCL2, MIP-1β/CCL4, MIP-3α/CCL20	Mouse	Dendritic cell biology; antigen presentation
Mouse Macrophage 5-Plex	5	IL-10, IL-13, IL-4, MDC/CCL22, TARC/CCL17	Mouse	M1/M2 macrophage polarization
Mouse Macrophage 9-Plex	9	IL-15, IL-23, IL-6, TNF-α, IL-1β, IL-12p70, IP-10/CXCL10, MCP-1/CCL2, MIP-1α/CCL3	Mouse	Inflammatory macrophage activation; inflammasome research

Dendritic cell and macrophage panels complement T-cell profiling by capturing the innate immune signals that determine whether the adaptive response is tolerogenic or immunogenic. The macrophage panels distinguish M1 (pro-inflammatory: IL-6, TNF-α, IL-1β, IL-12p70) from M2 (anti-inflammatory: IL-10, IL-4, IL-13) polarization states. For comprehensive immune profiling — innate + adaptive — combine these with Th1/Th2/Th17 panels from the same sample. All services are for research use only.

CAR-T Cell Therapy Immune Monitoring

Specialized panels for CAR-T cytokine release, persistence, and immune effector function monitoring in research applications.

Cytokine Release Syndrome (CRS) Markers

IL-6, TNF-α, IFN-γ, IL-1β, IL-2, IL-8, IL-10, GM-CSF — the core cytokines elevated during CAR-T-associated CRS. Multiplex quantification from serial plasma samples enables kinetic profiling of CRS onset, peak, and resolution in preclinical and clinical research.

CAR-T Persistence & Effector Function

IL-7, IL-15 (T-cell survival/homeostasis), IFN-γ, TNF-α, IL-2 (effector function), IL-21 (Tfh/memory), GM-CSF (myeloid activation). Longitudinal profiling tracks CAR-T functional persistence and effector output over weeks to months post-infusion.

Immune Effector Cell-Associated Neurotoxicity (ICANS)

IL-6, IL-1β, TNF-α, IFN-γ, IL-8, MCP-1, IP-10 — cytokines and chemokines implicated in ICANS pathophysiology. CSF-plasma paired analysis can evaluate compartment-specific cytokine gradients across the blood-brain barrier during neurotoxicity.

CAR-T panels are available as pre-configured V-PLEX or customized via U-PLEX. All services are for research use only — not for clinical use.

Immune Cell Profiling Service Details

Service Package

Item	Full Service	Kit Only
Sample Processing	Included	— Self-run
Multi-Analyte Standard Curve	8-point, all panel analytes	Calibrators provided
QC Samples	3 levels, duplicate	— User-prepared
T-Cell Subset Analysis	Th1/Th2/Th17 indices	— Self-analysis
Raw Data	Included	— Instrument-dependent

For Research Use Only. Not for diagnostic procedures.

How to Get Started

Define Question

Are you profiling T-cell polarization, monitoring treatment immunomodulation, or characterizing CAR-T effector function? We'll match the panel to your immunological question.

Ship Samples

Serum, plasma, or cell culture supernatant on dry ice. 25 µL per well. Pre-analytical guidance for immune-functional readouts provided.

Interpret Immune Profile

Receive your quantitative cytokine data with Th1/Th2/Th17 subset indices and biological context for immune functional interpretation.

Immune Cell Profiling FAQ

How does MSD cytokine profiling complement flow cytometry for immune monitoring?

Flow cytometry identifies which cell types are present and whether they express activation markers or intracellular cytokines — it answers "which cells?" MSD multiplex measures the actual secreted cytokine concentrations in biological fluids — it answers "how much functional output?" The two are complementary: flow cytometry tells you that Th17 cells are expanded in a patient sample; MSD tells you whether those cells are secreting pathogenic levels of IL-17A. For comprehensive immune monitoring, the most informative programs use both: flow cytometry for cellular phenotyping and MSD for quantitative functional cytokine profiling. The MSD advantage for secreted cytokines is analytical: no permeabilization artifacts, no PMA/ionomycin stimulation bias, and direct quantification in pg/mL from the biological matrix of interest.

Which T-cell panel should I choose — Th1/Th2, Th17, or the T-Cell 14-Plex?

The choice depends on your immunological question. Th1/Th2 10-Plex covers the classical Type 1/Type 2 balance — ideal for vaccine response profiling, allergy/asthma studies, and basic immune phenotyping. Th17 panels (7/9/10-Plex) add the Th17/Tfh/regulatory axis — essential for autoimmune disease research, mucosal immunology, and CAR-T monitoring. T-Cell 14-Plex combines Th1, Th2, and Th17 markers in a single panel — best for comprehensive immune profiling where the T-cell polarization state is unknown or expected to shift across multiple subsets. Our scientific team can help match the panel to your specific research context during study design consultation.

Can I use these panels with cell culture supernatants from in vitro T-cell polarization assays?

Yes. MSD panels are compatible with cell culture supernatant from T-cell polarization, activation, suppression, and mixed lymphocyte reaction (MLR) assays. Key considerations: (1) serum-containing media introduces bovine cytokines — use low-serum or serum-free conditions where feasible, or include media-only controls to account for background; (2) the expected cytokine concentrations in culture supernatant are typically 10–1,000× higher than in plasma — MSD's wide dynamic range accommodates this without dilution in most cases; (3) for assays using PMA/ionomycin stimulation, note that stimulation conditions can affect the relative ratios of cytokines produced, and unstimulated controls are essential for interpretation. Contact us for cell culture-specific guidance during study design.

Do you offer species-matched panels for preclinical-to-clinical immune translation?

Yes. We offer T-cell profiling panels for human, mouse, and non-human primate (NHP) on the same MSD platform. This is a key advantage for translational immunology: the same detection technology, same sample volume (25 µL), same plate format, and same data output structure across species — enabling direct cross-species cytokine comparison without platform-switching variability. Mouse panels include the mouse-specific chemokine KC/GRO (functional homolog of human IL-8). NHP panels are validated for cynomolgus and rhesus macaque. Custom cross-species U-PLEX panels can also be configured for analytes shared across your species of interest.

Are these immune profiling panels validated for clinical use?

No. All T-cell profiling panels are manufacturer-validated for research use only (RUO). They are suitable for preclinical studies, cohort research, biomarker discovery, vaccine immunogenicity research, and CAR-T cell therapy investigational studies. They are not validated for clinical diagnostic procedures, patient management decisions, or as standalone clinical monitoring tools. For research programs that may later inform regulated work, we recommend discussing documentation requirements with our team during study design so we can align our processes with your downstream needs. All services are provided under a research-use-only framework.

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