Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8418535 | Journal of Immunological Methods | 2012 | 8 Pages |
Abstract
Mice with virtually all T cells expressing a single T cell receptor (TCR) on their surface have been instrumental in understanding the development of immature thymocytes. For many years, such an engineering has been achieved essentially by inserting rearranged TCR α and β chain coding sequences into the genome through co-microinjection into fertilized eggs (TCR transgenesis). More recently, a novel methodology relying on the reconstitution of T cell deficient hosts with retrovirally-transduced multipotent bone marrow cells has been developed. Hence, TCR retrogenesis allows for the in vivo study of given TCR specificities in a faster and less expensive manner. While initial procedures were taking advantage of 5-Fluorouracil (5-FU) treatment of RAG-deficient or SCID donor mice as source of haematopoietic stem cells, we used bone marrow cell suspensions enriched in lineage antigen-negative (Linâ) cells from untreated donors for TCR retrogenesis. In contrast to cells from 5-FU-treated donors, transduced Linâ cells consistently generated a sizable retrogenic pool of thymocytes and required less donor mice. In such retrogenic mice, immature thymocytes bearing a major histocompatibility complex (MHC) class II-restricted TCR differentiated into the expected CD4 mature T cell lineage and populated the peripheral lymphoid organs where they retained the capacity to react to their cognate ligand. Linâ cell-enriched BM cells represent therefore, a reliable alternative to 5-FU treatment for retroviral transduction of haematopoietic stem cells and TCR retrogenic derivation.
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Authors
Christophe Viret, Charlotte Cresson, Cyril Broccardo, Sylvie Guerder,