Vα gene replacement in a TCRα knock-in mouse

Golub, Rachel; Huang, Ching-Yu; Kanagawa, Osami; Wu, Gillian E.

doi:10.1002/1521-4141(2001010)31:10<2919::aid-immu2919>3.0.co;2-n

Cited by 8 publications

(3 citation statements)

References 47 publications

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“…Endogenous TCR expression in the 5CC7 RAG-2-deficient mice could be mediated either by RAG-dependent recombination [13], gene conversion [14] or gene replacement [15] mechanisms involved in the generation of BCR and TCR diversity. To discriminate between these possibilities, we sequenced some of the TCRα and TCRβ-chains used by T-cells recovered from the allogeneic hosts.…”

Section: Resultsmentioning

confidence: 99%

Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection

et al. 2010

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Recombination activating gene (RAG)-deficient TCR (T Cell Receptor) Tg (transgenic) mice are routinely used as sources of monoclonal T cells. We found that after the transfer of T cells from a RAG-2-deficient 5CC7 TCR Tg mice into allogeneic hosts we recovered a population of T cells expressing diverse αβ-TCRs. In fact, in the thymus and spleen of the 5CC7 RAG-2-deficient donor mice, we detected rare T cells expressing non-Tg TCR chains. Similar observations were obtained using T cells from two other TCR transgenic strains, namely RAG-2-deficient aHY and RAG-1-deficient OT-1 mice. The sequences of the endogenous TCR transcripts suggested that gene recombination could occur, albeit quite inefficiently, in the RAG-deficient mice we used. In agreement, we evidenced rare TCR Vα and Vβ-chain transcripts in non-Tg RAG-2-deficient mice. Since in these non-Tg RAG-deficient mice no mature T cells could ever be found, our findings suggested a role for the TCR Tg in rescuing rare recombined endogenous chains. Robust T-cell activation by the allogeneic environment favored the selection and expansion of the rare cells expressing endogenous TCRs. Potential mechanisms involved in the recombination of the endogenous TCR chains in the different strains of RAG-deficient mice used, and in particular the possibility of RAG-1 hypomorphism due to an incomplete knocking out procedure, are discussed. Our findings have important experimental implications for studies using TCR-Tg RAG-deficient cells as monoclonal T cell populations.

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Section: Resultsmentioning

confidence: 99%

Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection

et al. 2010

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“…This mechanism had already been described in receptor revision of both immunoglobulins and T cells (Golub, 2002;Golub et al, 2001;Itoh et al, 2000), although in this latter case it was limited to the TCR␣ locus. To our knowledge, this is the first report in which evidence of V-gene replacement, although indirect, is presented in the case of TCR␤.…”

Section: Tcr Revision In At Heterozygotesmentioning

confidence: 61%

Analysis of Secondary V(D)J Rearrangements in Mature, Peripheral T Cells of Ataxia-Telangiectasia Heterozygotes

