PD-1 AND TIM-3 BLOCKING CANNOT ENHANCE APOPTOSIS OF CHRONIC LYMPHOCYTIC LEUKEMIA CELLS INDUCED BY PERIPHERAL BLOOD CD8+ T CELLS
DOI:
https://doi.org/10.32471/exp-oncology.2312-8852.vol-44-no-4.18975Keywords:
anti-PD-1, anti-TIM-3, apoptosis, CD8+ T cells, CLLAbstract
Aim: Given the invaluable success of immune checkpoint inhibitors for tumor immunotherapy, in this study, the effect of programmed cell death 1 (PD-1) and T cell immunoglobulin-3 (TIM-3) blocking was investigated to induce apoptosis of leukemic cells by exhausted CD8+ T cells in patients with chronic lymphocytic leukemia (CLL). Materials and Methods: Peripheral blood CD8+ T cells were positively isolated from 16 CLL patients using magnetic beads separation method. Isolated CD8+ T cells were treated with either blocking anti-PD-1, anti-TIM-3 and isotype-matched control antibodies and then co-cultured with CLL leukemic cells as target. The percentage of apoptotic leukemic cells and the expression of apoptosis-related genes were evaluated by flow cytometry and real-time polymerase chain reaction methods, respectively. Interferon gamma and tumor necrosis factor alpha concentration was also measured by ELISA. Results: Flow cytometric analysis of apoptotic leukemic cells indicated that the blockade of PD-1 and TIM-3 did not significantly enhance the apoptosis of CLL cells by CD8+T cells, which then were confirmed by analysis of BAX, BCL2 and CASP3 gene expression, which was similar in blocked and control groups. No significant difference was found between blocked and control groups in terms of interferon gamma and tumor necrosis factor alpha production by CD8+ T cells. Conclusion: We concluded that the blockade of PD-1 and TIM-3 is not an effective strategy to restore the function of CD8+ T cells in CLL patients at the early clinical stages of the disease. Further in vitro and in vivo studies are needed to more address the application of immune checkpoint blockade in CLL patients.
References
Bosch F, Dalla-Favera R. Chronic lymphocytic leukaemia: from genetics to treatment. Nat Rev Clin Oncol 2019; 16: 684–701. doi: https://doi.org/10.1038/s41571-019-0239-8
Kipps TJ, Stevenson FK, Wu CJ, et al. Chronic lymphocytic leukaemia. Nat Rev Dis Primers 2017; 3: 17008. doi: https://doi.org/10.1038/nrdp.2017.8
Bagacean C, Tomuleasa C, Tempescul A, et al. Apoptotic resistance in chronic lymphocytic leukemia and therapeutic perspectives. Crit Rev Clin Lab Sci 2019; 56: 321–32. doi: https://doi.org/10.1080/10408363.2019.1600468
Hude I, Sasse S, Engert A, et al. The emerging role of immune checkpoint inhibition in malignant lymphoma. Haematologica 2017; 102: 30–42. doi: 10.3324/haematol.2016.150656
Chen P-H, Ho C-L, Lin C, et al. Treatment outcomes of novel targeted agents in relapse/refractory chronic lymphocytic leukemia: A systematic review and network meta-analysis. J Clin Med 2019; 8: 737. doi: 10.3390/jcm8050737
