Involvement of MM cell‑derived exosomes in T lymphocytes immune responses

Shao,Qing; Deng,Ling; Liu,Hui; Liu,Zhaoyun; Chen,Jin; Jiang,Fengjuan; Yan,Siyang; Fu,Rong

doi:10.3892/ol.2020.11892

Involvement of MM cell‑derived exosomes in T lymphocytes immune responses

Authors:
- Qing Shao
- Ling Deng
- Hui Liu
- Zhaoyun Liu
- Jin Chen
- Fengjuan Jiang
- Siyang Yan
- Rong Fu
View Affiliations

Affiliations: Department of Hematology, Tianjin Medical University General Hospital, Heping, Tianjin 300052, P.R. China
Published online on: July 17, 2020 https://doi.org/10.3892/ol.2020.11892
Article Number: 31
Copyright: © Shao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Exosomes were reported to mediate cell communication in the tumor microenvironment; however, the effects of multiple myeloma (MM)‑derived exosomes on the quantity and function of T cells remain unknown. Exosomes were extracted from MM cell lines (OPM2 and U266B1) by ultracentrifugation using a Total Exosome Isolation kit. Exosomes were co‑cultured with CD4+ T, CD8+ T and regulatory T (Treg) cells that were isolated from healthy donors (HDs) and patients with MM using magnetic beads. Flow cytometry was used to detect T cells apoptosis and expression of perforin and granzyme B in CD8+ T cells. Cell viability was detected using Cell Counting kit‑8, and interleukin 10 (IL‑10) and transforming growth factor β (TGF‑β) in cell supernatants were detected by ELISA. The apoptosis of HD‑CD4+ T was higher in the OPM2 group, and viability in the U266B1 group was decreased. The apoptosis of HD‑CD8+ T decreased in the OPM2 and U266B1 groups, and cell viability increased in the OPM2 and the U266B1 groups. Perforin of HD‑CD8+ T in the U266B1 group was lower while perforin of MM‑CD8+ T in OPM2 and U266B1 groups was markedly decreased. The apoptosis of HD‑Treg was lower in the U266B1 group, but apoptosis of MM‑Treg was higher in the U266B1 group. The viability of HD‑Treg in U266B1 group increased but the viability of MM‑Treg in OPM2 and U266B1 groups decreased. TGF‑β from MM‑Treg decreased in the OPM2 and U266B1 groups when compared with the control group (P<0.05). MM‑derived exosomes promote apoptosis and inhibit proliferation of HD‑CD4+ T, inhibit apoptosis and promote proliferation, but inhibit perforin of HD‑CD8+ T, inhibit apoptosis and promote proliferation HD‑Treg, and inhibit perforin of MM‑CD8+ T and TGF‑β secretion of MM‑Treg.

