Maintenance of the HIV reservoir is antagonized by selective BCL2 inhibition

Journal of Virology

Volumn 91, Issue 11, 2017, Pages

  Cummins, Nathan W ^a   Sainski Nguyen, Amy M ^a   Natesampillai, Sekar ^a   Aboulnasr, Fatma ^a   Kaufmann, Scott ^a   Badley, Andrew D ^a  

^a MAYO CLINIC   (United States)

Author keywords

Antiapoptosis; Bcl 2 family; HIV; Reservoir

Indexed keywords

DIMETHYL SULFOXIDE; INTERLEUKIN 7; PROTEIN BCL 2; VENETOCLAX; FUSED HETEROCYCLIC RINGS; SULFONAMIDE;

ARTICLE; CD4+ T LYMPHOCYTE; CELL DEATH; CELL KILLING; CELL PROLIFERATION; CONTROLLED STUDY; DRUG EFFECT; DRUG MECHANISM; HOST CELL; HUMAN; HUMAN CELL; HUMAN IMMUNODEFICIENCY VIRUS; IMMUNOSTIMULATION; NONHUMAN; PERSISTENT INFECTION; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN FUNCTION; ANTAGONISTS AND INHIBITORS; APOPTOSIS; CELL CULTURE; DRUG EFFECTS; HUMAN IMMUNODEFICIENCY VIRUS 1; IMMUNOLOGY; JURKAT CELL LINE; METABOLISM; PHYSIOLOGY; VIROLOGY; VIRUS LATENCY; VIRUS REPLICATION;

APOPTOSIS; BRIDGED BICYCLO COMPOUNDS, HETEROCYCLIC; CD4-POSITIVE T-LYMPHOCYTES; CELL DEATH; CELLS, CULTURED; HIV-1; HUMANS; JURKAT CELLS; PROTO-ONCOGENE PROTEINS C-BCL-2; SULFONAMIDES; VIRUS LATENCY; VIRUS REPLICATION;

EID: 85019258735     PISSN: 0022538X     EISSN: 10985514     Source Type: Journal    
DOI: 10.1128/JVI.00012-17     Document Type: Article

References (68)

