Loss of BIM in T cells results in BCL-2 family BH3-member compensation but incomplete cell death sensitivity normalization


BIM is the master BH3-only BCL-2 family regulator of lymphocyte survival. To understand how long-term loss of BIM affects apoptotic resistance in T cells we studied animals with T cell-specific deletion of Bim. Unlike CD19CREBimfl/fl animals, LCKCREBimfl/fl mice have pronounced early lymphocytosis followed by normalization of lymphocyte counts over time. This normalization occurred in mature T cells, as thymocyte development and apoptotic sensitivity remained abnormal in LCKCREBimfl/fl mice. T cells from aged mice experienced normalization of their absolute cell numbers and responses against various apoptotic stimuli. mRNA expression levels of BCL-2 family proteins in CD4+ and CD8+ T cells from young and old mice revealed upregulation of several BH3-only proteins, including Puma, Noxa, and Bmf. Despite upregulation of various BH3 proteins, there were no differences in anti-apoptotic BCL-2 protein dependency in these cells. However, T cells had continued resistance to direct BIM BH3-induced mitochondrial depolarization. This study further highlights the importance of BIM in cell death maintenance in T cells and provides new insight into the dynamism underlying BH3-only regulation of T cell homeostasis versus induced cell death and suggests that CD4+ and CD8+ T cells compensate differently in response to loss of Bim.

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  1. 1.

    Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26(4):239–257

    CAS  Article  Google Scholar 

  2. 2.

    Ashkenazi A, Dixit VM (1998) Death receptors: signaling and modulation. Science 281(5381):1305–1308

    CAS  Article  Google Scholar 

  3. 3.

    Czabotar PE, Lessene G, Strasser A, Adams JM (2014) Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol 15(1):49–63. https://doi.org/10.1038/nrm3722

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116(2):205–219

    CAS  Article  Google Scholar 

  5. 5.

    Ludwig LM, Nassin ML, Hadji A, LaBelle JL (2016) Killing two cells with one stone: pharmacologic BCL-2 family targeting for cancer cell death and immune modulation. Front Pediatr 4:135. https://doi.org/10.3389/fped.2016.00135

    Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Shamas-Din A, Brahmbhatt H, Leber B, Andrews DW (2011) BH3-only proteins: orchestrators of apoptosis. Biochim Biophys Acta 1813(4):508–520. https://doi.org/10.1016/j.bbamcr.2010.11.024

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Chen L, Willis SN, Wei A, Smith BJ, Fletcher JI, Hinds MG, Colman PM, Day CL, Adams JM, Huang DC (2005) Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell 17(3):393–403. https://doi.org/10.1016/j.molcel.2004.12.030

    CAS  Article  Google Scholar 

  8. 8.

    Chen HC, Kanai M, Inoue-Yamauchi A, Tu HC, Huang Y, Ren D, Kim H, Takeda S, Reyna DE, Chan PM, Ganesan YT, Liao CP, Gavathiotis E, Hsieh JJ, Cheng EH (2015) An interconnected hierarchical model of cell death regulation by the BCL-2 family. Nat Cell Biol 17(10):1270–1281. https://doi.org/10.1038/ncb3236

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR, Newmeyer DD (2005) BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol Cell 17(4):525–535. https://doi.org/10.1016/j.molcel.2005.02.003

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Ren D, Tu HC, Kim H, Wang GX, Bean GR, Takeuchi O, Jeffers JR, Zambetti GP, Hsieh JJ, Cheng EH (2010) BID, BIM, and PUMA are essential for activation of the BAX- and BAK-dependent cell death program. Science 330(6009):1390–1393. https://doi.org/10.1126/science.1190217

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Bouillet P, Metcalf D, Huang DC, Tarlinton DM, Kay TW, Kontgen F, Adams JM, Strasser A (1999) Proapoptotic Bcl-2 relative Bim required for certain apoptotic responses, leukocyte homeostasis, and to preclude autoimmunity. Science 286(5445):1735–1738

    CAS  Article  Google Scholar 

  12. 12.

