Abstract
T cells developing in the thymus undergo rigorous positive and negative selection to ensure that those exported to peripheral lymphoid organs bear T-cell receptors (TCRs) capable of reacting with foreign antigens but tolerant of self. At each checkpoint, whether a thymocyte survives or dies is determined by antiapoptotic and proapoptotic Bcl-2 family members. We used Mcl-1 transgenic (tg) mice to investigate the impact of elevated expression of antiapoptotic Mcl-1 on thymocyte apoptosis and selection, making a side-by-side comparison with thymocytes from BCL-2tg mice. Mcl-1 was as effective as Bcl-2 at protecting thymocytes against spontaneous cell death, diverse cytotoxic insults and TCR–CD3 stimulation-driven apoptosis. In three different TCR tg models, Mcl-1 markedly enhanced positive selection of thymocytes, as did Bcl-2. In H-Y TCR tg mice, elevated Mcl-1 and Bcl-2 were equally effective at inhibiting deletion of autoreactive thymocytes. However, in the OT-1tg model where deletion is mediated by a peripheral antigen whose expression is regulated by Aire, Mcl-1 was less effective than Bcl-2. Thus, the capacity of Mcl-1 overexpression to inhibit apoptosis triggered by TCR stimulation apparently depends on the thymocyte subset subject to deletion, presumably due to differences in the profiles of proapoptotic Bcl-2 family members mediating the deletion.
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Abbreviations
- TCR:
-
T-cell receptor
- DN:
-
double-negative
- DP:
-
double-positive
- SP:
-
single-positive
- MHC:
-
major histocompatibility complex
- PMA:
-
phorbol 12-myristate 13-acetate
- WT:
-
wild-type
- BM:
-
bone marrow
- tg:
-
transgenic
- OVA:
-
ovalbumin
- RIP:
-
rat insulin promoter
- WEHI:
-
The Walter and Eliza Hall Institute
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Acknowledgements
We thank our colleagues C Vandenberg, C Scott, K Heger and P Bouillet for useful discussions; L O’Reilly and D Huang for reagents; J Blyth and F Kupresanin for assistance; B Helbert and C Young for genotyping; K McKenzie, T Camilleri and G Siciliano for mouse husbandry; and WEHI cytometry and histology services. This work was supported by postdoctoral fellowships from EMBO and the Human Frontier in Science Program (KJC); NHMRC Career Development Fellowship (DHDG) (CDF1 #637353); NHMRC Australia Fellowship (AS); research grants from National Health and Medical Research Council (NHMRC) (programme grant 461221) and National Cancer Institute (CA43540); and operational infrastructure grants through the Australian Government IRISS and the Victorian State Government OIS.
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Campbell, K., Gray, D., Anstee, N. et al. Elevated Mcl-1 inhibits thymocyte apoptosis and alters thymic selection. Cell Death Differ 19, 1962–1971 (2012). https://doi.org/10.1038/cdd.2012.84
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DOI: https://doi.org/10.1038/cdd.2012.84
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