1. Chun TW, Engel D, Berrey MM, Shea T, Corey L, Fauci AS. 1998. Early establishment of a pool of latently infected, resting CD4(+) T cells during primary HIV-1 infection. Proc Natl Acad Sci U S A 95:8869-8873. https://doi.org/10.1073/pnas.95.15.8869.
2. Chavez L, Calvanese V, Verdin E. 2015. HIV latency is established directly and early in both resting and activated primary CD4 T cells. PLoS Pathog 11:e1004955. https://doi.org/10.1371/journal.ppat.1004955.
3. Siliciano JM, Siliciano RF. 2015. The remarkable stability of the latent reservoir for HIV-1 in resting memory CD4+ T cells. J Infect Dis 212: 1345-1347. https://doi.org/10.1093/infdis/jiv219.
4. von Stockenstrom S, Odevall L, Lee E, Sinclair E, Bacchetti P, Killian M, Epling L, Shao W, Hoh R, Ho T, Faria NR, Lemey P, Albert J, Hunt P, Loeb L, Pilcher C, Poole L, Hatano H, Somsouk M, Douek D, Boritz E, Deeks SG, Hecht FM, Palmer S. 2015. Longitudinal genetic characterization reveals that cell proliferation maintains a persistent HIV type 1 DNA pool during effective HIV therapy. J Infect Dis 212:596-607. https://doi.org/10.1093/infdis/jiv092.
5. Kearney MF, Wiegand A, Shao W, Coffin JM, Mellors JW, Lederman M, Gandhi RT, Keele BF, Li JZ. 2015. Origin of rebound plasma HIV includes cells with identical proviruses that are transcriptionally active before stopping of antiretroviral therapy. J Virol 90:1369-1376. https://doi.org/10.1128/JVI.02139-15.
6. Simonetti FR, Sobolewski MD, Fyne E, Shao W, Spindler J, Hattori J, Anderson EM, Watters SA, Hill S, Wu X, Wells D, Su L, Luke BT, Halvas EK, Besson G, Penrose KJ, Yang Z, Kwan RW, Van Waes C, Uldrick T, Citrin DE, Kovacs J, Polis MA, Rehm CA, Gorelick R, Piatak M, Keele BF, Kearney MF, Coffin JM, Hughes SH, Mellors JW, Maldarelli F. 2016. Clonally expanded CD4+ T cells can produce infectious HIV-1 in vivo. Proc Natl Acad Sci U S A 113:1883-1888. https://doi.org/10.1073/pnas.1522675113.
7. Chomont N, El-Far M, Ancuta P, Trautmann L, Procopio FA, Yassine-Diab B, Boucher G, Boulassel MR, Ghattas G, Brenchley JM, Schacker TW, Hill BJ, Douek DC, Routy JP, Haddad EK, Sekaly RP. 2009. HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. Nat Med 15:893-900. https://doi.org/10.1038/nm.1972.
8. Vandergeeten C, Fromentin R, DaFonseca S, Lawani MB, Sereti I, Lederman MM, Ramgopal M, Routy JP, Sekaly RP, Chomont N. 2013. Interleukin-7 promotes HIV persistence during antiretroviral therapy. Blood 121:4321-4329. https://doi.org/10.1182/blood-2012-11-465625.
9. Günthard HF, Frost SD, Leigh-Brown AJ, Ignacio CC, Kee K, Perelson AS, Spina CA, Havlir DV, Hezareh M, Looney DJ, Richman DD, Wong JK. 1999. Evolution of envelope sequences of human immunodeficiency virus type 1 in cellular reservoirs in the setting of potent antiviral therapy. J Virol 73:9404-9412.
10. Frenkel LM, Wang Y, Learn GH, McKernan JL, Ellis GM, Mohan KM, Holte SE, De Vange SM, Pawluk DM, Melvin AJ, Lewis PF, Heath LM, Beck IA, Mahalanabis M, Naugler WE, Tobin NH, Mullins JI. 2003. Multiple viral genetic analyses detect low-level human immunodeficiency virus type 1 replication during effective highly active antiretroviral therapy. J Virol 77:5721-5730. https://doi.org/10.1128/JVI.77.10.5721-5730.2003.
11. Lorenzo-Redondo R, Fryer HR, Bedford T, Kim EY, Archer J, Kosakovsky Pond SL, Chung YS, Penugonda S, Chipman JG, Fletcher CV, Schacker TW, Malim MH, Rambaut A, Haase AT, McLean AR, Wolinsky SM. 2016. Persistent HIV-1 replication maintains the tissue reservoir during therapy. Nature 530:51-56. https://doi.org/10.1038/nature16933.
12. Henrich TJ, Hanhauser E, Marty FM, Sirignano MN, Keating S, Lee TH, Robles YP, Davis BT, Li JZ, Heisey A, Hill AL, Busch MP, Armand P, Soiffer RJ, Altfeld M, Kuritzkes DR. 2014. Antiretroviral-free HIV-1 remission and viral rebound after allogeneic stem cell transplantation: report of 2 cases. Ann Intern Med 161:319-327. https://doi.org/10.7326/M14-1027.
