RESUMO
Aging is associated with a gradual loss of naïve T cells and a reciprocal increase in the proportion of memory T cells. While reduced thymic output is important, age-dependent changes in factors supporting naïve T cells homeostasis may also be involved. Indeed, we noted a dramatic decrease in the ability of aged mice to support survival and homeostatic proliferation of naïve T cells. The defect was not due to a reduction in IL-7 expression, but from a combination of changes in the secondary lymphoid environment that impaired naïve T cell entry and access to key survival factors. We observed an age-related shift in the expression of homing chemokines and structural deterioration of the stromal network in T cell zones. Treatment with IL-7/mAb complexes can restore naïve T cell homeostatic proliferation in aged mice. Our data suggests that homeostatic mechanisms that support the naïve T cell pool deteriorate with age.
Assuntos
Homeostase/fisiologia , Memória Imunológica/imunologia , Ativação Linfocitária/imunologia , Tecido Linfoide/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Proliferação de Células , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Studies using glucocorticoid receptor exon 2-disrupted knockout (GR2KO) mice provided strong evidence against an obligatory role for glucocorticoid receptor (GR) signalling in T-cell selection. These mice express a truncated form of the GR that is incapable of transmitting a range of glucocorticoid (GC)-induced signals, including GC-induced thymocyte death. However, one study that suggested that truncated GR function is preserved in the context of GR-mediated repression of T-cell activation-induced genes, challenged earlier conclusions derived from the use of these mice. Because GR versus T-cell receptor (TCR) signalling cross-talk is the means by which GCs are hypothesized to have a role in T-cell selection, we reassessed the utility of GR2KO mice to study the role of the GR in this process. Here, we show that GR-mediated repression of TCR signalling is impaired in GR2KO T cells in terms of TCR-induced activation, proliferation and cytokine production. GC-induced apoptosis was largely abolished in peripheral T cells, and induction of the GC-responsive molecule, interleukin-7 receptor, was also severely reduced in GR2KO thymocytes. Together, these data strongly re-affirm conclusions derived from earlier studies of these mice that the GR is not obligatory for normal T-cell selection.
Assuntos
Seleção Clonal Mediada por Antígeno/fisiologia , Éxons , Glucocorticoides/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Receptores de Glucocorticoides/imunologia , Transdução de Sinais/imunologia , Animais , Glucocorticoides/genética , Ativação Linfocitária/fisiologia , Camundongos , Camundongos Knockout , Receptores de Antígenos de Linfócitos T/genética , Receptores de Glucocorticoides/genética , Transdução de Sinais/genética , Timo/imunologiaRESUMO
Cytomegalovirus (CMV) infection promotes a broad T-cell response, with the resulting memory cells displaying diverse phenotypes. CMV establishes lifelong persistence/latency, and it is thought that viral antigens expressed during this period may regulate the expansion and/or maintenance of "inflationary" CD8 T-memory populations that display an effector memory phenotype. We show here that mouse CMV (MCMV)-specific inflationary memory T cells do not decrease in number after thymectomy, indicating that recent thymic emigrants are not strictly required for their maintenance. Furthermore, persistent MCMV replication in the salivary gland does not significantly impact the T-cell memory compartment, as surgical removal did not alter its composition. These results shed light upon the mechanisms required for maintenance of the large, MCMV-specific T-cell memory pool.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Memória Imunológica , Muromegalovirus/imunologia , Animais , Camundongos , Timectomia , Fatores de TempoRESUMO
Although it is known that interleukin-7 (IL-7) and IL-15 influence the survival and turnover of CD8+ T cells, less is known about how these cytokines affect different subsets during the course of the immune response. We find that IL-7 and IL-15 differentially regulate CD8+ T-cell subsets defined by KLRG1 and CD127 expression during the contraction phase of the immune response. The provision of IL-15, or the related cytokine IL-2, during contraction led to the preferential accumulation of KLRG1(hi)CD127(lo) CD8+ T cells, whereas provision of IL-7 instead favored the accumulation of KLRG1(lo)CD127(hi) cells. While IL-7 and IL-15 both induced proliferation of KLRG1(lo) cells, KLRG1(hi) cells exhibited an extraordinarily high level of resistance to cytokine-driven proliferation in vivo despite their dramatic accumulation upon IL-15 administration. These results suggest that IL-15 and IL-2 greatly improve the survival of KLRG1(hi) CD8+ T cells, which are usually destined to perish during contraction, without inducing proliferation. As the availability of IL-15 and IL-2 is enhanced during periods of extended inflammation, our results suggest a mechanism in which a population of cytokine-dependent KLRG1(hi) CD8+ T cells is temporarily retained for improved immunity. Consideration of these findings may aid in the development of immunotherapeutic strategies against infectious disease and cancer.