McClanahan F, Riches JC, Miller S, et al. Mechanisms of PD-L1/PD-1–mediated CD8 T-cell dysfunction in the context of aging-related immune defects in the Eµ-TCL1 CLL mouse model. Blood 2015; 126: 212–21. doi: https://doi.org/10.1182/blood-2015-02-626754
Lewinsky H, Barak AF, Huber V, et al. CD84 regulates PD-1/PD-L1 expression and function in chronic lymphocytic leukemia. J Clin Invest 2018; 128: 5465–78. doi: https://doi.org/10.1172/JCI96610
McLane LM, Abdel-Hakeem MS, Wherry EJ. CD8 T cell exhaustion during chronic viral infection and cancer. Annu Rev Immunol 2019; 37: 457–95. doi: https://doi.org/10.1146/annurev-immunol-041015-055318
Dyck L, Mills KH. Immune checkpoints and their inhibition in cancer and infectious diseases. Eur J Immunol 2017; 47: 765–79. doi: https://doi.org/10.1002/eji.201646875
Schietinger A, Greenberg PD. Tolerance and exhaustion: defining mechanisms of T cell dysfunction. Trends Immunol 2014; 35: 51–60. doi: https://doi.org/10.1016/j.it.2013.10.001
Taghiloo S, Allahmoradi E, Tehrani M, et al. Frequency and functional characterization of exhausted CD8+ T-cells in chronic lymphocytic leukemia. Eur J Haematol 2017; 98: 622–31. doi: https://doi.org/10.1111/ejh.12880
Shapiro M, Herishanu Y, Katz BZ, et al. Lymphocyte activation gene 3: a novel therapeutic target in chronic lymphocytic leukemia. Haematologica 2017; 102: 874–82. doi: 10.3324/haematol.2016.148965
Wolchok JD, Hodi FS, Weber JS, et al. Development of ipilimumab: a novel immunotherapeutic approach for the treatment of advanced melanoma. Ann N Y Acad Sci 2013; 1291: 1–13. doi: https://doi.org/10.1111/nyas.12180
Robert C, Long GV, Brady B, et al. Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med 2015; 372: 320–30. doi: https://doi.org/10.1056/NEJMoa1412082
Brahmer J, Reckamp KL, Baas P, et al. Nivolumab versus docetaxel in advanced squamous-cell non–small-cell lung cancer. N Engl J Med 2015; 373: 123–35. doi: https://doi.org/10.1056/NEJMoa1504627
Motzer RJ, Escudier B, McDermott DF, et al. Nivolumab versus everolimus in advanced renal-cell carcinoma. N Engl J Med 2015; 373: 1803–13. doi: https://doi.org/10.1056/NEJMoa1510665
Ansell SM, Lesokhin AM, Borrello I, et al. PD-1 blockade with nivolumab in relapsed or refractory Hodgkin’s lymphoma. N Engl J Med 2015; 372: 311–9. doi: https://doi.org/10.1056/NEJMoa1411087
Ferris RL, Blumenschein Jr G, Fayette J, et al. Nivolumab for recurrent squamous-cell carcinoma of the head and neck. N Engl J Med 2016; 375: 1856–67. doi: https://doi.org/10.1056/NEJMoa1602252
Sharma P, Retz M, Siefker-Radtke A, et al. Nivolumab in metastatic urothelial carcinoma after platinum therapy (CheckMate 275): a multicentre, single-arm, phase 2 trial. Lancet Oncol 2017; 18: 312–22. doi: https://doi.org/10.1016/S1470-2045(17)30065-7
Overman MJ, McDermott R, Leach JL, et al. Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study. Lancet Oncol 2017; 18: 1182–91. doi: https://doi.org/10.1016/S1470-2045(17)30422-9
El-Khoueiry AB, Sangro B, Yau T, et al. Nivolumab in patients with advanced hepatocellular carcinoma (CheckMate 040): an open-label, non-comparative, phase 1/2 dose escalation and expansion trial. Lancet 2017; 389: 2492–502. doi: https://doi.org/10.1016/S0140-6736(17)31046-2
