Human immune-monitoring core
The human immune-monitoring core of the Physiopathology Center of Toulouse Purpan specializes in the immune characterization and the conservation of human immune cells. Our services focus on the analysis of the phenotype and the functional responses of immune cells from patient samples in the context of clinical studies in infection, autoimmunity or inflammation. In addition, we constitute and distribute a collection of PBMC, plasma and DNA from > 250 healthy subjects.
Lastly, we have an expertise in regulatory affairs related to human research and act as a administrative link in our institution.
Immune-monitoring on patient clinical samples
We carry out analyzes, including longitudinal, of samples from patients supervised at the University Hospital Centers of France. The samples come to us fresh or frozen. The analyses carried out are at the request of the investigators and include: standardized immune-phenotyping, in connection with the cytometry platform of the Center, standardized ELISPOT with the inclusion of control reference donors, or optimization of biobanking protocols.
Samples from patients with hepatitis E virus infection, multiple sclerosis, or narcolepsy were thus analyzed by the core facility.
Healthy subject PBMC biobanking
- PBMC are isolated from buffy coats provided by the “Établissement Français du Sang” (EFS) at the Purpan hospital in Toulouse, France
- The PBMC bank is from adult volunteers donating their blood for a therapeutic use and who are recruited in the greater Toulouse – Occitania region. Donors reported feeling well and were evaluated in good general health by a doctor at the EFS. Each blood donation was monitored for the absence of permanent contraindication for blood donation (viral hepatitis, syphilis, HIV or HTLV infection)
- PBMC are characterized for donor gender, age, blood type, HLA-A2 and HLA-B7 or -B8 expression, CMV and toxoplasmosis serology
- The core facility provides frozen PBMC vials, each containing 15 or 20 million PBMC. Plasma and DNA are also conserved for purposes of molecular analyses
- 50 to 110 new donors are added annually to the biobank in order to reach a constant availability of at least 250 donors
- At least 1 vial from each donor is thawed by the core facility and analyzed for PBMC recovery or viability. Other quality controls include the analysis of the production of interferon by memory CD8+ T cells.
- PBMC isolation procedure: PBMC isolation and cryopreservation
- PBMC thawing protocol: PBMC thawing
- PBMC request (only for CPTP members): r- PBMC request send form to contact
Development and standardization of immune assays on human cells
- This activity is carried out during 8-months rotations in laboratories involved in human immune-monitoring studies
- We provide support for the definition of project-specific immune assays, in addition we evaluate their inter-experimental variability across experimenters and over time
Example of projects : ex vivo ELISPOT assays for the measurement of memory T cell response specific for the hepatitis E virus, definition and testing of multiparametric flow cytometric profiles, isolation of rare Treg cell population circulating toward the thymus, handling of low-volume blood samples such as pediatric samples
Network with hospitals and human research regulatory affairs
We act as a liaison with several institutions’ departments of regulatory affairs to ensure that the center complies with the ethical and regulatory requirements for human research. Examples: writing of patient information sheets, informed consent forms, or administrative documents related to collections of human biological samples.
Examples include the setting up (ethics, administration and logistics) of transfers of human surgical wastes for research, the declaration of research patients with multiple sclerosis or narcolepsy.
Useful links to understand the rules of clinical research in France:
- Human research is controlled by legislative texts that were overhauled in 2016 with the application of the Jardé law (C. santé publ. Partie législative, Première partie : Protection générale de la santé, Livre Ier : Protection des personnes en matière de santé, Titre II : Recherches impliquant la personne humaine, Chapitre I à IV)
- Subject confidentiality and privacy must be ensured (Loi n° 78-17 du 6 janvier 1978 relative à l’informatique, aux fichiers et aux libertés). Most research must be authorized by or reported to the Commission Informatique et Liberté (CNIL): https://www.cnil.fr/fr/
- To seek endorsement of a clinical research project by the INSERM: https://www.inserm.fr/professionnels-recherche/recherche-sur-personnes/soumission-projets-impliquant-personne-humaine
- Guidelines from the French Ministère des Solidarités et de la santé https://solidarites-sante.gouv.fr/systeme-de-sante-et-medico-social/recherche-et-innovation/recherches-impliquant-la-personne-humaine/
- S6 Ultra-V analyzer (Cellular Technology Ltd)
- In the L2 lab – F building
- No fees
- Florence Abravanel
- Eric Champagne
- Manuel Diaz Munoz
- Loïc Dupré
- Nicolas Fazilleau
- Sylvie Guerder
- Jean-Charles Guéry
- Roland Liblau
- Xavier Iriart
- Lucette Pelletier
- Laurent Réber
- Emmanuel Treiner
- Joost van Meerwijk