Our in vitro macrophage-based assays are relevant to assess immunotherapeutics and question their mechanisms of action.
In the context of cancer immune surveillance, dendritic cells (DCs) are key immune cells as they possess a unique ability to prime and expand antigen-specific CD4 and CD8 T cells. While trafficking from the tumor to the draining lymph nodes, they can cross-present tumor antigens and initiate a specific anti-tumor immune response. Even though several subsets exist, DCs must receive an activating signal to initiate a process of maturation that converts them from an accumulation to an antigen presentation mode to finally promote immunity or tolerance depending on the nature of the stimulus.
Thus, considering the importance of DCs in fine-tuning T cell response, their manipulation represents an attractive approach to elicit or boost an anti-tumor immune response. For this purpose, benefiting from a robust assay that evaluates the function and immunomodulation of DCs is highly valuable.
To this end, we have developed and validated a mixed leukocyte reaction (MLR) assay which is based on a co-culture of allogenic CD4+ T cells (responder) and monocyte-derived DCs (stimulator). The data shown here, originating from independent donor pairs, illustrate stimulatory or immunosuppressive properties of reference compounds. Indeed, while Adenosine, a well-known immunomodulatory nucleoside, favored a “tolerogenic” profile that translated into a limited MLR response as shown through lower levels of cytokines released in the co-culture, Nivolumab application significantly boosted the MLR response.
Hence, this dataset underlines the effectiveness of our assay in addressing the ability of test items to modulate DC-mediated immune responses.
MLR in allogenic human mDC/CD4 T cell co-cultures from independent donor pairs is enhanced upon PD1 inhibition and limited by Adenosine.
Isolated peripheral blood monocyte-derived DCs are co-cultured with allogenic CD4+ T cells. At the end of the co-culture period, IFNg (A) and IL2 (B) levels released in the supernatants are measured by HTRF.
While mature DCs display T-cell stimulating ability underlined through the MLR response, this response is further optimized following PD1 blockade with Nivolumab and limited upon Adenosine treatment.
More about our MLR assays
Thus, considering the importance of DCs in fine-tuning T cell response, their manipulation represents an attractive approach to elicit or boost an anti-tumor immune response. For this purpose, benefiting from a robust assay that evaluates the function and immunomodulation of DCs is highly valuable.
To this end, we have developed and validated a mixed leukocyte reaction (MLR) assay which is based on a co-culture of allogenic CD4+ T cells (responder) and monocyte-derived DCs (stimulator). The data shown here, originating from independent donor pairs, illustrate stimulatory or immunosuppressive properties of reference compounds. Indeed, while Adenosine, a well-known immunomodulatory nucleoside, favored a “tolerogenic” profile that translated into a limited MLR response as shown through lower levels of cytokines released in the co-culture, Nivolumab application significantly boosted the MLR response.
Hence, this dataset underlines the effectiveness of our assay in addressing the ability of test items to modulate DC-mediated immune responses.
MLR in allogenic human mDC/CD4 T cell co-cultures from independent donor pairs is enhanced upon PD1 inhibition and limited by Adenosine.
Isolated peripheral blood monocyte-derived DCs are co-cultured with allogenic CD4+ T cells. At the end of the co-culture period, IFNg (A) and IL2 (B) levels released in the supernatants are measured by HTRF.
While mature DCs display T-cell stimulating ability underlined through the MLR response, this response is further optimized following PD1 blockade with Nivolumab and limited upon Adenosine treatment.
More about our MLR assays
Discover how to fully exploit the mixed leukocyte reaction assay for assessing immunotherapeutics on dendritic cell function and mediated immune response.
Ovarian cancer ascites are a remarkable opportunity to improve your translational research and therapeutic testing!
As Olink®-certified service Provider, Explicyte offers now high proteomic analysis capabilities for comprehensive protein biomarker discovery using Olink® Target technology.
The GeoMx Whole Transcriptome Atlas (WTA) empowers to explore biology accross a tissue. The human whole transcriptome (over 18,000 protein-coding human genes) is measured in each region of interest to uncover biological changes at specific locations of the tissue.
The GeoMx Whole Transcriptome Atlas delivers the maximum amount of sensitivity and confidence in each transcript through its unique probe architecture. The WTA profiles over 18,000 protein-coding human genes based on the human gene nomenclature committee (HUGO1) database cross-referenced with available mRNA sequences in the National Center for Biotechnology’s Information (NCBI) RefSeq database.
The WTA then allows to explore pathways across the whole transcriptome in defined regions of interest.
- Whole transcriptome coverage with probes specific to protein coding mRNA sequences
- Compatible with common sample types such as formalin-fixed paraffin embedded (FFPE) or fresh frozen (FF) tissue and across all human tissues types
- Superior sensitivity to detect 1000s of unique human genes in <50 μm regions
- Robust performance across sample types including FFPE with high concordance with RNA-seq and RNAscope™
- Map single cell RNA-seq populations to their tissue location
The GeoMx Whole Transcriptome Atlas delivers the maximum amount of sensitivity and confidence in each transcript through its unique probe architecture. The WTA profiles over 18,000 protein-coding human genes based on the human gene nomenclature committee (HUGO1) database cross-referenced with available mRNA sequences in the National Center for Biotechnology’s Information (NCBI) RefSeq database.
