Inflammation resolution is the necessary biological process to control inflammation.
This process is characterized by the sequential production of specialized pro-resolving mediators (SPMs) that allow to modulate the immune response, fight against infections and allow the return to homeostasis in tissues. The main families of SPMs are lipoxins, resolvins, protectins, maresins.
Ambiotis have at disposal various in vitro and in vivo experimental approaches to evaluate the efficacy of a product to activate the resolution of inflammation:
- Quantification of SPM by LC-MS/MS
- Evaluation of enzymes (COX, LOX) expression by RT-PCR
- Evaluation of phagocytosis and efferocytosis by real-time cell imaging
- Evaluation of MI/M2 macrophages differentiation by RT-PC or immuno-assay
In vitro
Models
IMMUNE CELLS
- Whole blood
- PBMC (monocytes/lymphocytes)
- Monocytes
- M1/M2 macrophages
- Foamy macrophages
- Atheromatous plaque macrophages
- Dendritic/Langerhans cells
- Lymphocytes (Th1, Th2, Th17…)
- Granulocytes
SKIN CELLS
- Keratinocytes
- Fibroblasts
- Preadipocytes- adipocytes
- Melanocytes
INTESTINAL CELLS
HEPATOCYTES
Read-outs
- Lipid mediators quantification
- Differentiation capacity (DC maturation, macrophage polarization…)
- Infectious agents elimination (Phagocytosis assays)
- Cells elimination (Efferocytosis assays)
Illustrative data
Phagocytosis model
Phagocytosis reflects the innate immune function of macrophages. The phagocytosis of E. coli bacteria by primary macrophages can be dynamically visualized in real-time using the Incucyte imaging system. This assay highlights the cellular capacity for pathogen clearance. These insights are essential for understanding immune responses in both physiological and pathological contexts.
Global purpose: Screen new molecules for pro-resolutive properties, most particularly on pathogens cleaning activity.
Characteristics: Live cell imaging and fluorescence monitoring during 6h
MI/M2 macrophages polarization model
Macrophage polarization represents a continuum between the two main M1 (inflammatory) and M2 (resolving) subtypes. At any given time point, the macrophage population shows a combination of macrophage phenotypes in a state of transition that express markers and functions of both MI and M2 phenotypes. In some pathophysiological contexts one subtype will be more preponderant than the other one. Thus, a balance in macrophage phenotypes plays a key role in homeostasis and pathogenesis.
Global purpose: Screen new molecules for pro-resolutive properties, most particularly on macrophage phenotype.
Characteristics: Quantification of gene expression characterizing M1 and M2 phenotypes
More data ? Contact our team
In vivo
Experimentals models
- Peritonitis model in mouse induced by zymosan or thioglycollate
Read-outs
- Kinetics of immune cell recruitment ( identification of leukocyte subpopulations by cytometry : monocytes, macrophages, neutrophils, lymphocytes, eosinophils, …)
- Resolution indices (magnitude, duration, resolution interval)
- Resolution signature (bioactive lipids quantification by LC-MS/MS)
- Inflammation signature (cytokine and other markers quantification by ELISA, Luminex or RT-PCR)
- Efferocytosis
Illustrative data
Zymosan-induced Peritonitis model
Pre-clinical models of peritonitis allow to assess pro-resolving activity of drugs. The inflammation is induced by injection of zymosan, into the peritoneum. Pro-resolving parameters can be monitored over time.
This model allows to quantify the recruitment of inflammatory cells and the production of cytokines and bioactive lipids. The inflammatory status (acute phase or resolution) can also be identified. Finally, macrophages can be collected from the peritoneal cavity to be further analyzed in cell culture for functional properties.
Global purpose: Evaluation anti-inflammatory and pro-resolutive properties of a compound by studying intensity and kinetics (PK/PD) of the inflammatory and resolution phases.
Caracteristics: Length : 24h
Species: Mice, rat
Ways of working with us
Your need
- Understanding of your need
- Orientation toward the relevant work package
Our collaboration
- Performance of services according milestones and go/no go steps
- Regular follow-up
Your results
- Scientific feedback through webex conference
- Final report
- Follow up