13. Mavigner M, Watkins B, Lawson B, Lee ST, Chahroudi A, Kean L, Silvestri G. 2014. Persistence of virus reservoirs in ART-treated SHIV-infected rhesus macaques after autologous hematopoietic stem cell transplant. PLoS Pathog 10:e1004406. https://doi.org/10.1371/journal.ppat.1004406.
14. Kaminski R, Bella R, Yin C, Otte J, Ferrante P, Gendelman HE, Li H, Booze R, Gordon J, Hu W, Khalili K. 2016. Excision of HIV-1 DNA by gene editing: a proof-of-concept in vivo study. Gene Ther 23:690-695. https://doi.org/10.1038/gt.2016.41.
15. Kaminski R, Chen Y, Salkind J, Bella R, Young WB, Ferrante P, Karn J, Malcolm T, Hu W, Khalili K. 2016. Negative feedback regulation of HIV-1 by gene editing strategy. Sci Rep 6:31527. https://doi.org/10.1038/srep31527.
16. Scholler J, Brady TL, Binder-Scholl G, Hwang WT, Plesa G, Hege KM, Vogel AN, Kalos M, Riley JL, Deeks SG, Mitsuyasu RT, Bernstein WB, Aronson NE, Levine BL, Bushman FD, June CH. 2012. Decade-long safety and function of retroviral-modified chimeric antigen receptor T cells. Sci Transl Med 4:132ra53. https://doi.org/10.1126/scitranslmed.3003761.
17. Sahu GK, Sango K, Selliah N, Ma Q, Skowron G, Junghans RP. 2013. Anti-HIV designer T cells progressively eradicate a latently infected cell line by sequentially inducing HIV reactivation then killing the newly gp120-positive cells. Virology 446:268-275. https://doi.org/10.1016/j.virol.2013.08.002.
18. Liu B, Zou F, Lu L, Chen C, He D, Zhang X, Tang X, Liu C, Li L, Zhang H. 2016 Chimeric antigen receptor T cells guided by the single-chain Fv of a broadly neutralizing antibody specifically and effectively eradicate virus reactivated from latency in CD4+ T lymphocytes isolated from HIV-1-infected individuals receiving suppressive combined antiretroviral therapy. J Virol 90:9712-9724. https://doi.org/10.1128/JVI.00852-16.
19. Gill AL, Green SA, Abdullah S, Le Saout C, Pittaluga S, Chen H, Turnier R, Lifson J, Godin S, Qin J, Sneller MC, Cuillerot JM, Sabzevari H, Lane HC, Catalfamo M. 2016. Programed death-1/programed death-ligand 1 expression in lymph nodes of HIV infected patients: results of a pilot safety study in rhesus macaques using anti-programed deathligand 1 (Avelumab). AIDS 30:2487-2493. https://doi.org/10.1097/QAD.0000000000001217.
20. Lum JJ, Schnepple DJ, Nie Z, Sanchez-Dardon J, Mbisa GL, Mihowich J, Hawley N, Narayan S, Kim JE, Lynch DH, Badley AD. 2004. Differential effects of interleukin-7 and interleukin-15 on NK cell anti-human immunodeficiency virus activity. J Virol 78:6033-6042. https://doi.org/10.1128/JVI.78.11.6033-6042.2004.
21. Seay K, Church C, Zheng JH, Deneroff K, Ochsenbauer C, Kappes JC, Liu B, Jeng EK, Wong HC, Goldstein H. 2015. In vivo activation of human NK cells by treatment with an interleukin-15 superagonist potently inhibits acute in vivo HIV-1 infection in humanized mice. J Virol 89:6264-6274. https://doi.org/10.1128/JVI.00563-15.
22. Correia C, Lee SH, Meng XW, Vincelette ND, Knorr KL, Ding H, Nowakowski GS, Dai H, Kaufmann SH. 2015. Emerging understanding of Bcl-2 biology: implications for neoplastic progression and treatment. Biochim Biophys Acta 1853:1658-1671. https://doi.org/10.1016/j.bbamcr.2015.03.012.
23. Cummins NW, Sainski AM, Dai H, Natesampillai S, Pang YP, Bren GD, de Araujo Correia MC, Sampath R, Rizza SA, O'Brien D, Yao JD, Kaufmann SH, Badley AD. 2016. Prime, shock, and kill: priming CD4 T cells from HIV patients with a BCL-2 antagonist before HIV reactivation reduces HIV reservoir size. J Virol 90:4032-4048. https://doi.org/10.1128/JVI.03179-15.
24. Luna-Vargas MP, Chipuk JE. 2016. The deadly landscape of pro-apoptotic BCL-2 proteins in the outer mitochondrial membrane. FEBS J 283: 2676-2689. https://doi.org/10.1111/febs.13624.