Assuntos
Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Interleucina-15/farmacologia , Interleucina-7/farmacologia , Transferência Adotiva , Animais , Linfócitos T CD8-Positivos/citologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Interleucina-15/metabolismo , Interleucina-7/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ovalbumina/imunologia , Receptores de Interleucina-15/metabolismo , Receptores de Interleucina-2/metabolismo , Proteínas Recombinantes/farmacologia , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologiaRESUMO
The immune system must maintain a broad repertoire of naïve T cells in order to respond to the diverse range of pathogens that it will encounter over the course of a lifetime. Although it is known that contact with IL-7 is crucial for the survival of naïve T cells, the precise intracellular signals that mediate its effects remain obscure. An article in this issue of the European Journal of Immunology has found that IL-7 requires the coordinated action of multiple pathways to maintain naïve CD4(+) T cells.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Homeostase/imunologia , Imunidade Inata/imunologia , Animais , Citocinas/imunologiaRESUMO
Survival and intermittent proliferation of memory CD4(+) and CD8(+) T cells appear to be controlled by different homeostatic mechanisms. In particular, contact with interleukin (IL)-15 has a decisive influence on memory CD8(+) cells, but not memory CD4(+) cells. Past studies of memory CD4(+) cells have relied heavily on the use of naturally occurring memory phenotype (MP) cells as a surrogate for antigen (Ag)-specific memory cells. However, we show here that MP CD4(+) cells contain a prominent subset of rapidly proliferating major histocompatibility complex (MHC) II-dependent cells. In contrast, Ag-specific memory CD4 cells have a slow turnover rate and are MHC II independent. In irradiated hosts, these latter cells ignore IL-15 and expand in response to the elevated levels of IL-7 in the lymphopenic hosts. In contrast, in normal nonlymphopenic hosts where IL-7 levels are low, memory CD4 cells are heavily dependent on IL-15. Significantly, memory CD4(+) responsiveness to endogenous IL-15 reflects marked competition from other cells, especially CD8(+) and natural killer cells, and increases considerably after removal of these cells. Therefore, under normal physiological conditions, homeostasis of CD8(+) and CD4(+) memory cells is quite similar and involves IL-15 and IL-7.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Memória Imunológica/imunologia , Interleucina-15/imunologia , Vírus da Coriomeningite Linfocítica/imunologia , Animais , Antígenos/imunologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Homeostase , Interleucina-15/metabolismo , Interleucina-7/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Camundongos , FenótipoRESUMO
Homeostasis of T cells can be defined as the ability of the immune system to maintain normal T-cell counts and to restore T-cell numbers following T-cell depletion or expansion. These processes are governed by extrinsic signals, most notably cytokines. Two members of the common gamma chain family of cytokines, interleukin (IL)-7 and IL-15, are central to homeostatic proliferation and survival of mature CD4(+) and CD8(+) T cells. Recent evidence suggests that other cytokines, including IL-2, IL-10, IL-12, interferons and TGF-beta, as well as the transcription factors T-bet and eomesodermin all play important but different roles at distinct stages of T-cell homeostasis.
