Institut National De La Sante Et De La Recherche Medicale

Txcell

Immunosearch

Txcell S.a.

Françoise Cottrez

Hervé Groux

Arnaud Foussat

Valérie Brun

Abdelilah Wakkach

Hervé Bastian

Nathalie Alepee

Virginie Leblanc

The present invention concerns an in vitro test for detecting the irritant potential of chemicals combining a corneal cell model with a selection of predictive and qualitative molecular markers to classify compounds into 3 categories, namely irreversible eye damage 21 days after application (category 1), reversible eye damage 21 days after application (category 2) and no irritation (no category). The inventors have thus demonstrated that the response following the action of an irritant substance occurs directly on an in vitro reconstructed corneal epithelium, and that the degree of irritation and the qualification of this irritation of a molecule may be determined by the use of specific biomarkers of eye irritation.

Method for evaluating the eye irritation potential of chemicals

The invention relates to a method for evaluating the sensitizing potential of a test compound, and to a kit for implementing said method.

Method for evaluating the sensitizing potential of a test compound

A method for the in vitro production of a cell population P' from a cell population P, the production requiring the presence of at least one factor which is expressed by feeder cells, wherein a) feeder cells proliferate at a temperature T, b) proliferated feeder cells are contacted with the cell population P, c) the cell mixture obtained at step (b) is cultivated at a temperature Twhich is chosen such that the cell population P proliferates and the feeder cells do not proliferate, the at least one factor being expressed by the feeder cells, and d) the cell population P&#x2032; so produced is recovered. Advantageously, the production consists in an expansion, the feeder cells are insect feeder cells and the cell population P to be expanded is a T lymphocyte population, preferably a Trl lymphocyte population.

In vitro production of a cell population using feeder cells

The invention relates to a method for preparing antigen-specific Tr1 regulatory lymphocytes. The inventive method involves the use of artificial antigen-presenting cells expressing a molecule from the HLA class II system and a human LFA-3 molecule and expressing none of the B7-1, B7-2, B7-H1, CD40, CD23, or ICAM-1 costimulatory molecules.

Method for obtaining antigen-specific Tr1 regulatory lymphocytes

An in vitro method for the obtention of a food- or auto-antigen specific Tr1 cell population from a leukocyte or a PBMC population, includes stimulating the PBMC or leukocyte population with the food- or auto-antigen, and recovering the food- or auto-antigen specific Tr1 cell population from the stimulated cell population. Preferably, the PBMC or leukocyte population is re-stimulated at least once with the same antigen after step (), in the presence of IL-2 and at least one interleukin selected from the group consisting of IL-4 and IL-13. The in vitro method may further include a third step of expanding the recovered antigen-specific Tr1 cell population, advantageously by contacting them with feeder cells capable of expressing factors necessary for the expansion. Preferably, the feeder cells are recombinant insect feeder cells.

Growing community of inventors

Le Rouret, France

Françoise Cottrez