25. Doerflinger M, Glab JA, Puthalakath H. 2015. BH3-only proteins: a 20-year stock-take. FEBS J 282:1006-1016. https://doi.org/10.1111/febs.13190.
26. Brenchley JM, Hill BJ, Ambrozak DR, Price DA, Guenaga FJ, Casazza JP, Kuruppu J, Yazdani J, Migueles SA, Connors M, Roederer M, Douek DC, Koup RA. 2004. T-cell subsets that harbor human immunodeficiency virus (HIV) in vivo: implications for HIV pathogenesis. J Virol 78: 1160-1168. https://doi.org/10.1128/JVI.78.3.1160-1168.2004.
27. Nie Z, Phenix BN, Lum JJ, Alam A, Lynch DH, Beckett B, Krammer PH, Sekaly RP, Badley AD. 2002. HIV-1 protease processes procaspase 8 to cause mitochondrial release of cytochrome c, caspase cleavage and nuclear fragmentation. Cell Death Differ 9:1172-1184. https://doi.org/10.1038/sj.cdd.4401094.
28. Nie Z, Bren GD, Vlahakis SR, Schimnich AA, Brenchley JM, Trushin SA, Warren S, Schnepple DJ, Kovacs CM, Loutfy MR, Douek DC, Badley AD. 2007 Human immunodeficiency virus type 1 protease cleaves procaspase 8 in vivo. J Virol 81:6947-6956. https://doi.org/10.1128/JVI.02798-06.
29. Sainski AM, Natesampillai S, Cummins NW, Bren GD, Taylor J, Saenz DT, Poeschla EM, Badley AD. 2011. The HIV-1-specific protein Casp8p41 induces death of infected cells through Bax/Bak. J Virol 85:7965-7975. https://doi.org/10.1128/JVI.02515-10.
30. Sainski AM, Dai H, Natesampillai S, Pang YP, Bren GD, Cummins NW, Correia C, Meng XW, Tarara JE, Ramirez-Alvarado M, Katzmann DJ, Ochsenbauer C, Kappes JC, Kaufmann SH, Badley AD. 2014. Casp8p41 generated by HIV protease kills CD4 T cells through direct Bak activation. J Cell Biol 206:867-876. https://doi.org/10.1083/jcb.201405051.
31. Nie Z, Bren GD, Rizza SA, Badley AD. 2008. HIV protease cleavage of procaspase 8 is necessary for death of HIV-infected cells. Open Virol J 2:1-7. https://doi.org/10.2174/1874357900802010001.
32. Jordan A, Bisgrove D, Verdin E. 2003. HIV reproducibly establishes a latent infection after acute infection of T cells in vitro. EMBO J 22: 1868-1877. https://doi.org/10.1093/emboj/cdg188.
33. Roberts AW, Huang D. 26 November 2016. Targeting BCL2 with BH3 mimetics: basic science and clinical application of venetoclax in chronic lymphocytic leukemia and related B cell malignancies. Clin Pharmacol Ther https://doi.org/10.1002/cpt.553.
34. Kim HR, Hwang KA, Kim KC, Kang I. 2007. Down-regulation of IL-7Ralpha expression in human T cells via DNA methylation. J Immunol 178: 5473-5479. https://doi.org/10.4049/jimmunol.178.9.5473.
35. Venkatachari NJ, Zerbato JM, Jain S, Mancini AE, Chattopadhyay A, Sluis-Cremer N, Bar-Joseph Z, Ayyavoo V. 2015. Temporal transcriptional response to latency reversing agents identifies specific factors regulating HIV-1 viral transcriptional switch. Retrovirology 12:85. https://doi.org/10.1186/s12977-015-0211-3.
36. Shan L, Deng K, Shroff NS, Durand CM, Rabi SA, Yang HC, Zhang H, Margolick JB, Blankson JN, Siliciano RF. 2012. Stimulation of HIV-1-specific cytolytic T lymphocytes facilitates elimination of latent viral reservoir after virus reactivation. Immunity 36:491-501. https://doi.org/10.1016/j.immuni.2012.01.014.
37. Davids MS, Roberts AW, Seymour JF, Pagel JM, Kahl BS, Wierda WG, Puvvada S, Kipps TJ, Anderson MA, Salem AH, Dunbar M, Zhu M, Peale F, Ross JA, Gressick L, Desai M, Kim SY, Verdugo M, Humerickhouse RA, Gordon GB, Gerecitano JF. 2017. Phase I first-in-human study of venetoclax in patients with relapsed or refractory non-Hodgkin lymphoma. J Clin Oncol 35:826-833. https://doi.org/10.1200/JCO.2016.70.4320.
38. Roberts AW, Davids MS, Pagel JM, Kahl BS, Puvvada SD, Gerecitano JF, Kipps TJ, Anderson MA, Brown JR, Gressick L, Wong S, Dunbar M, Zhu M, Desai MB, Cerri E, Heitner Enschede S, Humerickhouse RA, Wierda WG, Seymour JF. 2016. Targeting BCL2 with venetoclax in relapsed chronic lymphocytic leukemia. N Engl J Med 374:311-322. https://doi.org/10.1056/NEJMoa1513257.
