Method for obtaining antigen-specific Tr1 regulatory lymphocytes
AssigneeInstitut National De La Sante Et De La Recherche Medicale (Inserm)
InventorsHervé Groux, Françoise Cottrez, Abdelilah Wakkach
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Abstract
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.
Claims8
A method of preparing antigen-specific regulatory Tr1 lymphocytes by activation and differentiation of CD4+ lymphocytes of a subject, comprising:
- separating lymphocytes from a sample of the subject's blood,
- activating said lymphocytes in vitro by co-culturing with artificial antigen-presenting cells comprising cells which express a class II molecule of the HLA system and a human LFA-3 molecule and which do not express any of the costimulatory molecules B7-1, B7-2, B7-H1, CD40, CD23 or ICAM-1, and which present a selected antigen associated to the class II molecule,
- recovering from said activated lymphocytes a population of activated CD4+ T lymphocytes comprising at least 10% of Tr1 lymphocytes which are specific for the antigen of the antigen-presenting cells, and
- isolating said Tr1 lymphocytes from said population of activated CD4+ T lymphocytes to produce said antigen-specific regulatory Tr1 lymphocytes.
A method of preparing antigen-specific regulatory Tr1 lymphocytes by activation and differentiation of CD4+ lymphocytes of a subject, comprising:
- separating lymphocytes from a sample of the subject's blood,
- activating said lymphocytes in vitro by co-culturing with artificial antigen-presenting cells comprising cells which express a class II molecule of the HLA system and a human LFA-3 molecule and which do not express any of the costimulatory molecules B7-1, B7-2, B7-H1, CD40, CD23 or ICAM-1, and which present a selected antigen associated to the class II molecule,
- recovering from said activated lymphocytes a population of activated CD4+ T lymphocytes comprising at least 10% of Tr1 lymphocytes which are specific for the antigen of the antigen-presenting cells,
- repeating the activating step by activating the recovered population of CD4+ T lymphocytes in vitro with said antigen-presenting cells, and enriching the population of activated CD4+ lymphocytes to at least 30% Tr1 lymphocytes that are specific for the selected antigen, and
- isolating said Tr1 lymphocytes from said population of activated CD4+ T lymphocytes to produce said antigen-specific regulatory Tr1 lymphocytes.
A method of preparing antigen-specific regulatory Tr1 lymphocytes by activation and differentiation of CD4+ lymphocytes of a subject, comprising:
- separating lymphocytes from a sample of a subject's blood,
- activating said lymphocytes in vitro by co-culturing with artificial antigen-presenting cells comprising cells which express a class II molecule of the HLA system and a human LFA-3 molecule and which do not express any of the costimulatory molecules B7-1, B7-2, B7-H1, CD40, CD23 or ICAM-1, and which present a selected antigen associated to the class II molecule,
- recovering from said activated lymphocytes a population of activated CD4+ T lymphocytes comprising at least 10% of Tr1 lymphocytes which are specific for the antigen of the antigen-presenting cells,
- isolating Tr1 lymphocyte clones from said recovered population of activated CD4+ T lymphocytes, and
- propagating the isolated Tr1 lymphocyte clones.
Dependencies are inferred from the claim text.