    Bouillet P, Purton JF, Godfrey DI, Zhang LC, Coultas L, Puthalakath H, Pellegrini M, Cory S, Adams JM, Strasser A (2002) BH3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes. Nature 415(6874):922–926. https://doi.org/10.1038/415922a

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Enders A, Bouillet P, Puthalakath H, Xu Y, Tarlinton DM, Strasser A (2003) Loss of the pro-apoptotic BH3-only Bcl-2 family member Bim inhibits BCR stimulation-induced apoptosis and deletion of autoreactive B cells. J Exp Med 198(7):1119–1126. https://doi.org/10.1084/jem.20030411

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Hernandez JB, Newton RH, Walsh CM (2010) Life and death in the thymus–cell death signaling during T cell development. Curr Opin Cell Biol 22(6):865–871. https://doi.org/10.1016/j.ceb.2010.08.003

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Renault TT, Chipuk JE (2013) Getting away with murder: how does the BCL-2 family of proteins kill with immunity? Ann N Y Acad Sci 1285:59–79. https://doi.org/10.1111/nyas.12045

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Sionov RV, Vlahopoulos SA, Granot Z (2015) Regulation of Bim in health and disease. Oncotarget 6(27):23058–23134. https://doi.org/10.18632/oncotarget.5492

    Article  PubMed  PubMed Central  Google Scholar 

  17. 17.

    Strasser A (2005) The role of BH3-only proteins in the immune system. Nat Rev Immunol 5(3):189–200. https://doi.org/10.1038/nri1568

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Herold MJ, Stuchbery R, Merino D, Willson T, Strasser A, Hildeman D, Bouillet P (2014) Impact of conditional deletion of the pro-apoptotic BCL-2 family member BIM in mice. Cell Death Dis 5:e1446. https://doi.org/10.1038/cddis.2014.409

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Hildeman D, Jorgensen T, Kappler J, Marrack P (2007) Apoptosis and the homeostatic control of immune responses. Curr Opin Immunol 19(5):516–521. https://doi.org/10.1016/j.coi.2007.05.005

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. 20.

    Shukla S, Saxena S, Singh BK, Kakkar P (2017) BH3-only protein BIM: an emerging target in chemotherapy. Eur J Cell Biol 96(8):728–738. https://doi.org/10.1016/j.ejcb.2017.09.002

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Carrington EM, Zhan Y, Brady JL, Zhang JG, Sutherland RM, Anstee NS, Schenk RL, Vikstrom IB, Delconte RB, Segal D, Huntington ND, Bouillet P, Tarlinton DM, Huang DC, Strasser A, Cory S, Herold MJ, Lew AM (2017) Anti-apoptotic proteins BCL-2, MCL-1 and A1 summate collectively to maintain survival of immune cell populations both in vitro and in vivo. Cell Death Differ 24(5):878–888. https://doi.org/10.1038/cdd.2017.30

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Biswas S, Shi Q, Matise L, Cleveland S, Dave U, Zinkel S (2010) A role for proapoptotic Bax and Bak in T-cell differentiation and transformation. Blood 116(24):5237–5246. https://doi.org/10.1182/blood-2010-04-279687

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Katz SG, Labelle JL, Meng H, Valeriano RP, Fisher JK, Sun H, Rodig SJ, Kleinstein SH, Walensky LD (2014) Mantle cell lymphoma in cyclin D1 transgenic mice with Bim-deficient B cells. Blood 123(6):884–893. https://doi.org/10.1182/blood-2013-04-499079

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Takeuchi O, Fisher J, Suh H, Harada H, Malynn BA, Korsmeyer SJ (2005) Essential role of BAX,BAK in B cell homeostasis and prevention of autoimmune disease. Proc Natl Acad Sci USA 102(32):11272–11277. https://doi.org/10.1073/pnas.0504783102

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Opferman JT, Letai A, Beard C, Sorcinelli MD, Ong CC, Korsmeyer SJ (2003) Development and maintenance of B and T lymphocytes requires antiapoptotic MCL-1. Nature 426(6967):671–676. https://doi.org/10.1038/nature02067

    CAS  Article  Google Scholar 

  26. 26.

    LaBelle JL, Hanke CA, Blazar BR, Truitt RL (2002) Negative effect of CTLA-4 on induction of T-cell immunity in vivo to B7-1+, but not B7-2+, murine myelogenous leukemia. Blood 99(6):2146–2153

    CAS  Article  Google Scholar 

  27. 27.

