In the field of next-generation probiotics (NGPs), understanding the immunomodulatory effects of microbial strains is essential for defining their mechanisms of action (MOAs). Probiotic-host interactions are often mediated through complex immunological pathways, including cytokine regulation, T cell differentiation, dendritic cell maturation, and modulation of epithelial immune responses. These immunomodulatory characteristics not only shape the physiological outcome of host-microbe interactions but also determine the strain's potential utility in areas such as metabolic regulation, mucosal immunity, and microbiome stability.
At Creative Biolabs, we offer a dedicated platform for immunomodulation assays tailored to elucidate the immune-related MOAs of probiotics. This service supports clients aiming to select, characterize, and optimize probiotic candidates for downstream development of live biotherapeutic products (LBPs).
Importance of Immunomodulatory Profiling in Probiotic Research
As regulatory agencies increasingly emphasize the importance of mechanistic insights for live biotherapeutic development, functional assays that capture strain-specific immunomodulatory properties are indispensable. These assays provide crucial data for GRAS status, IND filings, and microbiome-targeted product claims. Understanding the immunomodulatory profile of a probiotic strain also helps differentiate it in a competitive market saturated with generic formulations.
Moreover, recent advances in systems immunology and microbiome science highlight the centrality of immune modulation in mediating probiotic effects—be it through increasing Treg populations, modulating dendritic cell responses, or attenuating pro-inflammatory cytokine cascades. Creative Biolabs' Immunomodulation Assays bridge this scientific need by offering multi-dimensional tools for immune-function discovery.
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Custom Immunomodulation Assay Platform
Creative Biolabs has developed an integrated immunological testing platform designed to interrogate both innate and adaptive immune responses triggered by probiotic strains. This platform leverages a broad array of validated cellular models, multiplexed detection technologies, and pathway-specific functional assays to dissect the immunomodulatory capacity of next-generation probiotics.
Key assay capabilities include:
Cytokine Profiling via ELISA, Luminex, and MSD
Quantify cytokine and chemokine signatures in cell supernatants post-stimulation with probiotic samples. Targeted panels include pro-inflammatory (e.g., IL-6, TNF-α), anti-inflammatory (e.g., IL-10, TGF-β), and mucosal immunity-related markers (e.g., IL-22, IL-17A).
T Cell Polarization Assays
Analyze how probiotics influence differentiation of naïve CD4+ T cells into Th1, Th2, Th17, or Treg lineages using intracellular cytokine staining and transcription factor profiling (e.g., T-bet, GATA3, RORγt, FOXP3). Both primary human PBMC and murine splenocyte models are available.
Dendritic Cell (DC) Maturation and Activation
Evaluate upregulation of costimulatory molecules (CD80, CD86, CD40), MHC class II expression, and cytokine secretion in monocyte-derived DCs upon probiotic stimulation, offering insights into antigen-presenting potential and T cell activation pathways.
Macrophage Polarization Analysis (M1 vs. M2)
Utilize THP-1-derived macrophages or murine BMDMs to assess expression of M1 markers (iNOS, IL-1β, IL-6) versus M2 markers (Arg1, CD206, IL-10), coupled with metabolic readouts (e.g., glycolysis vs. oxidative phosphorylation shifts) to characterize immunometabolic
NF-κB and MAPK Pathway Activation
Employ HEK293 or RAW264.7 reporter cell lines engineered with NF-κB or AP-1 luciferase constructs, alongside phospho-protein detection assays (e.g., p-p65, p-ERK, p-JNK), to map probiotic-driven signal transduction events.
Epithelial Cell–Immune Cell Crosstalk Models
Simulate gut barrier conditions using Caco-2 or HT-29 epithelial monolayers co-cultured with PBMCs, THP-1, or DCs across Transwell systems. These models allow for compartmentalized analysis of epithelial cytokine responses, tight junction integrity (via TEER), and immune cell modulation.
Conditioned Media and Postbiotic Analysis
In addition to whole-cell probiotics, our platform supports the evaluation of probiotic supernatants, heat-killed cells, or isolated metabolites (postbiotics) for their immunological effects, enabling non-viable candidate screening under regulatory-compliant frameworks.