Chen R, Zinzani PL, Fanale MA, et al. Phase II study of the efficacy and safety of pembrolizumab for relapsed/refractory classic Hodgkin lymphoma. J Clin Oncol 2017; 35: 2125–32. doi: https://doi.org/10.1200/JCO.2016.72.1316
Herbst RS, Baas P, Kim D-W, et al. Pembrolizumab versus docetaxel for previously treated, PD-L1-positive, advanced non-small-cell lung cancer (KEYNOTE-010): a randomised controlled trial. Lancet 2016; 387: 1540–50. doi: https://doi.org/10.1016/S0140-6736(15)01281-7
Bellmunt J, De Wit R, Vaughn DJ, et al. Pembrolizumab as second-line therapy for advanced urothelial carcinoma. N Engl J Med 2017; 376: 1015–26. doi: https://doi.org/10.1056/NEJMoa1613683
Fuchs CS, Doi T, Jang RW, et al. Safety and efficacy of pembrolizumab monotherapy in patients with previously treated advanced gastric and gastroesophageal junction cancer: phase 2 clinical KEYNOTE-059 trial. JAMA Oncol 2018; 4: e180013. doi: https://doi.org/10.1001/jamaoncol.2018.0013
Powles T, Durán I, Van Der Heijden MS, et al. Atezolizumab versus chemotherapy in patients with platinum-treated locally advanced or metastatic urothelial carcinoma (IMvigor211): a multicentre, open-label, phase 3 randomised controlled trial. Lancet 2018; 391: 748–57. doi: https://doi.org/10.1016/S0140-6736(17)33297-X
Patel MR, Ellerton J, Infante JR, et al. Avelumab in metastatic urothelial carcinoma after platinum failure (JAVELIN Solid Tumor): pooled results from two expansion cohorts of an open-label, phase 1 trial. Lancet Oncol 2018; 19: 51–64. doi: https://doi.org/10.1016/S1470-2045(17)30900-2
Antonia SJ, Villegas A, Daniel D, et al. Durvalumab after chemoradiotherapy in stage III non–small-cell lung cancer. N Engl J Med 2017; 377: 1919–29. doi: https://doi.org/10.1056/NEJMoa1709937
Rittmeyer A, Barlesi F, Waterkamp D, et al. Atezolizumab versus docetaxel in patients with previously treated non-small-cell lung cancer (OAK): a phase 3, open-label, multicentre randomised controlled trial. Lancet 2017; 389: 255–65. doi: https://doi.org/10.1016/S0140-6736(16)32517-X
Kaufman HL, Russell J, Hamid O, et al. Avelumab in patients with chemotherapy-refractory metastatic Merkel cell carcinoma: a multicentre, single-group, open-label, phase 2 trial. Lancet Oncol 2016; 17: 1374–85. doi: https://doi.org/10.1016/S1470-2045(16)30364-3
Du W, Yang M, Turner A, et al. TIM-3 as a target for cancer immunotherapy and mechanisms of action. Int J Mol Sci 2017; 18: 645. doi: 10.3390/ijms18030645
Allahmoradi E, Taghiloo S, Omrani-Nava V, et al. Anti-inflammatory effects of the Portulaca oleracea hydroalcholic extract on human peripheral blood mononuclear cells. Med J Islam Repub Iran 2018; 32: 80. doi: https://doi.org/10.14196/mjiri.32.80
Taghiloo S, Allahmoradi E, Ebadi R, et al. Upregulation of Galectin-9 and PD-L1 immune checkpoints molecules in patients with chronic lymphocytic leukemia. Asian Pac J Cancer Prev 2017; 18: 2269–74. doi: 10.22034/APJCP.2017.18.8.2269.
