Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
5-Year Impact Factor – 2.2
Scopus CiteScore – 3.4 (CiteScore Tracker 3.4)
Index Copernicus  – 161.11; MEiN – 140 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

Download original text (EN)

Advances in Clinical and Experimental Medicine

2020, vol. 29, nr 7, July, p. 833–840

doi: 10.17219/acem/121924

Publication type: original article

Language: English

License: Creative Commons Attribution 3.0 Unported (CC BY 3.0)

Download citation:

  • BIBTEX (JabRef, Mendeley)
  • RIS (Papers, Reference Manager, RefWorks, Zotero)

Autologous transfusion of “old” red blood cells-induced M2 macrophage polarization through IL-10-Nrf2-HO-1 signaling complexes

Zhen-Zhou Li1,2,A,B,C,D,E, Zi-Wei Zhang3,A,B,C,D, Huan Wang2,B,C,E,F, Yong-Quan Chen2,B,C,E, Xiao-Fang Zhou2,B,C, Li-Shuang Duan2,C,E, Xiao-Xiao Wang2,E, Feng Xu4,E, Jian-Rong Guo1,2,B,F

1 Shanghai Pudong New Area Gongli Hospital Training Base, Ningxia Medical University, China

2 Department of Anesthesiology, Gongli Hospital, Second Military Medical University, Shanghai, China

3 Department of Anesthesiology, Yijishan Hospital, Wannan Medical College, Wuhu, China

4 Department of Anesthesiology, Anhui Province No. 2 People’s Hospital, Hefei, China


Background. Red blood cell (RBC) transfusion is associated with systemic inflammation and immune suppression as adverse outcomes.
Objectives. To investigate the immunomodulatory function of the transfused autologous RBC in altering pro-inflammatory and immunosuppressive effects.
Material and Methods. A total of 24 Sprague Dawley male rats were randomly divided into 3 groups (n = 8 in each group). Group 1 did not receive blood transfusions, while the other 2 groups of rats separately received transfusion of RBC stored for 14 days (group 2) and 35 days (group 3). The rats were treated with HO-1 inhibitor, HO-1 inducer and nuclear factor erythroid 2-related factor 2 (Nrf2) activator after they separately received autologous transfusion of RBC that were cryopreserved for 14 days or 35 days. The blood samples of the rats were collected 12 h after the transfusion, and the macrophage phenotype of M1 and M2 were analyzed with flow cytometry (FCM). Also, the surface protein expression of CD68 and CD200R in macrophages were analyzed and the inflammatory signals in the serum were measured with enzyme-linked immunosorbent assay (ELISA). Moreover, the location and expression of proteins heme oxygenase 1 (HO-1), arginine 1 (Arg-1) and nitric oxide synthase 2 (NOS2) in macrophage were detected with immunofluorescence (IF).
Results. Autologous transfusion of long-time stored (“old”) RBC promoted macrophage polarization to M2 phenotype and upregulated the expression of its surface proteins CD68 and CD200R. The pro-inflammatory cytokines tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-1β, and IL-18 were inhibited, and the secretion of NOS isoforms (iNOS) in serum was reduced with blood transfusion; contrarily, the production of IL-10 and CCL22 was increased. Additionally, HO-1, Arg-1 and NOS2 proteins were located in the cytoplasm, and HO-1 and Arg-1 proteins were highly expressed in macrophage, while the expression of protein NOS2 was low. Moreover, Nrf2, HO-1 and Arg-1 proteins were upregulated in macrophage after receiving “old” RBC transfusion.
Conclusion. Autologous transfusion of “old” RBC drove the macrophage phenotype toward M2 macrophages and induced immunosuppressive effects through the IL-10-NRF2-HO-1 signals.

Key words

immunosuppressive treatment, red blood cell transfusion, M2 macrophage, IL-10-NRF2-HO-1 signals

References (36)