    Pitter K, Bernal F, Labelle J, Walensky LD (2008) Dissection of the BCL-2 family signaling network with stabilized alpha-helices of BCL-2 domains. Methods Enzymol 446:387–408. https://doi.org/10.1016/S0076-6879(08)01623-6

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Murase K, Kim HT, Bascug OR, Kawano Y, Ryan J, Matsuoka K, Davids MS, Koreth J, Ho VT, Cutler C, Armand P, Alyea EP, Blazar BR, Antin JH, Soiffer RJ, Letai A, Ritz J (2014) Increased mitochondrial apoptotic priming of human regulatory T cells after allogeneic hematopoietic stem cell transplantation. Haematologica 99(9):1499–1508. https://doi.org/10.3324/haematol.2014.104166

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Ryan J, Letai A (2013) BH3 profiling in whole cells by fluorimeter or FACS. Methods 61(2):156–164. https://doi.org/10.1016/j.ymeth.2013.04.006

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Hutcheson J, Perlman H (2007) Loss of Bim results in abnormal accumulation of mature CD4-CD8-CD44-CD25- thymocytes. Immunobiology 212(8):629–636. https://doi.org/10.1016/j.imbio.2007.05.003

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Li L, Hsu HC, Grizzle WE, Stockard CR, Ho KJ, Lott P, Yang PA, Zhang HG, Mountz JD (2003) Cellular mechanism of thymic involution. Scand J Immunol 57(5):410–422

    CAS  Article  Google Scholar 

  32. 32.

    Erlacher M, Labi V, Manzl C, Bock G, Tzankov A, Hacker G, Michalak E, Strasser A, Villunger A (2006) Puma cooperates with Bim, the rate-limiting BH3-only protein in cell death during lymphocyte development, in apoptosis induction. J Exp Med 203(13):2939–2951. https://doi.org/10.1084/jem.20061552

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. 33.

    Chougnet CA, Tripathi P, Lages CS, Raynor J, Sholl A, Fink P, Plas DR, Hildeman DA (2011) A major role for Bim in regulatory T cell homeostasis. J Immunol 186(1):156–163. https://doi.org/10.4049/jimmunol.1001505

    CAS  Article  PubMed  Google Scholar 

  34. 34.

    Harpaz I, Bhattacharya U, Elyahu Y, Strominger I, Monsonego A (2017) Old mice accumulate activated effector CD4 T cells refractory to regulatory T cell-induced immunosuppression. Front Immunol 8:283. https://doi.org/10.3389/fimmu.2017.00283

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Tischner D, Gaggl I, Peschel I, Kaufmann M, Tuzlak S, Drach M, Thuille N, Villunger A, Jan Wiegers G (2012) Defective cell death signalling along the Bcl-2 regulated apoptosis pathway compromises Treg cell development and limits their functionality in mice. J Autoimmun 38(1):59–69. https://doi.org/10.1016/j.jaut.2011.12.008

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    McCaughtry TM, Wilken MS, Hogquist KA (2007) Thymic emigration revisited. J Exp Med 204(11):2513–2520. https://doi.org/10.1084/jem.20070601

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Yates AJ (2014) Theories and quantification of thymic selection. Front Immunol 5:13. https://doi.org/10.3389/fimmu.2014.00013

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Happo L, Strasser A, Cory S (2012) BH3-only proteins in apoptosis at a glance. J Cell Sci 125(Pt 5):1081–1087. https://doi.org/10.1242/jcs.090514

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  39. 39.

    Certo M, Del Gaizo Moore V, Nishino M, Wei G, Korsmeyer S, Armstrong SA, Letai A (2006) Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell 9(5):351–365. https://doi.org/10.1016/j.ccr.2006.03.027

    CAS  Article  PubMed  Google Scholar 

  40. 40.

    Deng J, Carlson N, Takeyama K, Dal Cin P, Shipp M, Letai A (2007) BH3 profiling identifies three distinct classes of apoptotic blocks to predict response to ABT-737 and conventional chemotherapeutic agents. Cancer Cell 12(2):171–185. https://doi.org/10.1016/j.ccr.2007.07.001

    CAS  Article  PubMed  Google Scholar 

  41. 41.

    Carrington EM, Tarlinton DM, Gray DH, Huntington ND, Zhan Y, Lew AM (2017) The life and death of immune cell types: the role of BCL-2 anti-apoptotic molecules. Immunol Cell Biol 95(10):870–877. https://doi.org/10.1038/icb.2017.72

    CAS  Article  PubMed  Google Scholar 

  42. 42.

