Retroviral transduction of lineage antigen-negative (Lin?) cells: A valuable alternative for the generation of T cell receptor (TCR) retrogenic mice
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- 作者：Christophe Viret ; Charlotte Cresson ; Cyril Broccardo ; Sylvie Guerder
- 关键词：Retrogenesis ; Hemopoietic stem cells ; T cell receptor
- 刊名：Journal of Immunological Methods
- 出版年：2012
- 出版时间：31 October, 2012
- 年：2012
- 卷：384
- 期：1-2
- 页码：103-110
- 全文大小：814 K
文摘
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 ¦Á and ¦Â 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 (Lin?) cells from untreated donors for TCR retrogenesis. In contrast to cells from 5-FU-treated donors, transduced Lin? 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. Lin? cell-enriched BM cells represent therefore, a reliable alternative to 5-FU treatment for retroviral transduction of haematopoietic stem cells and TCR retrogenic derivation.