All assay formats are available in 96- or 384-well high-throughput configurations for screening purposes or can be scaled to customized formats for in-depth mechanistic studies. Strain combinations, matrix-specific testing (e.g., dairy, capsule, lyophilized powders), and time-course kinetics are also supported. This modular and highly adaptable system ensures Creative Biolabs can accommodate a wide range of client-specific objectives—from early-stage strain scouting to preclinical candidate validation.
Streamlined Assay Workflow for Immune MOA Evaluation
Our optimized workflow ensures reproducibility, scalability, and scientific rigor across all immunomodulation assays. The standard pipeline is as follows:
Comprehensive Deliverables for Mechanistic Clarity
Clients working with Creative Biolabs can expect an end-to-end analytical package that enables strategic decision-making and regulatory compliance. Deliverables include:
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Standardized and validated immunomodulation assay reports
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Quantitative immune response profiles
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Cytokine secretion charts and heatmaps
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Immune cell phenotyping data (flow cytometry plots, polarization ratios)
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Pathway modulation analysis
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Recommendations for lead strain selection and downstream testing
All data packages are delivered in both raw and processed formats, with full traceability of biological materials and SOPs used.
Where Immunomodulation Assays Make an Impact
Live Biotherapeutic Product Development
Mechanistic validation of probiotic strains is essential for advancing LBPs through regulatory pipelines. Immunomodulation assays support strain selection, optimization, and GRAS/IND documentation by elucidating strain-specific effects on host immunity.
Gut-Immune Axis Studies
Understanding how specific probiotics influence gut-associated lymphoid tissue (GALT), dendritic cell trafficking, or cytokine gradients in the lamina propria is crucial for research into immune tolerance and mucosal immunity.
Metabolic Disorder Research
Several probiotic strains modulate systemic inflammation—a key driver of insulin resistance and obesity. Creative Biolabs' immune assays reveal anti-inflammatory mechanisms through modulation of TNF-α, IL-1β, and IL-10 profiles.
Personalized Microbiome Interventions
As personalized microbiome interventions gain traction, immune profiling can help match probiotic strains with individual immune phenotypes. This supports precision nutrition and microbiome-guided health solutions.
Veterinary and Animal Health R&D
Our platform is fully adaptable for veterinary probiotics. Immunomodulatory screening aids the selection of microbial candidates capable of enhancing livestock or companion animal immune health.
Explore Related Functional Screening Solutions
Creative Biolabs offers an extended suite of MOA discovery services to complement immunomodulation analysis:
Creative Biolabs remains committed to enabling mechanistic clarity for microbiome innovators worldwide. With advanced immunology infrastructure, expert-led assay design, and customizable analytics, our Immunomodulation Assay platform offers the precision and scalability required for modern probiotic R&D. To request a custom project proposal or consultation, please contact us today.
FAQs
What is the mechanism of action of probiotics on immune cells?
Probiotics interact with immune cells through pattern recognition receptors, modulate cytokine secretion, influence T cell differentiation, and alter dendritic and macrophage activation to shape immune homeostasis and mucosal tolerance.
How do immunomodulation assays help in selecting probiotic lead strains?
These assays quantify cytokine profiles, immune cell polarization, and signaling pathway activation, helping identify strains with desirable immunological effects for advancing into live biotherapeutic development.
Can immunomodulation assays be integrated with other MOA screening services?
Absolutely. These assays are part of our broader MOA discovery platform and can be combined with stress response profiling, metabolic assays, or host-microbe interaction tests for comprehensive evaluation.
Other Resources
References
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Bron, Peter A., et al. "Can probiotics modulate human disease by impacting intestinal barrier function?." British Journal of Nutrition 117.1 (2017): 93-107. https://doi.org/10.1017/S0007114516004037
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Cristofori, Fernanda, et al. "Anti-inflammatory and immunomodulatory effects of probiotics in gut inflammation: a door to the body." Frontiers in immunology 12 (2021): 578386. https://doi.org/10.3389/fimmu.2021.578386
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Mohamadzadeh, Mansour, et al. "Lactobacilli activate human dendritic cells that skew T cells toward T helper 1 polarization." Proceedings of the National Academy of Sciences 102.8 (2005): 2880-2885. https://doi.org/10.1073/pnas.0500098102
