Malp‑2, an agonist of tlr6, promotes the immune status without affecting the differentiation capacity of umbilical cord mesenchymal stem cells

Wu,Xiuli; Xu,Lian; Shen,Yangmei; Yu,Na; Zhang,Yan; Guo,Tao

doi:10.3892/etm.2017.5262

Malp‑2, an agonist of tlr6, promotes the immune status without affecting the differentiation capacity of umbilical cord mesenchymal stem cells

Authors:
- Xiuli Wu
- Lian Xu
- Yangmei Shen
- Na Yu
- Yan Zhang
- Tao Guo
View Affiliations

Affiliations: Department of Laboratory Medicine, West China Second Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Key Laboratory of Birth Defects and Related Diseases of Women and Children, Sichuan University, Ministry of Education, Beijing 100816, P.R. China
Published online on: October 3, 2017 https://doi.org/10.3892/etm.2017.5262
Pages: 5540-5546

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

Mesenchymal stem cells (MSCs) are increasingly used in cell‑based therapy due to their multiple differentiation capacity, low expression of co‑stimulatory factors and immunosuppressive effect. However, accumulating studies reported the recognition and rejection of engrafted MSCs, which eventually led to the fail of clinical trials. Toll‑like receptors (TLRs) are important in mediating the immune response. In the present study, macrophage‑activated lipopeptide‑2 (MALP‑2) was introduced to activate the TLR6 pathway in umbilical cord MSCs (UCMSCs). PBLs isolated from healthy volunteers were co‑cultured with UCMSCs to measure whether activation of TLR6 of UCMSCs could stimulate immune responses. Reverse transcription‑quantitative polymerase chain reaction and immunohistochemistry were performed to detect pro‑inflammatory molecules and differentiation status of UCMSCs, respectively. The results indicated that activation of TLR6 in UCMSCs increased the proliferation of peripheral blood leukocytes (PBLs) and enhanced the release of lactate dehydrogenase in damaged UCMSCs, which confirmed the role of TLR6 in promoting the immunogenicity of UCMSCs. Furthermore, quantitative polymerase chain reaction demonstrated that the expression of proinflammatory molecules (including IL‑1β, IL‑6, IL‑8, IL‑10, CCL1 and CCL4) was induced, whereas the expression of stem cell markers (Klf4 and Nanog) was inhibited. The differentiation results indicated that activation of TLR6 had no effect on the differentiation capacity of UCMSCs. All these findings suggest that stimulation of TLR6 pathway may increase the immunogenicity of UCMSCs in in vitro detections. In conclusion, the results of the current study indicated a new role of TLR6 in regulating the biological function of UCMSCs.

