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Decoding immune complexity

Scientific discoveries in the fields of immunology and molecular biology, have paved the way to the development of several new targeted therapies.

In this context, immunomonitoring data are essential to precisely determine the immune response of patients receiving these investigational drugs, and ultimately contribute to transitioning them from preclinical and clinical studies to new standards of care.

These data are even more critical when establishing the efficacy and safety of an immunomodulatory compound, because in this case, the establishment of a correlation between the immunological mechanism (the activation or inhibition of immune effectors) and the targeted clinical response (tumor regression, reduced viral load) conditions the further development of the product.

Evaluate overtime the number, the subtypes and the functionality of immune cells, the concentration and the activity of immune mediators is a complex scientific and technological challenge. To decode immune complexity, ABL has developed a unique expertise covering all facets of the immune response (innate and adaptive immunity) and all scales of analysis (cellular, biochemical and molecular).

ABL's team brings over 60 years of cumulated research in the field of immunology (see our publications), and continues strengthening its scientific expertise thanks to its frequent interactions with world-class immunologists and clinicians.

 

Using a large panel of technologies such as multi-parametric flow cytometry, Luminex, Singulex, Quanterix, ELISpot or ELISA, ABL offers a wide range of immunomonitoring solutions:


Flow cytometry

Flow cytometry is a powerful and versatile technique, ideal for phenotyping, enumerating and assessing the function of various immune cells. It allows simultaneous analysis of various parameters at the single cell level. Combined assessment of surface markers and intracellular antigens helps analyze the function of rare and precisely defined cell subsets. Functions assessed by flow cytometry include cytokine production, natural cytotoxicity, antibody dependent cell cytotoxicity (ADCC), and proliferation.

Flow cytometry can be performed on many types of cell suspension. ABL’s team has extensive experience in analyzing whole blood, primary cells (PBMC, splenocytes, lymph node cells…), both from Human or other species specimens, including non-Human primates and mice.

Main applications for immunomonitoring:

  • Enumeration and characterization of immune cell subsets (T, B, NK, DC, Mono.)
  • Characterization of T cell subset compartment (naïve, effector, central memory, effector memory)
  • Phenotypic and functional characterization of dendritic cell subsets (plasmacytoid and myeloid DC).
  • Biomarker expression analyses on various immune cell subsets
  • Counting of Ag specific T cells using tetramer technology
  • Cytokine production assessment by intracellular staining
  • Cytotoxicity analysis (CD107 mobilization assay, conjugate analysis…)
  • Cell proliferation analysis (BrDU uptake, CFSE)
  • Receptor saturation assays
     

Equipment: 2 FACS canto® II Analyzers (from Becton Dickinson). Both are calibrated every day, one serving as a backup for the other to prevent any trouble and spare critical labile samples.


Singulex Erenna

 

High sensibility:

When standard analytical technologies don’t reach appropriate signal/threshold resolution, high sensibility and/or very specific matrix analysis are required.

Main application:

  • detect low concentration (and potential variation) of soluble analytes in patient sera, or
  • quantification of cytokines baselines.

 

Multiplex Bead Array Assays (Luminex)

A flow cytometry-based technique allowing simultaneous measurement of a large number (up to several dozens) of analytes (cytokines, chemokines and many other markers) in a small volume (< 50 µL) of biological fluids or culture supernatants.

Main applications for immunomonitoring:

  • Large panels for cytokines and chemokines’ quantification (immune response, inflammation, cytokine storm)
  • Large panels for biomarkers’ analysis

Equipment: Bio-Plex® 200 (from Luminex).


ELISA

The Enzyme Linked ImmunoSorbent Assay is the gold-standard for the assessment of the concentration of soluble proteins in biological fluids (serum, plasma, urine) or cell culture supernatants.

Main applications for immunomonitoring:

  • Cytokine/chemokine dosage
  • Biomarker dosage
  • Ligand binding assays
  • Anti-drug antibody detections (ADA)
  • Auto-antibody detection


Equipment: Infinite® 200F (from Tecan).

 

Electrochemoluminescence Assays (i.e. MesoScale Discovery®)

MSD technology takes advantage of the electro-chemo-luminescence to offer ELISA-like assays with improved sensitivity and extended dynamic range. Thanks to its patented MULTI-ARRAY® technology, up to ten analytes of interest can be simultaneously quantified, sparing material from rare and critical samples.
 

Main applications for immunomonitoring:

  • Immunogenicity testing (dosage of ADA)
  • Multiple cytokine/chemokine concentration analysis (immune response, inflammation, cytokine storm)
  • High-throughput screenings


Equipment: Sector Imager 2400 plate reader.
 

 

ELISpot

An ELISA-derived method to detect and count protein secreting cells that is used mainly to enumerate antigen-specific lymphocytes or cytotoxic cells. Various cytokines (IFNg, IL-4, IL-10, IL-13, etc.) or other proteins (antibodies, granzyme, perforine, etc.) secretion can be assessed. Cells of interest are stimulated in vitro in the presence of various stimuli (specific antigenic protein or peptide, mAbs, target cells). Responsive cells are detected as spots which are enumerated using ELISpot plate reader and appropriate software.

Main applications for immunomonitoring:

  • Enumeration of antigen-specific cytokine-secreting T cells (vaccine development, autoimmunity, infectious disease, cancer, transplantation)
  • Enumeration of antigen-specific or cell specific cytotoxic cells (CTL, NK cells)
  • Enumeration of antibody producing B cells


Equipment: ImmunoSpot® Series 5 Micro Analyzer (from CTL)


Biobanking

ABL offers also biological sample storage services to its customers.

Properly registered in ABL’s sample database, the samples can be stored at different temperatures :-196°C in Liquid Nitrogen, in -80°C or -20°C freezers, or +4°C fridges.
All storage devices are securely locked and 24/7 temperature controlled, with emergency and backup organization.

ABL through its partnerships with logistics specialists has developed a capacity to centralize sample management, ensuring a full traceability from the collection in clinical investigation centers to the delivery to its laboratory or other sites.