Effect of deoxyguanosine on lymphopoiesis in the developing thymus rudiment <i>in vitro</i>: Application in the production of chimeric thymus rudiments

Jenkinson, Eric J.; Franchi, L. L.; Kingston, Rosetta; Owen, J. J. T.

doi:10.1002/eji.1830120710

Cited by 309 publications

(184 citation statements)

References 20 publications

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“…21 FTOCs were prepared as described previously 32 and depopulated in the presence of 1.5 mM 2dGuo in six-well tissue culture dishes. After 5-day depopulation, FTOC filters were moved to media without 2dGuo 33 for 1 day and then each FTOC lobe was reconstituted with 1-3 Â 10 4 retrovirally infected (24 h post-infection) cKit þ HPCs in a well of a Terasaki plate. After 24 h, the reconstituted FTOCs were transferred to a fresh filter and left to culture at 371C for 7-17 days with one media change after 7-10 days.…”

Section: Discussionmentioning

confidence: 99%

Tnfaip8 is an essential gene for the regulation of glucocorticoid-mediated apoptosis of thymocytes

et al. 2009

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Glucocorticoids have significant immunoregulatory actions on thymocytes and T cells and act by binding and activating cytosolic glucocorticoid receptors, which translocate to the nucleus and control gene expression through binding to specific response elements in target genes. Glucocorticoids promote cell death by activating an apoptotic program that requires transcriptional regulation. We set out to identify genes that are crucial to the process of glucocorticoid-mediated thymocyte apoptosis. Freshly isolated murine primary thymocytes were treated with dexamethasone, mRNA isolated and used to screen DNA microarrays. A set of candidate genes with upregulated expression was identified and selected members assayed in reconstituted fetal thymic organ culture (FTOC). Fetal liver-derived hematopoietic progenitor cells (HPCs) were infected with retroviruses expressing individual genes then used to repopulate depleted fetal thymic lobes. Reconstituted FTOCs expressing the gene Tnfaip8 were treated with dexamethasone and shown to be greatly sensitized to dexamethasone. Retrovirus-mediated RNA interference was applied to knock down Tnfaip8 expression in HPCs and these were used to reconstitute FTOCs. We observed that downregulating the expression of Tnfaip8 alone was sufficient to effectively protect thymocytes against glucocorticoid-induced apoptosis. We propose that Tnfaip8 is crucial in regulating glucocorticoid-mediated apoptosis of thymocytes.

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Section: Discussionmentioning

confidence: 99%

Tnfaip8 is an essential gene for the regulation of glucocorticoid-mediated apoptosis of thymocytes

et al. 2009

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“…We cultured the thymic lobes in the presence of dGuo, according to a previously described method with some modifications [43]. In brief, we placed Millipore filters (0.45-?…”

Section: Dguo Treatment Of Thymus Organ Culturementioning

confidence: 99%

Heterozygous null mutation of myelin P0 protein enhances susceptibility to autoimmune neuritis targeting P0 peptide

et al. 2003

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Mice with a heterozygous null mutation in myelin protein zero (P0 +/-) develop late-onset clinical paralysis associated with inflammatory pathology in the peripheral nerves. Although the development of this illness is known to require T cells and macrophages, little is understood regarding the immunological defect in the mice. Here we report that young P0 +/-mice, free from clinical manifestations, have a defect in central tolerance to P0, and are more prone to induction of experimental autoimmune neuritis (EAN) by sensitization against P0 180-199 peptide. Notably, we found that the P0 gene is transcribed in the thymus of wild-type and the P0 +/-mice in an amount proportional to the gene dosage. We then replaced the thymus of wild-type mice with that of the P0-deficient mice and vice versa. Immunization of these mice with P0 [180][181][182][183][184][185][186][187][188][189][190][191][192][193][194][195][196][197][198][199] revealed that a lower thymic P0 transcript would be associated with the higher recall T cell response to P0 [180][181][182][183][184][185][186][187][188][189][190][191][192][193][194][195][196][197][198][199] , thus accounting for the higher susceptibility of the P0 +/-mice to P0-induced EAN. These results imply that a heterozygous mutation in an autoantigen could cause defective central tolerance to the autoantigen. As such, autoimmune T cells may play some role in "genetic" diseases caused by a heterozygous gene defect.

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“…We constructed mouse chimeras to examine whether C10.4 + T cells could be positively selected on TEC. B6.PL (Thy-1.1 + ) day-16 fetal thymi were cultured in deoxyguanosine-containing medium to delete BM-derived cells [35], and transplanted under the kidney capsules of C10.4 + TAP -/--RAG -/-(C10.4 + T -R -) Thy1.2 + mice. After 3 months, mature-type C10.4 + CD8a + SP Thy-1.2 + T cells were detected in grafted thymi (Fig.…”

Section: Introductionmentioning

confidence: 99%

Maturational stage-dependent thymocyte responses to TCR engagement

Kattman¹,

Lukin²,

Oh³

et al. 2005

Eur. J. Immunol.

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Thymocyte positive and negative selection are dependent on avidity-driven TCRmediated recognition events in the thymus. High-avidity recognition events result in negative selection, while low-avidity recognition events result in positive selection. However, it has not been established how thymocytes maturation stages affect their responses to TCR signals of different avidities. We gained insight into this question when we reduced thymocyte selection to an in vitro system, in which full maturation of developmentally synchronized immature double-positive thymocytes was induced on a cloned line of thymic epithelial cells. Our analysis of the kinetics of thymocyte development supports a multi-phasic model of thymic selection. In it, thymocyte maturation stages as well as interaction avidity control the outcome TCR stimulation. Positive selection is initiated during a primary recognition event that proceeds independently of the TCR avidity. During a secondary recognition event the final fate of thymocyte, full maturation versus negative selection, is determined by TCR avidity.

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Effect of deoxyguanosine on lymphopoiesis in the developing thymus rudiment in vitro: Application in the production of chimeric thymus rudiments

Cited by 309 publications

References 20 publications

Tnfaip8 is an essential gene for the regulation of glucocorticoid-mediated apoptosis of thymocytes

Tnfaip8 is an essential gene for the regulation of glucocorticoid-mediated apoptosis of thymocytes

Heterozygous null mutation of myelin P0 protein enhances susceptibility to autoimmune neuritis targeting P0 peptide

Maturational stage-dependent thymocyte responses to TCR engagement

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