Immunoprofiling

Purpose

Controlling infectious diseases requires understanding of the relationships between immune cells and pathogens.

Most current studies to understand infectious processes rely on models requiring

• Infection of cell culture or explants
• Analysis of pathological phenotypes induced after prior fixation of infected samples obtained from infected mice. The possibility of carrying out those manipulations in a protected confined environment avoids the tissues fixation steps which often impair the possibility of obtaining high-quality anatomical and cellular information. However, very few laboratories are capable of carrying out these experiments.

The Biosafety Level 3 Containment Facility (BSL3) has special engineering and containment features that allow investigators to work safely with pathogens classified as risk group 2 or 3. This facility consists of three laboratory suites corresponding to an animal housing room, a laboratory for in vitro experiments and a bio-imaging area.

Equipments

• Through PHENOMIX program, we equipped the BSL3 module with a multiphoton confocal microscope that is specially designed to operate in such a confined environment.
• The Infrastructure program PHENOMIN enabled us to acquire a small animal imaging bioluminescence system IVIS III.
• Finally, the Infectiopole-Sud program provided us a mouse housing cage system adapted to BSL3 conditions and a cryostat.

Phenotyping tools

All instruments are accessible to trained users based on the scientific and regulatory rules established by scientific and biosecurity and safety manager of BSL3. Although the BSL3 platform was only fully operational in May 2015, it already permitted to set up

• Vaccine assays,
• Real-time imaging of immune responses in mice challenged by pathogens,
• An infection-immunophenotyping analysis pipeline,
• To test the ability of adjuvants to enhance vaccine efficacy in a mouse model of infection.

Diseases investigation and models

Real-time imaging studies of immune responses in mice challenged by pathogens such as Salmonella or Brucella infection have led to the development of an immunophenotyping analytical pipeline devoted to infection in synergy with our Immunophenotyping module. The analysis of organ distribution of biomolecules engineered to transport and deliver nucleic acids on living animals using an IVIS Lumina III device is also developed in the frame of a contract with a biotech company. Therefore, we can visualize the spread of infection in vivo, isolate and characterize the pathogens, and analyze phenotypic and functional modification of immune system in the course of infections by advanced live-imaging and flow and mass cytometry.

Access and conditions of equipment use

We provide a broad range of instruments and services for academic and industrial customers. An experienced staff of scientists, technical experts and instrument specialist provides instructions for the design, execution and analysis of standardized multiparametric flow, spectral, mass cytometry and advanced microscopy based studies. All these cutting-edge equipment are technically and operationally enrolled in a regular maintenance program with their commercial providers. Moreover, instrument specialists at PHENOMIN are dedicated to the maintenance and tuning of the instruments on a daily basis. The characterization of the cytometer status and performance using CS&T beads for cytometer set up and tracking system, ensure reproducible performance and instrument sensitivity on a day to day basis. All experiments are standardized using applications setting in order to achieve: (1) high consistency of results over time and (2) facilitate data analysis and results validation. All data are archived and stored on dedicated servers. Depending on the technical complexity and availability, the equipment can be handled by our experts or made available for use to trained customers. PHENOMIN has set up an on line instrument tutorial for training external users. PHENOMIN applies two types of price: one for its academic clients on full cost basis and the other one for its industrial customers taking into consideration intellectual property and the transfer of know-how.

Purpose

Real-time imaging studies of immune responses in mice challenged by pathogens (Salmonella infection model) and infection-immunophenotyping analysis pipeline in synergy with immunophenotyping CIPHE facility.

Purpose

Set up vaccine assays and test the ability of adjuvants to enhance vaccine efficacy in a mouse model of infection.