Assuntos
Citocinas/imunologia , Homeostase/imunologia , Transdução de Sinais/imunologia , Linfócitos T/imunologia , Animais , Proliferação de Células , HumanosRESUMO
The pool of memory T cells is regulated by homeostatic mechanisms to persist for prolonged periods at a relatively steady overall size. Recent work has shown that two members of the common gamma chain (gammac) family of cytokines, interleukin-7 (IL-7) and IL-15, govern homeostasis of memory T cells. These two cytokines work in conjunction to support memory T-cell survival and intermittent background proliferation. Normal animals contain significant numbers of spontaneously arising memory-phenotype (MP) cells, though whether these cells are representative of true antigen-specific memory T cells is unclear. Nevertheless, it appears that the two types of memory cells do not display identical homeostatic requirements. For antigen-specific memory CD8+ T cells, IL-7 is primarily important for survival while IL-15 is crucial for their background proliferation. For memory CD4+ T cells, IL-7 has an important role, whereas the influence of IL-15 is still unclear.
Assuntos
Memória Imunológica/imunologia , Linfócitos T/imunologia , Animais , Linfócitos T CD8-Positivos/imunologia , Homeostase/imunologia , Humanos , Interferons/imunologia , Interleucina-15/imunologia , Interleucina-7/imunologia , Camundongos , Receptores de Interleucina-2/imunologiaRESUMO
IL-15 is normally presented in vivo as a cell-associated cytokine bound to IL-15Ralpha. We show here that the biological activity of soluble IL-15 is much improved after interaction with recombinant soluble IL-15Ralpha; after injection, soluble IL-15/IL-15Ralpha complexes rapidly induce strong and selective expansion of memory-phenotype CD8(+) cells and natural killer cells. These findings imply that binding of IL-15Ralpha to IL-15 may create a conformational change that potentiates IL-15 recognition by the betagamma(c) receptor on T cells. The enhancing effect of IL-15Ralpha binding may explain why IL-15 normally functions as a cell-associated cytokine. Significantly, the results with IL-2, a soluble cytokine, are quite different; thus, IL-2 function is markedly inhibited by binding to soluble IL-2Ralpha.
Assuntos
Interleucina-15/metabolismo , Isoformas de Proteínas/metabolismo , Receptores de Interleucina-2/metabolismo , Animais , Linfócitos T CD8-Positivos/metabolismo , Proliferação de Células , Humanos , Interleucina-15/genética , Interleucina-2/genética , Interleucina-2/metabolismo , Subunidade alfa de Receptor de Interleucina-2 , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ligação Proteica , Isoformas de Proteínas/genética , Receptores de Interleucina-15 , Receptores de Interleucina-2/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
Glucocorticoid (GC) hormones cause pronounced T cell apoptosis, particularly in immature thymic T cells. This is possibly due to tissue-specific regulation of the glucocorticoid receptor (GR) gene. In mice the GR gene is transcribed from five separate promoters designated: 1A, 1B, 1C, 1D, and 1E. Nearly all cells express GR from promoters 1B-1E, but the activity of the 1A promoter has only been reported in the whole thymus or lymphocyte cell lines. To directly assess the role of GR promoter use in sensitivity to glucocorticoid-induced cell death, we have compared the activity of the GR 1A promoter with GC sensitivity in different mouse lymphocyte populations. We report that GR 1A promoter activity is restricted to thymocyte and peripheral lymphocyte populations and the cortex of the brain. The relative level of expression of the 1A promoter to the 1B-1E promoters within a lymphocyte population was found to directly correlate with susceptibility to GC-induced cell death, with the extremely GC-sensitive CD4+CD8+ thymocytes having the highest levels of GR 1A promoter activity, and the relatively GC-resistant alphabetaTCR+CD24(int/low) thymocytes and peripheral T cells having the lowest levels. DNA sequencing of the mouse GR 1A promoter revealed a putative glucocorticoid-response element. Furthermore, GR 1A promoter use and GR protein levels were increased by GC treatment in thymocytes, but not in splenocytes. These data suggest that tissue-specific differences in GR promoter use determine T cell sensitivity to glucocorticoid-induced cell death.
