Noninvasive imaging of hepatocyte IL-6/STAT3 signaling pathway for evaluating inflammation responses induced by end-stage stored whole blood transfusion

  • Zhengjun Wang
  • Yulong Zhang
  • Qianqian Zhou
  • Ping Ma
  • Xiaohui WangEmail author
  • Linsheng ZhanEmail author
Original Research Paper



To monitor the inflammatory storage lesions of end-stage stored whole blood (SWB) using a noninvasive STAT3 signal pathway mouse model.


In this study, we present a hydrodynamic transfection-based STAT3-Luc mouse model in which hepatocyte STAT3 signal pathway activation can be monitored by measuring luciferase activity using a noninvasive imaging system. Such a mouse model may reflect systemic IL-6 and inflammation levels by monitoring the activation of STAT3. During end-stage SWB transfusion, in vivo imaging of STAT3-Luc mice showed obvious luciferase activity in the hepatic region, which was consistent with an increase in IL-6 levels in the liver homogenate and circulation. We also confirmed that the mononuclear phagocytic system contributed to the elevation of serum and liver IL-6 after end-stage SWB transfusion.


The hepatocyte STAT3 signaling pathway, which is activated by end-stage SWB transfusion, is associated with the elevation of systemic IL-6 secreted by macrophages. The STAT3-Luc mouse may serve as a mouse model for monitoring inflammation responses after end-stage SWB transfusion.


Whole blood Hydrodynamic transfection Bioluminescence imaging IL-6/STAT3 Inflammatory 