39. Cummins NW, Sainski AM, Natesampillai S, Bren GD, Badley AD. 2014. Choice of antiretroviral therapy differentially impacts survival of HIVinfected CD4 T cells. Mol Cell Ther 2:1. https://doi.org/10.1186/2052-8426-2-1.
40. McDonnell TJ, Deane N, Platt FM, Nunez G, Jaeger U, McKearn JP, Korsmeyer SJ. 1989. bcl-2-immunoglobulin transgenic mice demonstrate extended B cell survival and follicular lymphoproliferation. Cell 57:79-88. https://doi.org/10.1016/0092-8674(89)90174-8.
41. Pegoraro L, Palumbo A, Erikson J, Falda M, Giovanazzo B, Emanuel BS, Rovera G, Nowell PC, Croce CM. 1984. A 14;18 and an 8;14 chromosome translocation in a cell line derived from an acute B-cell leukemia. Proc Natl Acad Sci U S A 81:7166-7170. https://doi.org/10.1073/pnas.81.22.7166.
42. Nuñez G, Hockenbery D, McDonnell TJ, Sorensen CM, Korsmeyer SJ. 1991 Bcl-2 maintains B cell memory. Nature 353:71-73. https://doi.org/10.1038/353071a0.
43. Yang HC, Xing S, Shan L, O'Connell K, Dinoso J, Shen A, Zhou Y, Shrum CK, Han Y, Liu JO, Zhang H, Margolick JB, Siliciano RF. 2009. Smallmolecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation. J Clin Invest 119:3473-3486. https://doi.org/10.1172/JCI39199.
44. Algeciras-Schimnich A, Belzacq-Casagrande AS, Bren GD, Nie Z, Taylor JA, Rizza SA, Brenner C, Badley AD. 2007. Analysis of HIV protease killing through caspase 8 reveals a novel interaction between caspase 8 and mitochondria. Open Virol J 1:39-46.
45. Cummins NW, Jiang W, McGinty J, Bren GD, Bosch RJ, Landay A, Deeks SG, Martin JN, Douek D, Lederman MM, Brenchley J, Badley AD. 2010. Intracellular Casp8p41 content is inversely associated with CD4 T cell count. J Infect Dis 202:386-391. https://doi.org/10.1086/653705.
46. Cummins NW, Neuhaus J, Sainski AM, Strausbauch MA, Wettstein PJ, Lewin SR, Plana M, Rizza SA, Temesgen Z, Touloumi G, Freiberg M, Neaton J, Badley AD. 2014. Short communication: CD4 T cell declines occurring during suppressive antiretroviral therapy reflect continued production of Casp8p41. AIDS Res Hum Retroviruses 30:476-479. https://doi.org/10.1089/aid.2013.0243.
47. Zauli G, Gibellini D, Caputo A, Bassini A, Negrini M, Monne M, Mazzoni M, Capitani S. 1995. The human immunodeficiency virus type-1 Tat protein upregulates Bcl-2 gene expression in Jurkat T-cell lines and primary peripheral blood mononuclear cells. Blood 86:3823-3834.
48. Wang Z, Morris GF, Reed JC, Kelly GD, Morris CB. 1999. Activation of Bcl-2 promoter-directed gene expression by the human immunodeficiency virus type-1 Tat protein. Virology 257:502-510. https://doi.org/10.1006/viro.1999.9688.
49. Zhang M, Li X, Pang X, Ding L, Wood O, Clouse KA, Hewlett I, Dayton AI. 2002 Bcl-2 upregulation by HIV-1 Tat during infection of primary human macrophages in culture. J Biomed Sci 9:133-139. https://doi.org/10.1007/BF02256024.
50. Zheng L, Yang Y, Guocai L, Pauza CD, Salvato MS. 2007. HIV Tat protein increases Bcl-2 expression in monocytes which inhibits monocyte apoptosis induced by tumor necrosis factor-alpha-related apoptosis-induced ligand. Intervirology 50:224-228. https://doi.org/10.1159/000100565.
51. Conti L, Rainaldi G, Matarrese P, Varano B, Rivabene R, Columba S, Sato A, Belardelli F, Malorni W, Gessani S. 1998. The HIV-1 Vpr protein acts as a negative regulator of apoptosis in a human lymphoblastoid T cell line: possible implications for the pathogenesis of AIDS. J Exp Med 187: 403-413. https://doi.org/10.1084/jem.187.3.403.
52. Chaudhary PM, Eby MT, Jasmin A, Kumar A, Liu L, Hood L. 2000. Activation of the NF-kappaB pathway by caspase 8 and its homologs. Oncogene 19:4451-4460. https://doi.org/10.1038/sj.onc.1203812.