Assuntos
Glucocorticoides/fisiologia , Regiões Promotoras Genéticas/imunologia , Receptores de Glucocorticoides/biossíntese , Receptores de Glucocorticoides/genética , Linfócitos T/citologia , Linfócitos T/imunologia , Animais , Sequência de Bases , Morte Celular/efeitos dos fármacos , Morte Celular/genética , Morte Celular/imunologia , Diferenciação Celular/imunologia , Sobrevivência Celular/imunologia , Células Cultivadas , Córtex Cerebral/imunologia , Córtex Cerebral/metabolismo , Dexametasona/metabolismo , Dexametasona/farmacologia , Relação Dose-Resposta Imunológica , Glucocorticoides/metabolismo , Camundongos , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Especificidade de Órgãos/imunologia , RNA Mensageiro/biossíntese , Elementos de Resposta/imunologia , Análise de Sequência de DNA , Baço/citologia , Baço/imunologia , Baço/metabolismo , Linfócitos T/metabolismo , Regulação para Cima/genética , Regulação para Cima/imunologiaRESUMO
T lymphocytes (pivotal in many inflammatory pathologies) are targets for glucocorticoid hormone (GC). How TCR-mediated activation and GC signaling via glucocorticoid receptor (GR) impact on T-cell fates is not fully defined. We delineated here the expression of a recently identified glucocorticoid-induced TNF receptor (GITR) induced by GC and by TCR-mediated T-cell activation in GC receptor (GR)-deficient mice (GR-/-). We also compared the action of GC on GITR+ and GITR- T cells by monitoring apoptosis, proliferation and cytokine production stimulated by anti-CD3 antibody. By using GR-/- mice, we observed that the development of GITR+ T cells (both in thymus and periphery) is not dependent upon GR signaling. This contradicts the implication of GITR's name reflecting GC induction. TCR-mediated T-cell activation induced GITR expression in both GR+/+ and GR-/- cells. Somewhat unexpectedly, there was very modest GITR upregulation on GR+/+ T cells by a range of GC doses (10(-8) to 10(-6) M). Constitutive expression of GITR by a subset of CD4+ cells did not significantly render them resistant to GC-induced cell death. However, TCR-induced GITR upregulation on GR+/+ T cells was correlated with resistance to GC-mediated apoptosis suggesting that GITR, in conjunction with other (as yet unidentified) TCR-induced factors, protects T cells from apoptosis. Thus, even though GC is a potent inducer of apoptosis of T cells, activated T cells are resistant to GC-mediated killing. Meanwhile, although GC suppressed anti-CD3-induced cytokine production, cell proliferation was unaffected by GC in GR+/+ mice. GR deficiency has no effect on anti-CD3-induced cytokine production and proliferation. Our findings also have implications for GC treatment in that it would be more difficult to abrogate an ongoing T-cell mediated inflammatory response than to prevent its induction.
Assuntos
Apoptose/efeitos dos fármacos , Ativação Linfocitária , Receptores de Antígenos de Linfócitos T/fisiologia , Receptores de Fator de Crescimento Neural/biossíntese , Receptores do Fator de Necrose Tumoral/biossíntese , Linfócitos T/imunologia , Animais , Complexo CD3/imunologia , Citocinas/biossíntese , Dexametasona/farmacologia , Proteína Relacionada a TNFR Induzida por Glucocorticoide , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Glucocorticoides/fisiologiaRESUMO
To determine the tissue-specific functions of SOCS-1, mice were generated in which the SOCS-1 gene could be deleted in individual tissues. A reporter gene of SOCS-1 promoter activity was also inserted. Using the reporter, high SOCS-1 expression was found at the CD4(+)CD8(+) stage in thymocyte development. To investigate the function of this expression, the SOCS-1 gene was specifically deleted throughout the thymocyte/T/NKT cell compartment. Unlike SOCS-1(-/-) mice, these mice did not develop lethal multiorgan inflammation but developed multiple lymphoid abnormalities, including enhanced differentiation of thymocytes toward CD8(+) T cells and very high percentages of peripheral CD8(+) T cells with a memory phenotype (CD44(hi)CD25(lo)CD69(lo)). These phenotypes were found to correlate with hypersensitivity to the gamma-common family of cytokines.
