Association between TLR2 and TLR4 Expression and Response to Induction Therapy in Acute Myeloid Leukemia Patients

  • Mani Ramzi Hematology Research Center, Department of Hematology, Medical Oncology and Stem Cell Transplantation, Shiraz University of Medical Sciences, Shiraz, Iran
  • Abolfazl Khalafi-Nezhad Hematology Research Center, Department of Hematology, Medical Oncology and Stem Cell Transplantation, Shiraz University of Medical Sciences, Shiraz, Iran
  • Mahdiyar Iravani Saadi Hematology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
  • Zahra Jowkar Department of Operative Dentistry, School of Dentistry, Shiraz University of Medical Sciences, Shiraz, Iran
Keywords: Toll-like receptors, Induction chemotherapy, Acute myeloid leukemia


Background: Toll-like receptors (TLRs) are a family of transmembrane pattern-recognition receptors that play a crucial role in the realization of innate and adaptive immune response. TLRs may play a role in tumor development and growth because of expression or up-regulation of functional TLRs in some tumors and tumor cell lines. The participation of TLRs in the pathogenesis of acute myeloid leukemia (AML) remains unspecified. This study aimed to investigate the effect of TLR2 and TLR4 expression in peripheral blood mononuclear cells of AML patients in response to induction chemotherapy. Materials and Methods: Eighty- five patients with newly diagnosed AML were evaluated. Using quantitative reverse transcriptase PCR, the mRNA expression of genes TLR2 and TLR4 was measured before starting and after induction chemotherapy. The differences in the mean expression levels of TLR2 and TLR4 before and after chemotherapy were compared using a paired t-test. The mean expression levels of TLR2 and TLR4 regarding laboratory data were analyzed by one-way ANOVA and Chi-square test. Results: We found that the mRNA expression of TLR2 after induction chemotherapy was significantly lower as compared to before treatment (p=0.001). Also, we found a lower TLR4 gene expression level after chemotherapy as compared to before chemotherapy, albeit it was not statistically significant (p=0.21). Moreover, we observed significantly higher expression of TLR2 and TLR4 in AML-M3 cases compared to non-M3 AML patients. Conclusion: The decreased expression of TLR4 in leukemic samples after induction chemotherapy might indicate a novel potential prognostic role for this receptor, particularly in AML-M3 cases.


1. Estey EH. General approach to, and perspectives on clinical research in, older patients with newly diagnosed acute myeloid leukemia. Semin Hematol. 2006;43(2):89-95.
2. Döhner H, Estey EH, Amadori S, et al. Diagnosis and management of acute myeloid leukemia in adults: recommendations from an international expert panel, on behalf of the European LeukemiaNet. Blood. 2010;115(3):453-74.
3. Ridgway D. The first 1000 dendritic cell vaccinees. Cancer Invest. 2003;21(6):873-86.
4. Hagenbeek A, Gadeberg O, Johnson P, et al. First clinical use of ofatumumab, a novel fully human anti-CD20 monoclonal antibody in relapsed or refractory follicular lymphoma: results of a phase 1/2 trial. Blood. 2008;111(12):5486-95.
5. Mavromatis B, Cheson BD. Monoclonal antibody therapy of chronic lymphocytic leukemia. J Clin Oncol. 2003;21(9):1874-81.
6. Ibrahem L, Mahfouz R, Elhelw L, et al. Prognostic significance of DNMT3A mutations in patients with acute myeloid leukemia. Blood Cells Mol Dis. 2015;54(1):84-9.
7. Li K, Lv XX, Hua F, Lin H, et al. Targeting acute myeloid leukemia with a proapoptotic peptide conjugated to a Toll-like receptor 2-mediated cell-penetrating peptide. Int J Cancer. 2014;134(3):692-702.
8. Nasrollahi SA, Taghibiglou C, Azizi E, et al. Cell-penetrating peptides as a novel transdermal drug delivery system. Chem Biol Drug Des. 2012;80(5):639-46.