Rezazadeh H, Astaneh M, Tehrani M, et al. Blockade of PD-1 and TIM-3 immune checkpoints fails to restore the function of exhausted CD8+ T cells in early clinical stages of chronic lymphocytic leukemia. Immunol Res 2020; 68: 269–79. doi: https://doi.org/10.1007/s12026-020-09146-4
Jones RB, Ndhlovu LC, Barbour JD, et al. Tim-3 expression defines a novel population of dysfunctional T cells with highly elevated frequencies in progressive HIV-1 infection. J Exp Med 2008; 205: 2763–79. doi: https://doi.org/10.1084/jem.20081398
Lu X, Yang L, Yao D, et al. Tumor antigen-specific CD8+ T cells are negatively regulated by PD-1 and Tim-3 in human gastric cancer. Cell Immunol 2017; 313: 43–51. doi: https://doi.org/10.1016/j.cellimm.2017.01.001
Fourcade J, Sun Z, Benallaoua M, et al. Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen–specific CD8+ T cell dysfunction in melanoma patients. J Exp Med 2010; 207: 2175–86. doi: https://doi.org/10.1084/jem.20100637
Barber DL, Wherry EJ, Masopust D, et al. Restoring function in exhausted CD8 T cells during chronic viral infection. Nature 2006; 439: 682–7. doi: https://doi.org/10.1038/nature04444
Trautmann L, Janbazian L, Chomont N, et al. Upregulation of PD-1 expression on HIV-specific CD8+ T cells leads to reversible immune dysfunction. Nat Med 2006; 12: 1198–202. doi: https://doi.org/10.1038/nm1482
Zhang L, Gajewski TF, Kline J. PD-1/PD-L1 interactions inhibit antitumor immune responses in a murine acute myeloid leukemia model. Blood 2009; 114: 1545–52. doi: https://doi.org/10.1182/blood-2009-03-206672
Yamamoto R, Nishikori M, Kitawaki T, et al. PD-1–PD-1 ligand interaction contributes to immunosuppressive microenvironment of Hodgkin lymphoma. Blood 2008; 111: 3220–4. doi: https://doi.org/10.1182/blood-2007-05-085159
Mumprecht S, Schürch C, Schwaller J, et al. Programmed death 1 signaling on chronic myeloid leukemia–specific T cells results in T-cell exhaustion and disease progression. Blood 2009; 114: 1528–36. doi: https://doi.org/10.1182/blood-2008-09-179697
Takamura S, Tsuji-Kawahara S, Yagita H, et al. Premature terminal exhaustion of Friend virus-specific effector CD8+ T cells by rapid induction of multiple inhibitory receptors. J Immunol 2010; 184: 4696–707. doi: 10.4049/jimmunol.0903478
Sakuishi K, Apetoh L, Sullivan JM, et al. Targeting Tim-3 and PD-1 pathways to reverse T cell exhaustion and restore anti-tumor immunity. J Exp Med 2010; 207: 2187–94. doi: https://doi.org/10.1084/jem.20100643
Duraiswamy J, Kaluza KM, Freeman GJ, et al. Dual blockade of PD-1 and CTLA-4 combined with tumor vaccine effectively restores T-cell rejection function in tumors. Cancer Res 2013; 73: 3591–603. doi: https://doi.org/10.1158/0008-5472.CAN-12-4100
Riches JC, Davies JK, McClanahan F, et al. T cells from CLL patients exhibit features of T-cell exhaustion but retain capacity for cytokine production. Blood 2013; 121: 1612–21. doi: https://doi.org/10.1182/blood-2012-09-457531
Behdad A, Griffin B, Chen YH, et al. PD-1 is highly expressed by neoplastic B-cells in Richter transformation. Br J Haematol 2019; 185: 370–3. doi: https://doi.org/10.1111/bjh.15514
Younes A, Brody J, Carpio C, et al. Safety and activity of ibrutinib in combination with nivolumab in patients with relapsed non-Hodgkin lymphoma or chronic lymphocytic leukaemia: a phase 1/2a study. Lancet Haematol 2019; 6: e67–78. doi: https://doi.org/10.1016/S2352-3026(18)30217-5
Ding W, Le-Rademacher J, Call TG, et al. PD-1 blockade with pembrolizumab in relapsed CLL including Richter’s transformation: an updated report from a phase 2 trial (MC1485). Blood 2016; 128: 4392. doi: https://doi.org/10.1182/blood.V128.22.4392.4392
Jain N, Ferrajoli A, Basu S, et al. A phase II trial of Nivolumab combined with Ibrutinib for patients with Richter transformation. Blood 2018; 132: 296. doi: https://doi.org/10.1182/blood-2018-99-120355
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