Assuntos
Linfócitos T CD8-Positivos/fisiologia , Proteínas de Transporte/fisiologia , Interleucina-7/farmacologia , Proteínas Repressoras , Animais , Proteínas de Transporte/genética , Diferenciação Celular , Memória Imunológica , Imunofenotipagem , Integrases/genética , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/análise , Proteína 1 Supressora da Sinalização de Citocina , Proteínas Supressoras da Sinalização de Citocina , Proteínas Virais/genéticaRESUMO
Suppressor of cytokine signaling (SOCS)-1 is a member of a family of proteins that negatively regulate cytokine signaling pathways. We have previously established that SOCS-1 is a key regulator of IFN-gamma signaling and that IFN-gamma is responsible for the complex inflammatory disease that leads to the death of SOCS-1-deficient mice. In this study, we provide evidence that SOCS-1 is also a critical regulator of IFN-gamma-independent immunoregulatory factors. Mice lacking both SOCS-1 and IFN-gamma, although outwardly healthy, have clear abnormalities in their immune system, including a reduced ratio of CD4:CD8 T cells in lymphoid tissues and increased expression of T cell activation markers. To examine the contribution of TCR Ag specificity to these immune defects, we have generated two lines of SOCS-1-deficient mice expressing a transgenic TCR specific for an exogenous Ag, OVA (OT-I and OT-II). Although TCR transgenic SOCS-1(-/-) mice have a longer lifespan than nontransgenic SOCS-1(-/-) mice, they still die as young adults with inflammatory disease and the TCR transgenic SOCS-1(-/-) T cells appear activated despite the absence of OVA. This suggests that both Ag-dependent and -independent mechanisms contribute to the disease in SOCS-1-deficient mice. Thus, SOCS-1 is a critical regulator of T cell activation and homeostasis, and its influence extends beyond regulating IFN-gamma signaling.
Assuntos
Proteínas de Transporte/fisiologia , Citocinas/antagonistas & inibidores , Citocinas/fisiologia , Homeostase/imunologia , Interferon gama/fisiologia , Proteínas Repressoras , Transdução de Sinais/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Relação CD4-CD8 , Proteínas de Transporte/genética , Epitopos de Linfócito T/imunologia , Feto , Homeostase/genética , Imunofenotipagem , Interferon gama/deficiência , Interferon gama/genética , Doenças Linfáticas/genética , Doenças Linfáticas/imunologia , Doenças Linfáticas/patologia , Ativação Linfocitária/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Técnicas de Cultura de Órgãos , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de Sinais/genética , Proteína 1 Supressora da Sinalização de Citocina , Proteínas Supressoras da Sinalização de Citocina , Subpopulações de Linfócitos T/metabolismo , Timo/citologia , Timo/imunologiaRESUMO
Peripheral activation can cause bystander thymocyte death by eliciting a "cytokine storm." This event complicates in vivo studies using exogenous ligand-induced models of negative selection. A stable transgenic model that selectively eliminates peripheral CD4 cells has allowed us to analyze negative selection as direct cognate events in two T cell receptor transgenic mice, OT-II and DO11. Whereas cognate peptide induced a massive deletion in double-positive (DP) cells in mice with peripheral CD4 cells, this DP deletion was modest in mice lacking peripheral CD4 cells. Using BrdUrd and annexin V staining, we found that negative selection primarily occurs in a cohort of DP cells and the absence of single-positive (SP) cells is largely caused by reduction in the cohort of DP precursors. Moreover, the fates of DP cells and SP cells after antigen exposure were vastly different. Whereas SP cells up-regulated uniformly their CD69 and CD44 levels, increased their cell size, and survived after antigen exposure, DP cells had less CD69 and CD44 up-regulation, no size change, and promptly died. Thus, negative selection represents an "abortive" activation different from activation-induced cell death of mature T cells.
