Open Access Peer-reviewed Research Article

SOCS1/2 controls NF-κB activity induced by HSP70 by degrading MyD88-adapter-like protein (Mal) in porcine macrophages

Main Article Content

Yanhong Yong
Lianyun Wu
Minglong Ma
Biao Fang
Tianyue Yu
Junyu Li
Canying Hu
Rumin Jia
Xianghong Ju corresponding author


Heat stress induces suppressor of cytokine signaling (SOCS) 1 and SOCS2 expression in the intestinal gut and disrupts inflammatory cytokine production in pigs. These changes may be important to the development of inflammatory bowel disease in heat-stressed pigs. However, the underlying mechanisms have not yet been completely elucidated. In the present study, we examined the roles of SOCS1 and SOCS2 in regulating the nuclear factor (NF)-κB pathway in CRL-2845 porcine macrophages. Ectopic expression of HSP70 significantly modulated NF-κB activity (p ≤ 0.05). Moreover, co-expression of SOCS1 or SOCS2 with HSP70 reduced NF-κB activity, which was abolished by SOCS1 or SOCS2 knockdown with  small interfering RNA. Additionally, myeloid differentiation factor 88 (MyD88)-adaptor-like (Mal) protein was down-regulated in cells expressing SOCS1 and SOCS2. SOCS1 and SOCS2 were found to negatively regulate the activity of NF-κB induced by HSP70 overexpression by degrading Mal. These findings may facilitate the development of novel SOCS1-based and SOCS2-based therapeutic strategies for controlling heat stress-related disorders in pigs.

heat stress, porcine macrophages, suppressor of cytokine signaling, nuclear factor-κB, MyD88-adapter-like protein

Article Details

How to Cite
Yong, Y., Wu, L., Ma, M., Fang, B., Yu, T., Li, J., Hu, C., Jia, R., & Ju, X. (2019). SOCS1/2 controls NF-κB activity induced by HSP70 by degrading MyD88-adapter-like protein (Mal) in porcine macrophages. Frontiers in Molecular Immunology, 1(1), 3-12.