53. Kretzschmar M, Meisterernst M, Scheidereit C, Li G, Roeder RG. 1992. Transcriptional regulation of the HIV-1 promoter by NF-kappa B in vitro. Genes Dev 6:761-774. https://doi.org/10.1101/gad.6.5.761.
54. Strack PR, Frey MW, Rizzo CJ, Cordova B, George HJ, Meade R, Ho SP, Corman J, Tritch R, Korant BD. 1996. Apoptosis mediated by HIV protease is preceded by cleavage of Bcl-2. Proc Natl Acad Sci U S A 93:9571-9576. https://doi.org/10.1073/pnas.93.18.9571.
55. Shoeman RL, Sachse C, Honer B, Mothes E, Kaufmann M, Traub P. 1993. Cleavage of human and mouse cytoskeletal and sarcomeric proteins by human immunodeficiency virus type 1 protease. Actin, desmin, myosin, and tropomyosin. Am J Pathol 142:221-230.
56. Shoeman RL, Huttermann C, Hartig R, Traub P. 2001. Amino-terminal polypeptides of vimentin are responsible for the changes in nuclear architecture associated with human immunodeficiency virus type 1 protease activity in tissue culture cells. Mol Biol Cell 12:143-154. https://doi.org/10.1091/mbc.12.1.143.
57. Rumlová M, Krížová I, Keprová A, Hadravová R, Doležal M, Strohalmová K, Pichová I, Hájek M, Ruml T. 2014. HIV-1 protease-induced apoptosis. Retrovirology 11:37. https://doi.org/10.1186/1742-4690-11-37.
58. Bren GD, Whitman J, Cummins N, Shepard B, Rizza SA, Trushin SA, Badley AD. 2008. Infected cell killing by HIV-1 protease promotes NF-kappaB dependent HIV-1 replication. PLoS One 3:e2112. https://doi.org/10.1371/journal.pone.0002112.
59. Maldarelli F, Wu X, Su L, Simonetti FR, Shao W, Hill S, Spindler J, Ferris AL, Mellors JW, Kearney MF, Coffin JM, Hughes SH. 2014. HIV latency. Specific HIV integration sites are linked to clonal expansion and persistence of infected cells. Science 345:179-183.
60. Antoni BA, Sabbatini P, Rabson AB, White E. 1995. Inhibition of apoptosis in human immunodeficiency virus-infected cells enhances virus production and facilitates persistent infection. J Virol 69:2384-2392.
61. Sandstrom PA, Pardi D, Goldsmith CS, Chengying D, Diamond AM, Folks TM. 1996. bc1-2 expression facilitates human immunodeficiency virus type-1 mediated cytopathic effects during acute spreading infections. J Virol 70:4617-4622.
62. Marshall WL, Datta R, Hanify K, Teng E, Finberg RW. 1999. U937 cells overexpressing bcl-xl are resistant to human immunodeficiency virus-1-induced apoptosis and human immunodeficiency virus-1 replication. Virology 256:1-7. https://doi.org/10.1006/viro.1999.9599.
63. Adachi Y, Oyaizu N, Than S, McCloskey TW, Pahwa S. 1996. IL-2 rescues in vitro lymphocyte apoptosis in patients with HIV infection: correlation with its ability to block culture-induced down-modulation of Bcl-2. J Immunol 157:4184-4193.
64. Chinnaiyan AM, Woffendin C, Dixit VM, Nabel GJ. 1997. The inhibition of pro-apoptotic ICE-like proteases enhances HIV replication. Nat Med 3:333-337. https://doi.org/10.1038/nm0397-333.
65. Badley AD, Sainski A, Wightman F, Lewin SR. 2013. Altering cell death pathways as an approach to cure HIV infection. Cell Death Dis 4:e718. https://doi.org/10.1038/cddis.2013.248.
66. Dietz AB, Bulur PA, Emery RL, Winters JL, Epps DE, Zubair AC, Vuk-Pavlovic S. 2006. A novel source of viable peripheral blood mononuclear cells from leukoreduction system chambers. Transfusion 46:2083-2089. https://doi.org/10.1111/j.1537-2995.2006.01033.x.
67. Single A, Beetham H, Telford BJ, Guilford P, Chen A. 2015. A comparison of real-time and endpoint cell viability assays for improved synthetic lethal drug validation. J Biomol Screen 20:1286-1293. https://doi.org/10.1177/1087057115605765.
68. Cummins NW, Klicpera A, Sainski AM, Bren GD, Khosla S, Westendorf JJ, Badley AD. 2011. Human immunodeficiency virus envelope protein Gp120 induces proliferation but not apoptosis in osteoblasts at physiologic concentrations. PLoS One 6:e24876. https://doi.org/10.1371/journal.pone.0024876.