The WTA then allows to explore pathways across the whole transcriptome in defined regions of interest.
Learn about out Digitial Spatial Profiling (DSP) services
The Cancer Transcriptome Atlas (CTA) is designed for comprehensive profiling of the tumor biology, tumor microenvironment, and the immune response. It covers the RNA expression of over 1,800 genes simultaneously with spatial resolution in any region of interest from a single tissue section.
The CTA assay is compatible with RNAscope® and antibody visualization markers. It can also be supplemented with up to 60 additional targets of interest.
- Extensive coverage of the immune response, tissue microenvironment, tumor biology, and genes from clinically relevant genes sets such as tumor inflammation
- Spatial measurement of single cell signatures with high sensitivity and dynamic range
- Inclusion of genes for the Tumor Inflammation Signature, PAM50 and other clinical signatures
- Over 100 pathways to explore all aspects of cancer and tumor biology
Learn about out spatial transcriptomics services
Comprehensively Annotated Pathways in the CTA
| Adaptive Immunity | | Cell Function | | Signaling Pathways |
| T cells B cells | Apoptosis | AMPK Signaling | ||
| TCR & BCR Signaling | Autophagy | Androgen Signaling | ||
| Cancer Antigens | Cell Adhesion & Motility | EGFR Signaling | ||
| MHC Class I & II Antigen Presentation | Cell Cycle | ERBB2 Signaling | ||
| T-cell Checkpoints | Cilium Assembly | Estrogen Signaling | ||
| TH1, TH2, TH9, Th17, and Treg Differentiation | Differentiation | FGFR Signaling | ||
| DNA Damage Repair | FoxO Signaling | |||
| Innate Immunity | EMT | GPCR Signaling | ||
| Complement System | Endocytosis | Hedgehog Signaling | ||
| Dendritic Cells | Epigenetic Modification | HIF1 Signaling | ||
| DNA & RNA Sensing | Immortality & Stemness | Insulin Signaling | ||
| Glycan Sensing | Ion Transport | JAK-STAT Signaling | ||
| Host Defense Peptides | Lysosome | MAPK Signaling | ||
| Inflammasomes | Oxidative Stress | MET Signaling | ||
| Myeloid Inflammation | Phagocytosis | mTOR Signaling | ||
| Neutrophil Degranulation | Proteotoxic Stress | Myc | ||
| NK Activity | RNA Processing | NO Signaling | ||
| NLR Signaling | Senescence | Notch Signaling | ||
| RAGE Signaling | p53 Signaling | |||
| TLR Signaling | Metabolism | PDGF Signaling | ||
| Amino Acid Synthesis & Transport | PI3K-Akt Signaling | |||
| Immune Response | Arginine & Glutamine Metabolism | PPAR Signaling | ||
| Chemokine Signaling | Fatty Acid Oxidation & Synthesis | Purinergic Signaling | ||
| Cytotoxicity | Glycolysis & Glucose Transport | Retinoic Acid Signaling | ||
| IL-1, IL-2, IL-6 & IL-17 Signaling | Glycosylation | TGF-beta Signaling | ||
| Immune Exhaustion | IDH1/2 | VEGF Signaling | ||
| Interferon Response Genes | Lipid Metabolism | Wnt Signaling | ||
| Lymphocyte Regulation & Trafficking | Mitochondrial Metabolism / TCA | |||
| NF-kB Signaling | Nucleotide Synthesis | Physiology & Disease | ||
| Other Interleukin Signaling | Pentose Phosphate | Angiotensin System | ||
| Prostaglandin Inflammation | Tryptophan & Kynurenine Metabolism | Circadian Clock | ||
| TNF Signaling | Vitamin & Cofactor Metabolism | Drug Resistance | ||
| Type I, II, & III Interferon Signaling | | Glioma | ||
| | Leukemia | |||
| Matrix Remodeling and Metastasis | ||||
| Melanoma | ||||
| Neuroendocrine Function | ||||
| Prostate Cancer |
Soft Tissue Sarcoma (STS) is known to be refractory to current cancer immunotherapies including the PD1 immune checkpoint inhibitor (ICI) (Toulmonde et al., 2018), thus claiming the development of novel therapeutic strategies aiming at improving the current clinical benefit.
Drawing on our expertise in IHC and IHF assays, we have developed and validated a series of markers & panels to assess several markers of interest as well as the abundance and spatial distribution of lymphoid and myeloid immune cell subsets.
Discover why there is a crucial need to decipher mechanisms underlying sensitivity / resistance to cancer immunotherapies which can ultimately lead to the identification of novel therapeutic targets and how to achieve it.
Our data illustrated here through DC differentiation and mixed leukocyte reaction assays show that adenosine-mediated suppression results in the generation of phenotypically and functionally dysregulated DCs, and that this process is relieved by adenosine receptor antagonists.
We have fully set up and characterized a syngeneic Lewis LLC1 mouse model for preclinical studies of novel therapeutic agents and anti-cancer strategies.