  1. Ju XH, Yong YH, Xu HJ, et al. Impacts of heat stress on baseline immune measures and a subset of T cells in Bama miniature pigs. Livestock Science, 2011, 135(2): 289-292.
  2. Ju XH, Xu HJ, Yong YH, et al. Heat stress upregulation of Toll-like receptors 2/4 and acute inflammatory cytokines in peripheral blood mononuclear cell (PBMC) of Bama miniature pigs: an in vivo and in vitro study. animal, 2014, 8(9): 1462-1468.
  3. Tissi`eres A, Mitchell HK and Tracy UM. Protein synthesis in salivary glands of Drosophila melanogaster: relation to chromosome puffs. Journal of Molecular Biology, 1974, 84(3): 389.
  4. Schlesinger MJ. Heat shock proteins. Journal of Biological Chemistry, 1990, 21(21): 12111-12114.
  5. Chang YS, Lo CW, Sun FC, et al. Differential expression of Hsp90 isoforms in geldanamycin-treated 9L cells. Biochemical & Biophysical Research Communications, 2006, 344(1): 37-44.
  6. Borges TJ, Lotte W, Van HMJC, et al. The antiinflammatory mechanisms of Hsp70. Frontiers in Immunology, 2012, 3: 95.
  7. Hilton DJ, Richardson RT, Alexander WS, et al. Twenty proteins containing a C-terminal SOCS box form five structural classes. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(1): 114- 119.
  8. Linossi EM and Nicholson SE. Kinase inhibition, competitive binding and proteasomal degradation: resolving the molecular function of the suppressor of cytokine signaling (SOCS) proteins. Immunological Reviews, 2015, 266(1): 123-133.
  9. Narazaki M, Fujimoto M, Matsumoto T, et al. Three distinct domains of SSI-1/SOCS-1/JAB protein are required for its suppression of interleukin 6 signaling. Proceedings of the National Academy of Sciences, 1998, 95(22):13130-13134.
  10. Yasukawa H, Misawa H, Sakamoto H, et al. The JAKbinding protein JAB inhibits Janus tyrosine kinase activity through binding in the activation loop
  11. [D]. Rennes 1, 1999.
  12. Nicholson SE,Willson TA, Farley A, et al. Mutational analyses of the SOCS proteins suggest a dual domain requirement but distinct mechanisms for inhibition of LIF and IL-6 signal transduction. The EMBO Journal, 1999, 18(2): 375- 385.
  13. Waiboci LW, Ahmed CM, Mujtaba MG, et al. Both the suppressor of cytokine signaling 1 (SOCS-1) kinase inhibitory region and SOCS-1 mimetic bind to JAK2 autophosphorylation site: implications for the development of a SOCS-1 antagonist. Journal of Immunology, 2007, 178(8): 5058-5068.
  14. Mansell A, Smith R, Doyle S, et al. Suppressor of cytokine signaling 1 negatively regulates Toll-like receptor signaling by mediating Mal degradation. Nature Immunology, 2006, 7(2): 148-155.
  15. Tannahill GM, Joanne E, Barry AC, et al. SOCS2 can enhance interleukin-2 (IL-2) and IL-3 signaling by accelerating SOCS3 degradation. Molecular & Cellular Biology, 2005, 25(20): 9115-9126.
  16. Elizabeth RB, Amilcar FM and Leandro FP. Suppressor of cytokine signaling (SOCS) 2, a protein with multiple functions. Cytokine & Growth Factor Reviews, 2006, 17(6): 431-439.
  17. Hansen PJ and Ar´echiga CF. Strategies for managing reproduction in the heat-stressed dairy cow. Journal of Animal Science, 1999, 77(Suppl 2): 36-50. 236x
  18. Cortez MB, Gonzalez RMS, Nathaniel S, et al. SOCS2- induced proteasome-dependent TRAF6 degradation: a common anti-inflammatory pathway for control of innate immune responses. Plos One, 2012, 7(6): e38384.
  19. Bruel T, Guibon R, Melo S, et al. Epithelial induction of porcine suppressor of cytokine signaling 2 (SOCS2) gene expression in response to Entamoeba histolytica. Developmental & Comparative Immunology, 2010, 34(5): 562-571.
  20. Gernot P, Harald S, Albert D, et al. Suppressor of cytokine signaling 2 is a feedback inhibitor of TLR-induced activation in human monocyte-derived dendritic cells. Journal of Immunology, 2011, 187(6): 2875-2884.
  21. Sugimoto N, Oida T, Hirota K, et al. Foxp3-dependent and -independent molecules specific for CD25+CD4+ natural regulatory T cells revealed by DNA microarray analysis. International Immunology, 2007, 18(7): 375-377.
  22. Haribhai D, Lin W, Edwards B, et al. A Central Role for Induced Regulatory T Cells in Tolerance Induction in Experimental Colitis. Journal of Immunology, 2009, 182(6): 3461.
  23. Ju XH, Yong YH, Xu HJ, et al. Selection of reference genes for gene expression studies in PBMC from Bama miniature pig under heat stress. Veterinary Immunology & Immunopathology, 2011, 144(1): 160-166.
  24. Yong YH, Wang P, Jia RM, et al. SOCS3 control the activity of NF-B induced by HSP70 via degradation of MyD88- adapter-like protein (Mal) in IPEC-J2 cells. International Journal of Hyperthermia, 2019, 36(1): 151-159.
  25. Zhu J, Lai K, Brownile R, et al. Porcine TLR8 and TLR7 are both activated by a selective TLR7 ligand, imiquimod. Molecular immunology, 2008, 45(11): 3238-3243.