    Merino D, Giam M, Hughes PD, Siggs OM, Heger K, O’Reilly LA, Adams JM, Strasser A, Lee EF, Fairlie WD, Bouillet P (2009) The role of BH3-only protein Bim extends beyond inhibiting Bcl-2-like prosurvival proteins. J Cell Biol 186(3):355–362. https://doi.org/10.1083/jcb.200905153

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  43. 43.

    Wojciechowski S, Tripathi P, Bourdeau T, Acero L, Grimes HL, Katz JD, Finkelman FD, Hildeman DA (2007) Bim/Bcl-2 balance is critical for maintaining naive and memory T cell homeostasis. J Exp Med 204(7):1665–1675. https://doi.org/10.1084/jem.20070618

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Lages CS, Suffia I, Velilla PA, Huang B, Warshaw G, Hildeman DA, Belkaid Y, Chougnet C (2008) Functional regulatory T cells accumulate in aged hosts and promote chronic infectious disease reactivation. J Immunol 181(3):1835–1848

    CAS  Article  Google Scholar 

  45. 45.

    Leverson JD, Phillips DC, Mitten MJ, Boghaert ER, Diaz D, Tahir SK, Belmont LD, Nimmer P, Xiao Y, Ma XM, Lowes KN, Kovar P, Chen J, Jin S, Smith M, Xue J, Zhang H, Oleksijew A, Magoc TJ, Vaidya KS, Albert DH, Tarrant JM, La N, Wang L, Tao ZF, Wendt MD, Sampath D, Rosenberg SH, Tse C, Huang DC, Fairbrother WJ, Elmore SW, Souers AJ (2015) Exploiting selective BCL-2 family inhibitors to dissect cell survival dependencies and define improved strategies for cancer therapy. Sci Transl Med 7(279):279ra240. https://doi.org/10.1126/scitranslmed.aaa4642

    CAS  Article  Google Scholar 

  46. 46.

    Renault TT, Elkholi R, Bharti A, Chipuk JE (2014) B cell lymphoma-2 (BCL-2) homology domain 3 (BH3) mimetics demonstrate differential activities dependent upon the functional repertoire of pro-and anti-apoptotic BCL-2 family proteins. J Biol Chem 289(38):26481–26491. https://doi.org/10.1074/jbc.M114.569632

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  47. 47.

    Lopez J, Bessou M, Riley JS, Giampazolias E, Todt F, Rochegue T, Oberst A, Green DR, Edlich F, Ichim G, Tait SW (2016) Mito-priming as a method to engineer Bcl-2 addiction. Nat Commun 7:10538. https://doi.org/10.1038/ncomms10538

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  48. 48.

    Letai A, Bassik MC, Walensky LD, Sorcinelli MD, Weiler S, Korsmeyer SJ (2002) Distinct BH3 domains either sensitize or activate mitochondrial apoptosis, serving as prototype cancer therapeutics. Cancer Cell 2(3):183–192

    CAS  Article  Google Scholar 

  49. 49.

    Delgado M, Tesfaigzi Y (2013) BH3-only proteins, Bmf and Bim, in autophagy. Cell Cycle 12(22):3453–3454. https://doi.org/10.4161/cc.26696

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  50. 50.

    Labi V, Erlacher M, Kiessling S, Manzl C, Frenzel A, O’Reilly L, Strasser A, Villunger A (2008) Loss of the BH3-only protein Bmf impairs B cell homeostasis and accelerates gamma irradiation-induced thymic lymphoma development. J Exp Med 205(3):641–655. https://doi.org/10.1084/jem.20071658

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  51. 51.

    Labi V, Woess C, Tuzlak S, Erlacher M, Bouillet P, Strasser A, Tzankov A, Villunger A (2014) Deregulated cell death and lymphocyte homeostasis cause premature lethality in mice lacking the BH3-only proteins Bim and Bmf. Blood 123(17):2652–2662. https://doi.org/10.1182/blood-2013-11-537217

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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This work was supported by research funds from the National Institutes of Health: NCI K08-CA151450 (to J.L.L.) and the Comer Children’s Hospital Development Board, University of Chicago (to J.L.L). We would like to thank Loren Walensky for initial critical review of our manuscript.

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Ludwig, L.M., Roach, L.E., Katz, S.G. et al. Loss of BIM in T cells results in BCL-2 family BH3-member compensation but incomplete cell death sensitivity normalization. Apoptosis 25, 247–260 (2020). https://doi.org/10.1007/s10495-020-01593-6

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  • BCL-2
  • T cells
  • BIM
  • Apoptosis