9. Ignatz-Hoover JJ, Wang H, Moreton SA, et al. The role of TLR8 signaling in acute myeloid leukemia differentiation. Leukemia. 2015;29(4):918-926.
10. Rybka J, Butrym A, Wróbel T , et al. The Expression of Toll-Like Receptors in Patients with B-Cell Chronic Lymphocytic Leukemia. Arch Immunol Ther Exp (Warsz). 2016;64(Suppl 1):147-150.
11. Harsini S, Beigy M, Akhavan-Sabbagh M, et al. Toll-like receptors in lymphoid malignancies: double-edged sword. Crit Rev Oncol Hematol. 2014;89(2):262-83.
12. Wang RF, Miyahara Y, Wang HY. Toll-like receptors and immune regulation: implications for cancer therapy. Oncogene. 2008;27(2):181-9.
13. Basith S, Manavalan B, Yoo TH, et al. Roles of toll-like receptors in cancer: a double-edged sword for defense and offense. Arch Pharm Res. 2012;35(8):1297-316.
14. Harberts E, Gaspari AA. TLR signaling and DNA repair: are they associated? J Invest Dermatol. 2013;133(2):296-302.
15. Zhu L, Yuan H, Jiang T, et al. Association of TLR2 and TLR4 polymorphisms with risk of cancer: A meta-analysis. PloS one. 2013;8(12):e82858.
16. Collins SJ, Ruscetti FW, Gallagher RE, et al. Normal functional characteristics of cultured human promyelocytic leukemia cells (HL-60) after induction of differentiation by dimethylsulfoxide. J Exp Med. 1979; 149(4): 969–974.
17. Bohnhorst J, Rasmussen T, Moen SH, et al. Toll-like receptors mediate proliferation and survival of multiple myeloma cells. Leukemia. 2006;20(6):1138-44.
18. Spaner DE, Masellis A. Toll-like receptor agonists in the treatment of chronic lymphocytic leukemia. Leukemia. 2007;21(1):53-60.
19. Okamoto M, Hirai H, Taniguchi K, et al. Toll-like receptors (TLRs) are expressed by myeloid leukaemia cell lines, but fail to trigger differentiation in response to the respective TLR ligands. Br J Haematol. 2009;147(4):585-7.
20. Bennett JM, Catovsky D, Daniel MT, et al. Proposed revised criteria for the classification of acute myeloid leukemia. A report of the French-American-British Cooperative Group. Ann Intern Med. 1985;103(4):620-5.
21. de Greef GE, van Putten WL, Boogaerts M, et al. Criteria for defining a complete remission in acute myeloid leukaemia revisited. An analysis of patients treated in HOVON-SAKK co-operative group studies. Br J Haematol. 2005;128(2):184-91.
22. Schmohl JU, Nuebling T, Wild J, et al. Expression of RANK-L and in part of PD-1 on blasts in patients with acute myeloid leukemia correlates with prognosis. Eur J Haematol. 2016;97(6):517-527.
23. Salomao R, Brunialti MK, Rapozo MM, et al. Bacterial sensing, cell signaling, and modulation of the immune response during sepsis. Shock. 2012;38(3):227-42.
24. Rybka J, Butrym A, Wróbel T , et al. The expression of Toll-like receptors in patients with acute myeloid leukemia treated with induction chemotherapy. Leuk Res. 2015;39(3):318-22.
25. Sánchez-Cuaxospa M, Contreras-Ramos A, Pérez-Figueroa E, et al. Low expression of Toll-like receptors in peripheral blood mononuclear cells of pediatric patients with acute lymphoblastic leukemia. Int J Oncol. 2016;49(2):675-81.
26. Renshaw M, Rockwell J, Engleman C, et al. Cutting edge: impaired Toll-like receptor expression and function in aging. J Immunol. 2002;169(9):4697-701.
27. Härter L, Mica L, Stocker R, et al. Increased expression of toll-like receptor-2 and-4 on leukocytes from patients with sepsis. Shock. 2004;22(5):403-9.
28. Armstrong L, Medford A, Hunter K, et al. Differential expression of Toll‐like receptor (TLR)‐2 and TLR‐4 on monocytes in human sepsis. Clin Exp Immunol. 2004;136(2):312-9.