Assuntos
Deleção Clonal , Timo/metabolismo , Animais , Anexina A5/farmacologia , Antígenos/metabolismo , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos T/biossíntese , Apoptose , Bromodesoxiuridina/farmacologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Corantes/farmacologia , Ativação Enzimática , Citometria de Fluxo , Receptores de Hialuronatos/biossíntese , Imunofenotipagem , Lectinas Tipo C , Ligantes , Ativação Linfocitária , Camundongos , Camundongos Transgênicos , Peptídeos/química , Baço/citologia , Timo/citologia , Regulação para CimaRESUMO
The involvement of glucocorticoid receptor (GR) signaling in T cell development is highly controversial, with several studies for and against. We have previously demonstrated that GR(-/-) mice, which usually die at birth because of impaired lung development, exhibit normal T cell development, at least in embryonic mice and in fetal thymus organ cultures. To directly investigate the role of GR signaling in adult T cell development, we analyzed the few GR(-/-) mice that occasionally survive birth, and irradiated mice reconstituted with GR(-/-) fetal liver precursors. All thymic and peripheral T cells, as well as other leukocyte lineages, developed and were maintained at normal levels. Anti-CD3-induced cell death of thymocytes in vitro, T cell repertoire heterogeneity and T cell proliferation in response to anti-CD3 stimulation were normal in the absence of GR signaling. Finally, we show that metyrapone, an inhibitor of glucocorticoid synthesis (commonly used to demonstrate a role for glucocorticoids in T cell development), impaired thymocyte development regardless of GR genotype indicating that this reagent inhibits thymocyte development in a glucocorticoid-independent fashion. These data demonstrate that GR signaling is not required for either normal T cell development or peripheral maintenance in embryonic or adult mice.
Assuntos
Receptores de Glucocorticoides/deficiência , Linfócitos T/citologia , Linfócitos T/metabolismo , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Diferenciação Celular/efeitos dos fármacos , Transplante de Tecido Fetal , Hepatócitos/citologia , Hepatócitos/transplante , Técnicas In Vitro , Metirapona/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Quimera por Radiação , Receptores de Antígenos de Linfócitos T/metabolismo , Receptores de Glucocorticoides/genética , Transdução de Sinais , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Timo/citologia , Timo/imunologia , Timo/metabolismoRESUMO
During lymphocyte development, the assembly of genes coding for antigen receptors occurs by the combinatorial linking of gene segments. The stochastic nature of this process gives rise to lymphocytes that can recognize self-antigens, thereby having the potential to induce autoimmune disease. Such autoreactive lymphocytes can be silenced by developmental arrest or unresponsiveness (anergy), or can be deleted from the repertoire by cell death. In the thymus, developing T lymphocytes (thymocytes) bearing a T-cell receptor (TCR)-CD3 complex that engages self-antigens are induced to undergo programmed cell death (apoptosis), but the mechanisms ensuring this 'negative selection' are unclear. We now report that thymocytes lacking the pro-apoptotic Bcl-2 family member Bim (also known as Bcl2l11) are refractory to apoptosis induced by TCR-CD3 stimulation. Moreover, in transgenic mice expressing autoreactive TCRs that provoke widespread deletion, Bim deficiency severely impaired thymocyte killing. TCR ligation upregulated Bim expression and promoted interaction of Bim with Bcl-XL, inhibiting its survival function. These findings identify Bim as an essential initiator of apoptosis in thymocyte-negative selection.