  26. An H, Xu H, Yu Y, et al. Up-regulation of TLR9 gene expression by LPS in mouse macrophages via activation of NF-B, ERK and p38 MAPK signal pathways. Immunology letters, 2002, 81(3): 165-169.
  27. Bao B, Prasad A, Beck FW, et al. Toxic effect of zinc on NFkappaB, IL-2, IL-2 receptor alpha, and TNF-alpha in HUT- 78 (Th(0)) cells. Toxicology Letters, 2006, 166(3): 222.
  28. Craig EA and Gross CA. Is hsp70 the cellular thermometer? Trends in Biochemical Sciences, 1991, 16(4): 135-140.
  29. Yu H, Bao ED, Zhao RQ, et al. Effect of transportation stress on heat shock protein 70 concentration and mRNA expression in heart and kidney tissues and serum enzyme activities and hormone concentrations of pigs. American Journal of Veterinary Research, 2007, 68(11): 1145-1150.
  30. Calderwood SK, Theriault JR and Gong J. How is the immune response affected by hyperthermia and heat shock proteins?. International Journal of Hyperthermia, 2005, 21(8): 713-716.
  31. Senf SM, Dodd SL, Mcclung JM, et al. Hsp70 overexpression inhibits NF-κB and Foxo3a transcriptional activities and prevents skeletal muscle atrophy. The FASEB Journal, 2008, 22(11): 3836-3845.
  32. Alexander WS, Starr R, Fenner JE, et al. SOCS1 Is a Critical Inhibitor of Interferon Signaling and Prevents the Potentially Fatal Neonatal Actions of this Cytokine. Cell, 1999, 98(5): 597-608.
  33. Yoshikawa H, Matsubara K, Qian GS, et al. SOCS-1, a negative regulator of the JAK/STAT pathway, is silenced by methylation in human hepatocellular carcinoma and shows growth-suppression activity. Nature Genetics, 2001, 28(1): 29-35.
  34. Tamiya T, Kashiwagi I, Takahashi R, et al. Suppressors of cytokine signaling (SOCS) proteins and JAK/STAT pathways: regulation of T-cell inflammation by SOCS1 and SOCS3. Arteriosclerosis Thrombosis and Vascular Biology, 2011, 31(5): 980-985.
  35. Ryo A, Suizu F, Yoshida Y, et al. Regulation of NF- κB Signaling by Pin1-Dependent Prolyl Isomerization and Ubiquitin-Mediated Proteolysis of p65/RelA. Molecular Cell, 2003, 12(6): 1413-1426.
  36. Bruce B. Inferences, questions and possibilities in Toll-like receptor signalling. Nature, 2004, 430(6996): 257-263.
  37. Karin M and Greten FR. NF-κB: linking inflammation and immunity to cancer development and progression. Nature Reviews Immunology, 2005, 5(10): 749-759.
  38. Kawai T and Akira S. Signaling to NF-κB by Toll-like receptors. Trends in Molecular Medicine, 2007, 13(11): 460- 469.
  39. Danielle M. Role of NF-κB in beta-cell death. Biochemical Society Transactions, 2008, 36(3): 334-339.
  40. Shisler JL and Jin XL. The vaccinia virus K1L gene product inhibits host NF-κB activation by preventing IκBa degradation. Journal of virology, 2004, 78(7): 3553-3560.
  41. Inoue H, Arai Y, Terauchi R, et al. Effect of inflammation stress on the hypertrophic differentiation related gene expression in cultured chondrocytes. Osteoarthritis and Cartilage. Osteoarthritis and Cartilage, 2014, 22: S294.
  42. Amaral BCD, Connor EE, Tao S, et al. Heat stress abatement during the dry period influences prolactin signaling in lymphocytes. Domestic Animal Endocrinology, 2010, 38(1): 38-45.
  43. Machado FS, Johndrow JE, Esper L, et al. Antiinflammatory actions of lipoxin A4 and aspirin-triggered lipoxin are SOCS-2 dependent. Nature Medicine, 2006, 12(3): 330-334.
  44. Lee JS, Paek NS, Kwon OS, et al. Anti-inflammatory actions of probiotics through activating suppressor of cytokine signaling (SOCS) expression and signaling in Helicobacter pylori infection: A novel mechanism. Journal of Gastroenterology and Hepatology, 2010, 25(1): 194-202.
  45. Buford TW, Cooke MB and Willoughby DS. Resistance exercise-induced changes of inflammatory gene expression within human skeletal muscle. European Journal of Applied Physiology, 2009, 107(4): 463-471.
  46. Wilson HM. SOCS Proteins in Macrophage Polarization and Function. Frontiers in Immunology, 2014, 5(5): 357.
  47. Bonjardim CA, Ferreira PCP and Kroon EG. Interferons: signaling, antiviral and viral evasion. Immunology Letters, 2009, 122(1): 1-11.
  48. Miggin SM, Palsson-Mcdermott E, Dunne A, et al. NF-κB activation by the Toll-IL-1 receptor domain protein MyD88 adapter-like is regulated by caspase-1. Proceedings of the National Academy of Sciences, 2007, 104(9): 3372-3377.
  49. Tagami N, Serada S, Fujimoto M, et al. Abstract 1348: Suppressor of cytokine signaling (SOCS)-1 suppresses a proliferation of malignant melanoma cells via the suppression of JAK/STAT and the activation of p53 signaling pathways. Cancer Research, 2014, 74(19 Suppl): 1348.
  50. Recio C, Oguiza A, Mallavia B, et al. Gene delivery of suppressors of cytokine signaling (SOCS) inhibits inflammation and atherosclerosis development in mice. Basic Research in Cardiology, 2015, 110(2): 8.
  51. Hu J, Lou D, Carow B, et al. LPS regulates SOCS2 transcription in a type I interferon dependent autocrineparacrine loop. Plos One, 2012, 7(1): e30166.