Lantelme

Turinetto

Mantovani

et al. 2003

Laboratory Investigation

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SUMMARY:Ataxia-telangiectasia (AT) is a rare recessive disease with pleiotropic involvement of the nervous and lymphoid systems. AT heterozygotes have a population frequency of about 1%, and although not manifesting any overt clinical symptoms, they have an increased mortality, mainly because of cancer and ischemic heart disease. We and others have described a mature T lymphocyte population with an altered T cell receptor surface expression ("TCR variant") that reactivates the recombination activating genes (RAG) and is expanded in the blood of patients with AT. In view of the known role of V(D)J recombination in the onset of tumorigenic translocations, we proposed that the increased RAG activity was responsible for the predisposition of AT homozygotes to develop mature-type T leukemia/lymphoma. In the present report, we used cytofluorimetry to quantify the TCR variant population and the memory/naïve T-cell compartments in the blood of AT heterozygotes compared with AT patients and controls. We assessed the expression of different recombinase genes through RT-PCR/oligotyping and cytofluorometric analysis and searched for rearrangement intermediates by ligase-mediated PCR in T-cell lines from four heterozygous carriers. We found the TCR variant population was increased on average 2ϫ in AT heterozygotes (vs 10ϫ in homozygotes) compared with controls, and naïve CD4 ϩ T lymphocytes were reduced on average 0.5ϫ (vs 0.1ϫ in homozygotes). We were able to demonstrate recombinase gene expression in all four heterozygous T-cell lines, and rearrangement intermediates, indicative of ongoing V(D)J recombination, in two. These rearrangements were compatible with V-gene replacement, a mechanism of receptor editing described for Ig and TCR␣ genes, to our knowledge not previously documented for TCR␤. In conclusion, we found that RAG reactivation and secondary V(D)J rearrangements, potential risk factors of mature-type leukemia in AT homozygotes, also take place in AT heterozygous carriers and might place this large population fraction at an increased risk of leukemia/lymphoma. (Lab Invest 2003, 83:1467-1475. E vidence supports the existence among peripheral T lymphocytes of a variant subpopulation with an altered TCR surface expression (CD4 ϩ CD3 low ) in which recombination activating genes (RAG) are expressed. We and others observed functional RAG gene products in mature T cells with defective TCR expression (Lantelme et al, 2000b;McMahan and Fink, 1998), and we showed that these T cells were increased in number in patients with ataxiatelangiectasia (AT) and Nijmegen breakage syndrome (Lantelme et al, 2000a), both autosomal recessive chromosomal instability syndromes characterized by immunodeficiency, hypersensitivity to ionizing radiation, defective DNA repair, and predisposition to develop lymphoid malignancies (Lavin and Shiloh, 1997;Shiloh, 1997). The finding of RAG gene expression in this TCR variant subpopulation suggests that secondary V(D)J rearrangements may occur physiologically to rescue their defective phenotype a...

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“…Inefficient positive selection has been shown to result in the replacement of transgenic TCRα chains with α chains from the endogenous locus [25], [26]. It has also been shown that a rearranged VαJα gene segment that was introduced into the genomic Jα region was removed by secondary TCRα rearrangements when the selecting MHC allele was not present [27], [28]. We were not able to directly measure the loss of cell surface expression of the transgene encoded TCRα chain since a mAb specific for this Vα gene segment does not exist and a published I-A u :Ac1-11 tetramer was no longer available.…”

Section: Resultsmentioning

confidence: 99%

TCR Affinity for Self-Ligands Influences the Development and Function of Encephalitogenic T Cells

Vandal

Sant’Angelo

2011

PLoS ONE

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The specificity and affinity of self-reactive T cells is likely to impact the development of autoimmune-disease causing T cells in the thymus as well as their function in the periphery. We identified a naturally occurring, low affinity variant of an MBP Ac1-11/I-Au specific TCR that is known to induce EAE. Thymocytes in mice carrying the transgenes for this low affinity TCR were poorly positively selected, as compared to their high affinity TCR expressing counterparts. Nonetheless, CD4 T cells bearing the low affinity TCR accumulated in the periphery of the mice. Unlike mice expressing the high affinity TCR, these mice very rarely developed disease. However, if endogenous TCR expression was eliminated by breeding to RAG1 deficient mice, 100% of the mice carrying either the high or the low affinity versions of the TCR developed EAE. Intriguingly, while the incidence of EAE increased, the age of onset of disease in both mice was identical. These data suggest disease onset occurs during a short window of mouse development.

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Vα gene replacement in a TCRα knock-in mouse

Cited by 8 publications

References 47 publications

Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection

Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection

Analysis of Secondary V(D)J Rearrangements in Mature, Peripheral T Cells of Ataxia-Telangiectasia Heterozygotes

TCR Affinity for Self-Ligands Influences the Development and Function of Encephalitogenic T Cells

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