  1. Carson JL, Guyatt G, Heddle NM, et al. Clinical practice guidelines from the AABB: Red blood cell transfusion thresholds and storage. JAMA. 2016;316(19):2025–2035.
  2. Dagur PK, McCoy JP. Collection, storage, and preparation of human blood cells. Curr Protoc Cytom. 2015;73:5.1.1–5.1.16. doi:10.1002/0471142956.cy0501s73
  3. Muszynski JA, Spinella PC, Cholette JM, et al; Pediatric Critical Care Blood Research Network (Blood Net). Transfusion-related immunomodulation: Review of the literature and implications for pediatric critical illness. Transfusion. 2017;57(1):195–206.
  4. Hod EA, Godbey EA. The outsider adverse event in transfusion: Inflammation. Presse Med. 2016;45(7–8 Pt 2):e325–e329.
  5. van de Watering L. Red cell storage and prognosis. Vox Sang. 2011;100(1):36–45.
  6. Karon BS, Hoyer JD, Stubbs JR, Thomas DD. Changes in Band 3 oligomeric state precede cell membrane phospholipid loss during blood bank storage of red blood cells. Transfusion. 2009;49(7):1435–1442.
  7. D’Amici GM, Rinalducci S, Zolla L. Proteomic analysis of RBC membrane protein degradation during blood storage. J Proteome Res. 2007;6(8):3242–3255.
  8. Ozment CP, Mamo LB, Campbell ML, Lokhnygina Y, Ghio AJ, Turi JL. Transfusion-related biologic effects and free hemoglobin, heme, and iron. Transfusion. 2013;53(4):732–740.
  9. Banerjee S. Glyoxal-induced modification enhances stability of hemoglobin and lowers iron-mediated oxidation reactions of the heme protein: An in vitro study. Int J Biol Macromol. 2018;107(Pt A):494–501.
  10. Spitalnik SL. Stored red blood cell transfusions: Iron, inflammation, immunity, and infection. Transfusion. 2014;54(10):2365–2371.
  11. Baek JH, Yalamanoglu A, Gao Y, et al. Iron accelerates hemoglobin oxidation increasing mortality in vascular diseased guinea pigs following transfusion of stored blood. JCI Insight. 2017;2(9):e93577.
  12. Dutra FF, Bozza MT. Heme on innate immunity and inflammation. Front Pharmacol. 2014;5:115.
  13. Haldar M, Kohyama M, Yick-Lun So A, et al. Heme-mediated SPI-C induction promotes monocyte differentiation into iron-recycling macrophages. Cell. 2014;156(6):1223–1234.
  14. Vinchi F, Costa da Silva M, Ingoglia G, et al. Hemopexin therapy reverts heme-induced pro-inflammatory phenotypic switching of macrophages in a mouse model of sickle cell disease. Blood. 2016;127(4):473–486.
  15. Straat M, van Hezel ME, Böing A, et al. Monocyte-mediated activation of endothelial cells occurs only after binding to extracellular vesicles from red blood cell products, a process mediated by β-integrin. Transfusion. 2016;56(12):3012–3020.
  16. Rubin O, Delobel J, Prudent M, et al. Red blood cell-derived microparticles isolated from blood units initiate and propagate thrombin generation. Transfusion. 2013;53(8):1744–1754.
  17. Straat M, Böing AN, Tuip-De Boer A, Nieuwland R, Juffermans NP. Extracellular vesicles from red blood cell products induce a strong pro-inflammatory host response, dependent on both numbers and storage duration. Transfus Med Hemother. 2016;43(4):302–305.
  18. Dinkla S, Novotný VMJ, Joosten I, Bosman GJCGM. Storage-induced changes in erythrocyte membrane proteins promote recognition by autoantibodies. PLoS One. 2012;7(8):e42250.
  19. Yazdanbakhsh K, Bao W, Zhong H. Immunoregulatory effects of stored red blood cells. Hematology Am Soc Hematol Educ Program. 2011;2011:466–469.
  20. Hod EA, Brittenham GM, Billote GB, et al. Transfusion of human volunteers with older, stored red blood cells produces extravascular hemolysis and circulating non-transferrin-bound iron. Blood. 2011;118(25):6675–6682.
  21. Hod EA, Zhang N, Sokol SA, et al. Transfusion of red blood cells after prolonged storage produces harmful effects that are mediated by iron and inflammation. Blood. 2010;115(21):4284–4292.
  22. Hult A, Malm C, Oldenborg PA. Transfusion of cryopreserved human red blood cells into healthy humans is associated with rapid extravascular hemolysis without a pro-inflammatory cytokine response. Transfusion. 2013;53(1):28–33.
  23. Knutson MD, Vafa MR, Haile DJ, Wessling-Resnick M. Iron loading and erythrophagocytosis increase ferroportin 1 (FPN1) expression in J774 macrophages. Blood. 2003;102(12):4191–4197.
  24. Youssef LA, Rebbaa A, Pampou S, et al. Increased erythrophagocytosis induces ferroptosis in red pulp macrophages in a mouse model of transfusion. Blood. 2018;131(23):2581–2593.
  25. Hao K, Hanawa H, Ding L, et al. Free heme is a danger signal inducing expression of pro-inflammatory proteins in cultured cells derived from normal rat hearts. Mol Immunol. 2011;48(9–10):1191–1202.
  26. Larsen R, Gouveia Z, Soares MP, Gozzelino R. Heme cytotoxicity and the pathogenesis of immune-mediated inflammatory diseases. Front Pharmacol. 2012;3:77.
  27. Cherry AD, Piantadosi CA. Regulation of mitochondrial biogenesis and its intersection with inflammatory responses. Antioxid Redox Signal. 2015;22(12):965–976.
  28. Naito Y, Takagi T, Higashimura Y. Heme oxygenase-1 and anti-inflammatory M2 macrophages. Arch Biochem Biophys. 2014;564:83–88.
  29. Weis N, Weigert A, von Knethen A, Brüne B, Luo K. Heme oxygenase-1 contributes to an alternative macrophage activation profile induced by apoptotic cell supernatants. Mol Biol Cell. 2009;20(5):1280–1288.
  30. Park EJ, Kim YM, Park SW, et al. Induction of HO-1 through p38 MAPK/Nrf2 signaling pathway by ethanol extract of Inula helenium L. reduces inflammation in LPS-activated RAW 264.7 cells and CLP-induced septic mice. Food Chem Toxicol. 2013;55:386–395.
  31. Lu S, Li D, Xi L, Calderone R. Interplay of interferon-gamma and macrophage polarization during Talaromyces marneffei infection. Microb Pathogen. 2019;134:103594.
  32. Zhong H, Yazdanbakhsh K. Hemolysis and immune regulation. Curr Opin Hematol. 2018;25(3):177–182.
  33. Hagmann M. A new way to keep immune cells in check. Science. 2000;288(5473):1945–1946.
  34. Oldenborg PA. Role of CD47 as a marker of self on red blood cells. Science. 2000;288(5473):2051–2054.
  35. Stachurska A, Dorman M, Korsak J, et al. Selected CD molecules and the phagocytosis of microvesicles released from erythrocytes ex vivo. Vox Sang. 2019;114(6):576–587.
  36. van Manen L, Peters AL, van der Sluijs PM, Nieuwland R, van Bruggen R, Juffermans NP. Clearance and phenotype of extracellular vesicles after red blood cell transfusion in a human endotoxemia model. Transfus Apher Sci. 2019;58(4):508–511.