View Figures

View References

1	Le Blanc K, Frassoni F, Ball L, Locatelli F, Roelofs H, Lewis I, Lanino E, Sundberg B, Bernardo ME, Remberger M, et al: Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: A phase II study. Lancet. 371:1579–1586. 2008. View Article : Google Scholar : PubMed/NCBI
2	Vojtassak J, Danisovic L, Kubes M, Bakos D, Jarabek L, Ulicna M and Blasko M: Autologous biograft and mesenchymal stem cells in treatment of the diabetic foot. Neuro Endocrinol Lett. 27 Suppl 2:S134–S137. 2006.
3	Guiducci S, Porta F, Saccardi R, Guidi S, Ibba-Manneschi L, Manetti M, Mazzanti B, DalPozzo S, Milia AF, Bellando-Randone S, et al: Autologus mesenchymal stem cells foster revascularization of ischemic limbs in systemic sclerosis: A case report. Ann Intern Med. 153:650–654. 2010. View Article : Google Scholar : PubMed/NCBI
4	Bordignon C, Carlo-Stella C, Colombo MP, De Vincentiis A, Lanata L, Lemoli RM, Locatelli F, Olivieri A, Rondelli D, Zanon P and Tura S: Cell therapy: Achievements and perspectives. Haematologica. 84:1110–1149. 2011.
5	Phinney DG and Prockop DJ: Concise review: Mesenchymal stem/multipotent stromal cells: The state of transdifferentiation and modes of tissue repair-current views. Stem Cells. 25:2896–2902. 2007. View Article : Google Scholar : PubMed/NCBI
6	van Poll D, Parekkadan B, Cho CH, Berthiaume F, Nahmias Y, Tilles AW and Yarmuch ML: Mesenchymal stem cell-derived molecules directly modulate hepatocellular death and regeneration in vitro and in vivo. Hepatology. 47:1634–1643. 2008. View Article : Google Scholar : PubMed/NCBI
7	Shi M, Liu ZW and Wang FS: Immunomodulatory properties and therapeutic application of mesenchymal stem cells. Clin Exp Immunol. 164:1–8. 2011. View Article : Google Scholar : PubMed/NCBI
8	Han KH, Ro H, Hong JH, Lee EM, Cho B, Yeom HJ, Kim MG, Oh KH, Ahn C and Yang J: Immunosuppressive mechanisms of embryonic stem cells and mesenchymal stem cells in alloimmune response. Transpl Immunol. 25:7–15. 2011. View Article : Google Scholar : PubMed/NCBI
9	Bassi E, Aita CA and Câmara NO: Immune regulatory properties of multipotent mesenchymal stromal cells: Where do we stand? World J Stem Cells. 3:1–8. 2011. View Article : Google Scholar : PubMed/NCBI
10	Cui J, Wahl RL, Shen T, Fisher SJ, Recker E, Ginsburg D and Long MW: Bone marrow cell trafficking following intraveneous administration. Br J Haematol. 107:895–902. 1999. View Article : Google Scholar : PubMed/NCBI
11	Ankrum J and Karp JM: Mesenchymal stem cell therapy: Two steps forward, one step back. Trends Mol Med. 16:203–209. 2010. View Article : Google Scholar : PubMed/NCBI
12	Allison M: Genzyme back Osiris, despite Prochymal flop. Nat Biotechnol. 27:966–967. 2009. View Article : Google Scholar : PubMed/NCBI
13	Spaggiari GM, Capobianco A, Becchetti S, Mingari MC and Moretta L: Mesenchymal stem cell-natural killer cell interactions: Evidence that activated NK cells are capable of killing MSCs, whereas MSCs can inhibit IL-2-induced NK-cell proliferation. Blood. 107:1484–1490. 2006. View Article : Google Scholar : PubMed/NCBI
14	Nauta AJ, Westerhuis G, Kruisselbrink AB, Lurvink EG, Willemze R and Fibbe WE: Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting. Blood. 108:2114–2120. 2006. View Article : Google Scholar : PubMed/NCBI
15	Li Y and Lin F: Mesenchymal stem cells are injured by complement after their contact with serum. Blood. 120:3436–3443. 2012. View Article : Google Scholar : PubMed/NCBI
16	Blasius AL and Beutler B: Intracellular toll-like receptors. Immunity. 32:305–315. 2010. View Article : Google Scholar : PubMed/NCBI
17	Chen JC, Ng MM and Chu JJ: Activation of TLR2 and TLR6 by Dengue NS1 protein and its implications in the immunopathogenesis of Dengue virus infection. PLoS Pathog. 11:e10050532015. View Article : Google Scholar : PubMed/NCBI
18	Fu YS, Cheng YC, Lin MY, Cheng H, Chu PM, Chou SC, Shih YH, Ko MH and Sung MS: Conversion of human umbilical cord mesenchymal stem cells in Wharton's jelly to dopaminergic neurons in vitro: Potential therapeutic application for parkinsonism. Stem Cells. 24:115–124. 2006. View Article : Google Scholar : PubMed/NCBI
19	Tolar J, Le Blanc K, Keating A and Blazar BR: Concise review: Hitting the right spot with mesenchymal stromal cells. Stem Cells. 28:1446–1455. 2011. View Article : Google Scholar
20	Akira S, Uematsu S and Takeuchi O: Pathogen recognition and innate immunity. Cell. 124:783–801. 2006. View Article : Google Scholar : PubMed/NCBI
21	DelaRosa O and Lombardo E: Modulation of adult mesenchymal stem cells activity by toll-like receptors: Implications on therapeutic potential. Mediators Inflamm. 2010:8656012010. View Article : Google Scholar : PubMed/NCBI
22	Zhang L, Liu D, Pu D, Wang YW, Li L, He YQ, Li YL, Li L and Li WM: The TLR7 agonist Imiquimod promote the immunogenicity of mesenchymal stem cessl. Biol Res. 48:62015. View Article : Google Scholar : PubMed/NCBI
23	Zhang L, Liu D, Pu D, Wang YW, Li L, He Y, Li YL, Li L, Qiu ZC, Zhao S and Li WM: The role of Toll-like receptor 3 and 4 in regulating the function of mesenchymal stem cells isolated from umbilical cord. Int J Mol Med. 35:1003–1010. 2015. View Article : Google Scholar : PubMed/NCBI
24	Yang Y, Wang Y, Li L, Bao J, Chen F and Zhang L: Toll-like receptor 9 agonist stimulation enables osteogenic differentiation without altering the immune status of umbilical cord mesenchymal stem cells. Mol Med Rep. 12:8077–8084. 2015. View Article : Google Scholar : PubMed/NCBI
25	Huang XP, Sun Z, Miyagi Y, McDonald Kinkaid H, Zhang L, Weisel RD and Li RK: Differentiation of allogeneic mesenchymal stem cells induces immunogenicity and limits their long-term benefits for myocardial repair. Circulation. 122:2419–2429. 2010. View Article : Google Scholar : PubMed/NCBI