View Figures

View References

1	Rajkumar SV, Dimopoulos MA, Palumbo A, Blade J, Merlini G, Mateos MV, Kumar S, Hillengass J, Kastritis E, Richardson P, et al: International Myeloma Working Group updated criteria for the diagnosis of multiple myeloma. Lancet Oncol. 15:e538–e548. 2014. View Article : Google Scholar : PubMed/NCBI
2	Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J, Rosso S, Coebergh JW, Comber H, Forman D and Bray F: Cancer incidence and mortality patterns in Europe: Estimates for 40 countries in 2012. Eur J Cancer. 49:1374–1403. 2013. View Article : Google Scholar : PubMed/NCBI
3	Kumar SK, Dispenzieri A, Lacy MQ, Gertz MA, Buadi FK, Pandey S, Kapoor P, Dingli D, Hayman SR, Leung N, et al: Continued improvement in survival in multiple myeloma: Changes in early mortality and outcomes in older patients. Leukemia. 28:1122–1128. 2014. View Article : Google Scholar : PubMed/NCBI
4	Mikkilineni L and Kochenderfer JN: Chimeric antigen receptor T-cell therapies for multiple myeloma. Blood. 130:2594–2602. 2017. View Article : Google Scholar : PubMed/NCBI
5	Zhang X, Yuan X, Shi H, Wu L, Qian H and Xu W: Exosomes in cancer: Small particle, big player. J Hematol Oncol. 8:832015. View Article : Google Scholar : PubMed/NCBI
6	Yu S, Cao H, Shen B and Feng J: Tumor-derived exosomes in cancer progression and treatment failure. Oncotarget. 6:37151–37168. 2015. View Article : Google Scholar : PubMed/NCBI
7	Milane L, Singh A, Mattheolabakis G, Suresh M and Amiji MM: Exosome mediated communication within the tumor microenvironment. J Control Release. 219:278–294. 2015. View Article : Google Scholar : PubMed/NCBI
8	Yu S, Liu C, Su K, Wang J, Liu Y, Zhang L, Li C, Cong Y, Kimberly R, Grizzle WE, et al: Tumor exosomes inhibit differentiation of bone marrow dendritic cells. J Immunol. 178:6867–6875. 2007. View Article : Google Scholar : PubMed/NCBI
9	Clayton A, Mitchell JP, Court J, Mason MD and Tabi Z: Human tumor-derived exosomes selectively impair lymphocyte responses to interleukin-2. Cancer Res. 67:7458–7466. 2007. View Article : Google Scholar : PubMed/NCBI
10	Szajnik M, Czystowska M, Szczepanski MJ, Mandapathil M and Whiteside TL: Tumor-derived microvesicles induce, expand and up-regulate biological activities of human regulatory T cells (Treg). PLoS One. 5:e114692010. View Article : Google Scholar : PubMed/NCBI
11	Chalmin F, Ladoire S, Mignot G, Vincent J, Bruchard M, Remy-Martin JP, Boireau W, Rouleau A, Simon B, Lanneau D, et al: Membrane-associated Hsp72 from tumor-derived exosomes mediates STAT3-dependent immune suppressive function of mouse and human myeloid-derived suppressor cells. J Clin Invest. 120:457–471. 2010.PubMed/NCBI
12	Xiang X, Poliakov A, Liu C, Liu Y, Deng ZB, Wang J, Cheng Z, Shah SV, Wang GJ, Zhang L, et al: Induction of myeloid-derived suppressor cells by tumor exosomes. Int J Cancer. 124:2621–2633. 2009. View Article : Google Scholar : PubMed/NCBI
13	Muller L, Mitsuhashi M, Simms P, Gooding WE and Whiteside TL: Tumor-derived exosomes regulate expression of immune function related genes in human T cell subsets. Sci Rep. 6:202542016. View Article : Google Scholar : PubMed/NCBI
14	Wang J, De Veirman K, Faict S, Frassanito MA, Ribatti D, Vacca A and Menu E: Multiple myeloma exosomes establish a favourable bone marrow microenvironment with enhanced angiogenesis and immunosuppression. J Pathol. 239:162–173. 2016. View Article : Google Scholar : PubMed/NCBI
15	Raimondi L, De Luca A, Amodio N, Manno M, Raccosta S, Taverna S, Bellavia D, Naselli F, Fontana S, Schillaci O, et al: Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation. Oncotarget. 6:13772–13789. 2015. View Article : Google Scholar : PubMed/NCBI
16	Vignali DAA, Collison LW and Workman CJ: How regulatory T cells work. Nat Rev Immunol. 8:523–532. 2008. View Article : Google Scholar : PubMed/NCBI
17	Andersen MH, Schrama D, Thor Straten P and Becker JC: Cytotoxic T cells. J Invest Dermatol. 126:32–41. 2006. View Article : Google Scholar : PubMed/NCBI
18	Théry C, Amigorena S, Raposo G and Clayton A: Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol Chapter. 3:Unit 3.22. 2006.
19	Wang J, Hendrix A, Hernot S, Lemaire M, De Bruyne E, Van Valckenborgh E, Lahoutte T, De Wever O, Vanderkerken K and Menu E: Bone marrow stromal cell-derived exosomes as communicators in drug resistance in multiple myeloma cells. Blood. 124:555–566. 2014. View Article : Google Scholar : PubMed/NCBI
20	Jung MK and Mun JY: Sample preparation and imaging of exosomes by transmission electron microscopy. J Vis Exp. 131:564822018.
21	Quispe EÁ, Li XM and Yi H: Comparison and relationship of thyroid hormones, IL-6, IL-10 and albumin as mortality predictors in Case-mix critically ill patients. Cytokine. 81:94–100. 2016. View Article : Google Scholar : PubMed/NCBI