29. Nagai Y, Garrett KP, Ohta S, et al. Toll-like receptors on hematopoietic progenitor cells stimulate innate immune system replenishment. Immunity. 2006;24(6):801-12.
30. Sioud M, Fløisand Y. TLR agonists induce the differentiation of human bone marrow CD34+ progenitors into CD11c+ CD80/86+ DC capable of inducing a Th1-type response. Eur J Immunol. 2007;37(10):2834-46.
31. Wald DN, Vermaat HM, Zang S, et al. Identification of 6-benzylthioinosine as a myeloid leukemia differentiation-inducing compound. Cancer Res. 2008;68(11):4369-4376.
32. Sioud M, Fløisand Y, Forfang L, et al. Signaling through toll-like receptor 7/8 induces the differentiation of human bone marrow CD34+ progenitor cells along the myeloid lineage. J Mol Biol. 2006;364(5):945-54.
33. Wang Z, Smith KS, Murphy M, et al. Glycogen synthase kinase 3 in MLL leukaemia maintenance and targeted therapy. Nature. 2008;455(7217):1205-9.
34. Beck B, Dörfel D, Lichtenegger FS, et al. Effects of TLR agonists on maturation and function of 3-day dendritic cells from AML patients in complete remission. J Transl Med. 2011;9:151.
35. Purdue MP, Lan Q, Wang SS, et al. A pooled investigation of Toll-like receptor gene variants and risk of non-Hodgkin lymphoma. Carcinogenesis. 2009;30(2):275-81.
36. Lazarus R, Raby BA, Lange C, et al. TOLL-like receptor 10 genetic variation is associated with asthma in two independent samples. Am J Respir Crit Care Med. 2004;170(6):594-600.
37. Kesh S, Mensah NY, Peterlongo P, et al. TLR1 and TLR6 polymorphisms are associated with susceptibility to invasive aspergillosis after allogeneic stem cell transplantation. Ann N Y Acad Sci. 2005;1062:95-103.
38. Nieters A, Beckmann L, Deeg E, et al. Gene polymorphisms in Toll-like receptors, interleukin-10, and interleukin-10 receptor alpha and lymphoma risk. Genes Immun. 2006;7(8):615-24.
39. Torok AM, Bouton AH, Goldberg JB. Helicobacter pylori induces interleukin-8 secretion by Toll-like receptor 2- and Toll-like receptor 5-dependent and -independent pathways. Infect Immun. 2005;73(3):1523-31.
40. Webb RN, Cruse JM, Lewis RE. Decreased TLR4 gene expression in leukemic leukocyte populations. Exp Mol Pathol. 2009;87(2):117-26.
41. Huang B, Zhao J, Shen S, et al. Listeria monocytogenes promotes tumor growth via tumor cell toll-like receptor 2 signaling. Cancer Res. 2007;67(9):4346-52.
42. Chen R, Alvero AB, Silasi DA, et al. Cancers take their Toll--the function and regulation of Toll-like receptors in cancer cells. Oncogene. 2008;27(2):225-33.
43. He W, Liu Q, Wang L, et al. TLR4 signaling promotes immune escape of human lung cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Mol Immunol. 2007;44(11):2850-9.
44. Wang JH, Manning BJ, Wu QD, et al. Endotoxin/lipopolysaccharide activates NF-kappa B and enhances tumor cell adhesion and invasion through a beta 1 integrin-dependent mechanism. J Immunol. 2003;170(2):795-804.
45. Ahmed A, Wang JH, Redmond HP. Silencing of TLR4 increases tumor progression and lung metastasis in a murine model of breast cancer. Ann Surg Oncol. 2013;20 Suppl 3:S389-96.
How to Cite
Ramzi M, Khalafi-Nezhad A, Iravani Saadi M, Jowkar Z. Association between TLR2 and TLR4 Expression and Response to Induction Therapy in Acute Myeloid Leukemia Patients. ijhoscr. 12(4):302-11.
Original Article(s)