<i>In vitro</i> negative selection of αβ T cell receptor transgenic thymocytes by conditionally immortalized thymic cortical epithelial cell lines and dendritic cells

Tanaka, Yujiro; Mamalaki, Clio; Stockinger, Brigitta; Kioussis, Dimitris

doi:10.1002/eji.1830231035

Cited by 33 publications

(21 citation statements)

References 39 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The expression profile of both ␤-galactosidase and proinsulin (both Ins1 and Ins2) in the thymus and pancreas of these mice has been previously described (8,13). These were crossed with mice homozygous for the Immortomouse transgene, a temperature-sensitive SV40 large T-antigen, which induces proliferation of thymic epithelial cells at 33°C (22)(23)(24). At 37°C, these cells revert to a more differentiated phenotype (22)(23)(24).…”

Section: Methodsmentioning

confidence: 99%

“…These were crossed with mice homozygous for the Immortomouse transgene, a temperature-sensitive SV40 large T-antigen, which induces proliferation of thymic epithelial cells at 33°C (22)(23)(24). At 37°C, these cells revert to a more differentiated phenotype (22)(23)(24). Mice carrying this transgene against the C57Bl6 background were purchased from Charles River (Saint Constant, PQ, Canada) and have been previously described by Jat et al (22).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Isolation and Characterization of Proinsulin-Producing Medullary Thymic Epithelial Cell Clones

et al. 2006

View full text Add to dashboard Cite

Proinsulin, like many tissue-specific antigens, is expressed by rare (1-3%) cells of the thymus medullary stroma, presumably for the purpose of self-tolerance. Levels of this expression are associated with type 1 diabetes susceptibility in humans and in the NOD mouse. To further understand the mechanism of central tolerance induction by these rare cells, we have isolated and cultured two proinsulin-producing epithelial cell clones from murine thymus. These cells have a typical epithelial morphology and, by flow cytometry, a surface phenotype representative of mature thymic medullary epithelial cells (G8.8. By RT-PCR, they express predominantly Ins2 as opposed to Ins1, as does whole thymus. Expression of the transcription factor Aire, implicated in enhancing promiscuous thymic expression of tissue-specific antigens, fell to very low levels after a few passages but increased 20-fold upon exposure to an agonistic antilymphotoxin B antibody, concurrent with 2.5-fold enhanced insulin expression. RNA of Pdx-1, Glut-2, and Gck was detectable by RT-PCR in whole thymus but not in the clones, suggesting thymic proinsulin expression is Pdx-1 independent and that Pdx-1, Glut-2, and Gck are likely expressed in the thymus as antigens, not as regulatory molecules. Diabetes 55:2595-2601, 2006

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Proinsulin-Producing Medullary Thymic Epithelial Cell Clones

et al. 2006

View full text Add to dashboard Cite

show abstract

“…Influenza nucleoprotein peptide (366)(367)(368)(369)(370)(371)(372)(373)(374) Townsend et al, 1985) that contains sequences coding for 0c1, 2, and 328-498 of the nucleoprotein from the influenza virus (the deletion removes amino acids 3-327). Third, the resulting plasmid was cleaved at the EcoRI site of the Bluescript polylinker and the SmaI site described in the first cloning step and an EcoRI-NruI fragment from the H2-K b promoter was inserted (Weiss et al, 1983 Antigen-presenting cells were prepared by a modified protocol published previously (Tanaka, 1993 …”

Section: Introductionmentioning

confidence: 99%

Tolerance in TCR/Cognate Antigen Double‐TransgenicMice Mediated by Incomplete Thymic Deletion andPeripheral Receptor Downregulation

Mamalaki

Murdjeva

Tolaini

et al. 1995

Journal of Immunology Research

Self Cite

View full text Add to dashboard Cite

“…Furthermore, this peptide was shown to inhibit apoptosis of transgenic thymocytes (19) in an in vitro suspension culture assay (22). Because the suspension culture could not support the development of immature thymocytes, we could not determine whether this inhibition would result in the rescue and complete maturation of thymocytes.…”

mentioning

confidence: 99%

Interactions with multiple peptide ligands determine the fate of developing thymocytes

Williams¹,

Tarazona²,

Wack³

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Thymocytes are positively or negatively selected depending on interactions between their T cell receptors (TCR) and peptides presented by major histocompatibility complex molecules. We have previously shown that apoptosis of thymocytes from an ␣␤ TCR-transgenic mouse (F5), induced by antigenic peptide, can be inhibited specifically by an antagonist peptide variant in an in vitro culture model. We have now extended these experiments by demonstrating that the antagonist peptide can inhibit natural negative selection of maturing thymocytes, induced by endogenously expressed antigen, in fetal thymic organ cultures (FTOC). This inhibition resulted in the rescue and maturation of thymocytes that would otherwise have been deleted. Mature T cells generated in these cultures were able to respond to antigen by producing limited quantities of interferon-␥, but unlike T cells from control FTOC, they required exogenous interleukin-2 to generate cytolytic effector cells. Interestingly, the antagonist peptide also accelerated the development of F5 thymocytes in the absence of the negatively selecting ligand. These data suggest that the developmental fate of a thymocyte may be determined by the recognition of multiple distinct peptide ligands during thymic selection. Alterations in the profiles of selecting peptides presented in the thymus would thus have profound effects on the size and autoreactive potential of the T cell repertoire generated.T lymphocytes are positively or negatively selected in the thymus as a result of interactions between their T cell receptors (TCR) and thymic peptide-major histocompatibility complexes (MHC) (1-11). Much of the recent work on thymic selection was influenced by experiments that examined T cell responses to peptide analogs, produced from the antigenic peptide by substitution of amino acid residues involved in interaction with the TCR. Such peptide analogs can generate qualitatively different T cell responses compared with those produced by the antigenic peptide (12). In particular, some analogs were shown to act as TCR antagonists and inhibit T cell responses to the antigenic peptide (13). Several studies have shown that antagonist peptides are capable of positively selecting (9, 10), negatively selecting (14, 15), or blocking positive selection (16) of thymocytes. However, other studies have found that the ability of a peptide to select thymocytes positively or negatively does not always correlate with its properties as a TCR antagonist (10,11,17,18).We have previously reported an analog peptide (19) that was capable of inhibiting antigen-specific lysis of target cells by cytotoxic T lymphocyte (CTL) from ␣␤ TCR (F5)-transgenic mice (20,21). Furthermore, this peptide was shown to inhibit apoptosis of transgenic thymocytes (19) in an in vitro suspension culture assay (22). Because the suspension culture could not support the development of immature thymocytes, we could not determine whether this inhibition would result in the rescue and complete maturation of thymocytes. In the pre...

show abstract

In vitro negative selection of αβ T cell receptor transgenic thymocytes by conditionally immortalized thymic cortical epithelial cell lines and dendritic cells

Cited by 33 publications

References 39 publications

Isolation and Characterization of Proinsulin-Producing Medullary Thymic Epithelial Cell Clones

Isolation and Characterization of Proinsulin-Producing Medullary Thymic Epithelial Cell Clones

Tolerance in TCR/Cognate Antigen Double‐TransgenicMice Mediated by Incomplete Thymic Deletion andPeripheral Receptor Downregulation

Interactions with multiple peptide ligands determine the fate of developing thymocytes

Contact Info

Product

Resources

About