22	Lobb RJ, Becker M, Wen SW, Wong CS, Wiegmans AP, Leimgruber A and Möller A: Optimized exosome isolation protocol for cell culture supernatant and human plasma. J Extracell Vesicles. 4:270312015. View Article : Google Scholar : PubMed/NCBI
23	Vojtech L, Woo S, Hughes S, Levy C, Ballweber L, Sauteraud RP, Strobl J, Westerberg K, Gottardo R, Tewari M and Hladik F: Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions. Nucleic Acids Res. 42:7290–7304. 2014. View Article : Google Scholar : PubMed/NCBI
24	Boyiadzis M and Whiteside TL: The emerging roles of tumor-derived exosomes in hematological malignancies. Leukemia. 31:1259–1268. 2017. View Article : Google Scholar : PubMed/NCBI
25	Whiteside TL: Immune responses to malignancies. J Allergy Clin Immunol. 125 (Suppl):S272–S283. 2010. View Article : Google Scholar : PubMed/NCBI
26	Ludwig S, Floros T, Theodoraki MN, Hong CS, Jackson EK, Lang S and Whiteside TL: Suppression of lymphocyte functions by plasma exosomes correlates with disease activity in patients with head and neck cancer. Clin Cancer Res. 23:4843–4854. 2017. View Article : Google Scholar : PubMed/NCBI
27	Kim JW, Wieckowski E, Taylor DD, Reichert TE, Watkins S and Whiteside TL: Fas ligand-positive membranous vesicles isolated from sera of patients with oral cancer induce apoptosis of activated T lymphocytes. Clin Cancer Res. 11:1010–1020. 2005.PubMed/NCBI
28	Wieckowski EU, Visus C, Szajnik M, Szczepanski MJ, Storkus WJ and Whiteside TL: Tumor-derived microvesicles promote regulatory T cell expansion and induce apoptosis in tumor-reactive activated CD8+T lymphocytes. J Immunol. 183:3720–3730. 2009. View Article : Google Scholar : PubMed/NCBI
29	Kim JW, Tsukishiro T, Johnson JT and Whiteside TL: Expression of pro- and antiapoptotic proteins in circulating CD8+ T cells of patients with squamous cell carcinoma of the head and neck. Clin Cancer Res. 10:5101–5110. 2004. View Article : Google Scholar : PubMed/NCBI
30	Pardoll DM: The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer. 12:252–264. 2012. View Article : Google Scholar : PubMed/NCBI
31	Tran TH, Mattheolabakis G, Aldawsari H and Amiji M: Exosomes as nanocarriers for immunotherapy of cancer and inflammatory diseases. Clin Immunol. 160:46–58. 2015. View Article : Google Scholar : PubMed/NCBI
32	Ye SB, Li ZL, Luo DH, Huang BJ, Chen YS, Zhang XS, Cui J, Zeng YX and Li J: Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma. Oncotarget. 5:5439–5452. 2014. View Article : Google Scholar : PubMed/NCBI
33	Liu Y, Gu Y and Cao X: The exosomes in tumor immunity. Oncoimmunology. 4:e10274722015. View Article : Google Scholar : PubMed/NCBI
34	Mills KH and Cawley JC: Abnormal monoclonal antibody defined helper/suppressor T-cell subpopulations in multiple myeloma: Relationship to treatment and clinical stage. Br J Haematol. 53:271–275. 1983. View Article : Google Scholar : PubMed/NCBI
35	Prabhala RH, Neri P, Bae JE, Tassone P, Shammas MA, Allam CK, Daley JF, Chauhan D, Blanchard E, Thatte HS, et al: Dysfunctional T regulatory cells in multiple myeloma. Blood. 107:301–304. 2006. View Article : Google Scholar : PubMed/NCBI
36	Bryant C, Suen H, Brown R, Yang S, Favaloro J, Aklilu E, Gibson J, Ho PJ, Iland H, Fromm P, et al: Long-term survival in multiple myeloma is associated with a distinct immunological profile, which includes proliferative cytotoxic T-cell clones and a favourable Treg/Th17 balance. Blood Cancer J. 3:e1482013. View Article : Google Scholar : PubMed/NCBI
37	Beyer M, Kochanek M, Giese T, Endl E, Weihrauch MR, Knolle PA, Classen S and Schultze JL: In vivo peripheral expansion of naive CD4+CD25 high FoxP3+ regulatory T cells in patients with multiple myeloma. Blood. 107:3940–3949. 2006. View Article : Google Scholar : PubMed/NCBI
38	Giannopoulos K, Kaminska W, Hus I and Dmoszynska A: The frequency of T regulatory cells modulates the survival of multiple myeloma patients: Detailed characterisation of immune status in multiple myeloma. Br J Cancer. 106:546–552. 2012. View Article : Google Scholar : PubMed/NCBI
39	Feng P, Yan R, Dai X, Xie X, Wen H and Yang S: The alteration and clinical significance of Th1/Th2/Th17/Treg cells in patients with multiple myeloma. Inflammation. 38:705–709. 2015. View Article : Google Scholar : PubMed/NCBI
40	Andre F, Schartz NE, Chaput N, Flament C, Raposo G, Amigorena S, Angevin E and Zitvogel L: Tumor-derived exosomes: A new source of tumor rejection antigens. Vaccine. 20 (Suppl 4):A28–A31. 2002. View Article : Google Scholar : PubMed/NCBI
41	Natasha G, Gundogan B, Tan A, Farhatnia Y, Wu W, Rajadas J and Seifalian AM: Exosomes as immunotheranostic nanoparticles. Clin Ther. 36:820–829. 2014. View Article : Google Scholar : PubMed/NCBI
42	Xie Y, Bai O, Zhang H, Li W and Xiang J: Tumor necrosis factor gene-engineered J558 tumor cell-released exosomes stimulate tumor antigen P1A-specifc CD8⁺ CTL responses and antitumor immunity. Cancer Biother Radiopharm. 25:21–28. 2010. View Article : Google Scholar : PubMed/NCBI