This work was supported by Foundation of AHJ17J002-06, the National Natural Science Foundation of China (81800183) and the Mega-Project of Science Research (2017ZX10304402-003-004, 2017ZX10304402-003-011).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Badr CE, Tannous BA (2011) Bioluminescence imaging: progress and applications. Trends Biotechnol 29:624–633CrossRefGoogle Scholar
  2. Contag CH, Jenkins D, Contag PR, Negrin RS (2000) Use of reporter genes for optical measurements of neoplastic disease in vivo. Neoplasia 2:41–52CrossRefGoogle Scholar
  3. Daniel Y, Sailliol A, Pouget T, Peyrefitte S, Ausset S, Martinaud C (2017) Whole blood transfusion closest to the point-of-injury during French remote military operations. J Trauma Acute Care Surg 82:1CrossRefGoogle Scholar
  4. Elster EA, Bailey J (2017) Prehospital blood transfusion for combat casualties. JAMA 318:1548–1549CrossRefGoogle Scholar
  5. Emery P et al (2008) IL-6 receptor inhibition with tocilizumab improves treatment outcomes in patients with rheumatoid arthritis refractory to anti-tumour necrosis factor biologicals: results from a 24-week multicentre randomised placebo-controlled trial. Ann Rheum Dis 67:1516–1523CrossRefGoogle Scholar
  6. Gang N et al (2013) Longitudinal bioluminescence imaging of the dynamics of Doxorubicin induced apoptosis. Theranostics 3:190–200CrossRefGoogle Scholar
  7. Garcíaroa M et al (2017) Red blood cell storage time and transfusion: current practice, concerns and future perspectives. Blood Transfus 15:222Google Scholar
  8. Greenbaum BH (1991) Transfusion-associated graft-versus-host disease: historical perspectives, incidence, and current use of irradiated blood products. J Clin Oncol 9:1889–1902CrossRefGoogle Scholar
  9. Gurney JM, Spinella PC (2018) Blood transfusion management in the severely bleeding military patient. Curr Opin Anaesthesiol 31:207Google Scholar
  10. Harvey AR, Basavaraju SV, Chung KW, Kuehnert MJ (2015) Transfusion-related adverse reactions reported to the National Healthcare Safety Network Hemovigilance Module, United States, 2010 to 2012. Transfusion 55:709–718CrossRefGoogle Scholar
  11. Hergenroeder GW, Moore AN, McCoy JP, Samsel L, Iii NHW, Clifton GL, Dash PK (2010) Serum IL-6: a candidate biomarker for intracranial pressure elevation following isolated traumatic brain injury. J Neuroinflamm 7(1):19CrossRefGoogle Scholar
  12. Hess JR, Greenwalt TG (2002) Storage of red blood cells: new approaches. Transfus Med Rev 16:283–295CrossRefGoogle Scholar
  13. Higuchi N et al (2003) Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar-Kyoto rats. Gene Therapy 10:1297–1310CrossRefGoogle Scholar
  14. Hod EA et al (2010) Transfusion of red blood cells after prolonged storage produces harmful effects that are mediated by iron and inflammation. Blood 115:4284–4292CrossRefGoogle Scholar
  15. Holcomb JB, Jenkins DH (2018) Get ready: whole blood is back and it’s good for patients. Transfusion 58:1821–1823CrossRefGoogle Scholar
  16. Knutson M, Wessling-Resnick M (2003) Iron Metabolism in the Reticuloendothelial System Crc. Crit Rev Biochem 38:61–88CrossRefGoogle Scholar
  17. Nam-Hoon K, Mun-Yong L, Seon-Joo P, Jeong-Sun C, Mi-Kyung O, In-Sook K (2010) Auranofin blocks interleukin-6 signalling by inhibiting phosphorylation of JAK1 and STAT3. Immunology 122:607–614Google Scholar
  18. Ning Z, Aneil W, Bonnie L, Richard L, Contag PR, Purchio AF, West DB (2003) An inducible nitric oxide synthase-luciferase reporter system for in vivo testing of anti-inflammatory compounds in transgenic mice. J Immunol 170:6307CrossRefGoogle Scholar
  19. Ning Z, Ahsan MH, Purchio AF, West DB (2005) Serum amyloid A-luciferase transgenic mice: response to sepsis, acute arthritis, and contact hypersensitivity and the effects of proteasome inhibition. J Immunol 174:8125–8134CrossRefGoogle Scholar
  20. Park E et al (2010) Dietary and genetic obesity promote liver inflammation and tumorigenesis by enhancing IL-6 and TNF. Expr Cell 140:197–208CrossRefGoogle Scholar
  21. Shackelford SA et al (2017) Association of prehospital blood product transfusion during medical evacuation of combat casualties in afghanistan with acute and 30-day survival. JAMA 318:1581CrossRefGoogle Scholar
  22. Theurl I et al (2016) On-demand erythrocyte disposal and iron recycling requires transient macrophages in the liver. Nat Med 22(8):945–951CrossRefGoogle Scholar
  23. Vlaar APJ, Juffermans NP (2013) Transfusion-related acute lung injury: a clinical review. Lancet 382:984–994CrossRefGoogle Scholar
  24. Woodard HQ, White DR (1986) The composition of body tissues. Br J Radiol 59:1209CrossRefGoogle Scholar
  25. Wu T et al (2017) Noninvasive imaging of stored red blood cell-transfusion aggravating sepsis-induced liver injury associated with increased activation of M1-polarized Kupffer cells. Shock 48(4):459–466CrossRefGoogle Scholar
  26. Xia S et al (2016) Sodium pyruvate improves the storage damage of red blood cells Chinese. J Blood Transfus 29:353–356Google Scholar
  27. Yan S et al (2013) Establishment of stable reporter expression for in vivo imaging of nuclear factor-κB activation in mouse liver. Theranostics 3:841–850CrossRefGoogle Scholar
  28. Yazer MH, Cap AP, Spinella PC, Alarcon L, Triulzi DJ (2018) How do I implement a whole blood program for massively bleeding patients? Transfusion 58(3):622–628CrossRefGoogle Scholar
  29. Yong Z et al (2012) Noninvasive molecular imaging of interferon beta activation in mouse liver. Liver Int 32:383–391Google Scholar
  30. Zhang W, Feng JQ, Harris SE, Contag PR, Stevenson DK, Contag CH (2001) Rapid in vivo functional analysis of transgenes in mice using whole body imaging of luciferase expression. Transgenic Res 10:423–434CrossRefGoogle Scholar
  31. Zhang Y et al (2016) Non-invasive imaging serum amyloid A activation through the NF-κB signal pathway upon gold nanostructure exposure. Small 12:3270–3282CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute of Health Service and Transfusion MedicineBeijingPeople’s Republic of China

Personalised recommendations