Skip to main content

Advertisement

SpringerLink
Log in

HCV-Related Proteins Activate Kupffer Cells Isolated from Human Liver Tissues

  • Original Article
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Purpose

It was reported from this laboratory that Kupffer cells (KCs) were activated in patients infected with HCV. Since dendritic cells, monocytes, and macrophages were activated by stimulation with HCV-related proteins, the specific aim of this study was to investigate the role of HCV-related proteins in activation of KCs, the signal pathway of activation of KCs mediated by Toll-like receptor (TLR) 4, and the influence of HCV infection on function of KCs.

Methods

Kupffer cells isolated from non-cancerous surgical specimen were co-cultured with HCV-related proteins (Core, NS3, NS4, and NS5), and production of cytokines (TNF-α, IL-1β, and IL-10) and hydrogen peroxide were assessed. Furthermore, effects of neutralization antibodies against the TLR2, TLR3, or TLR4, and cytochalasin B on the production TNF-α by KCs were investigated.

Results

Kupffer cells produced markedly a proinflammatory cytokine TNF-α by stimulation with all HCV-related proteins studied, and values were as same as production by KCs stimulated with LPS. Importantly, this production in the case of NS3 was significantly blunted by about 60% by neutralization antibodies against the TLR4, but not cytochalasin B. Production of TNF-α by isolated KCs stimulated with LPS was significantly greater in the HCV-infected livers than the HCV/HBV-negative livers.

Conclusions

These results indicated that HCV-related proteins may cause prolonged activation of KCs in the HCV-infected liver, leading to accumulation of inflammatory cytokines that contribute to DNA damage and carcinogenesis. Furthermore, function of KCs was difference between patients infected with and without HCV infection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

DMSO:

Dimethyl sulfoxide

ELISA:

Enzyme-linked immunosorbent assay

HCC:

Hepatocellular carcinoma

HCV:

Hepatitis C virus

ICAM-1:

Intercellular adhesion molecule-1

IL:

Interleukin

KC:

Kupffer cell

LPS:

Lipopolysaccharide

MIP:

Macrophage inflammatory protein

NS3:

Nonstructural 3 protein

NS4:

Nonstructural 4 protein

NS5:

Nonstructural 5 protein

t-BOOH:

t-Butyl hydroperoxide

TLR:

Toll-like receptor

TNF:

Tumor necrosis factor

References

  1. Kiyosawa K, Tanaka E. Characteristics of hepatocellular carcinoma in Japan. Oncology. 2002;62(1):5–7.

    Article  PubMed  Google Scholar 

  2. Yoshizawa H. Hepatocellular carcinoma associated with hepatitis C Virus infection in Japan: projection to other countries in the foreseeable future. Oncology. 2002;62(Suppl 1):8–17.

    Article  PubMed  Google Scholar 

  3. Tanaka Y, Hanada K, Mizokami M, et al. Inaugural article: a comparison of the molecular clock of hepatitis C virus in the United States and Japan predicts that hepatocellular carcinoma incidence in the United States will increase over the next two decades. Proc Natl Acad Sci USA. 2002;99:15584–15589.

    Article  PubMed  CAS  Google Scholar 

  4. McGuinness PH, Painter D, Davies S, McCaughan GW. Increases in intrahepatic CD68 positive cells, MAC387 positive cells, and proinflammatory cytokines (particularly interleukin 18) in chronic hepatitis C infection. Gut. 2000;46:260–269.

    Article  PubMed  CAS  Google Scholar 

  5. Maki A, Kono H, Gupta M, et al. Predictive power of biomarkers of oxidative stress and inflammation in patients with hepatitis C virus-associated hepatocellular carcinoma. Ann Surg Oncol. 2007;14:1182–1190.

    Article  PubMed  Google Scholar 

  6. Matsumoto K, Satoh Y, Sugo H, et al. Immunohistochemical study of the relationship between 8-hydroxy-2′-deoxyguanosine levels in noncancerous region and postoperative recurrence of hepatocellular carcinoma in remnant liver. Hepatol Res. 2003;25:435–441.

    Article  PubMed  CAS  Google Scholar 

  7. Burgio VL, Ballardini G, Artini M, Caratozzolo M, Bianchi FB, Levrero M. Expression of co-stimulatory molecules by Kupffer cells in chronic hepatitis of hepatitis C virus stiology. Hepatology. 1998;27:1600–1606.

    Article  PubMed  CAS  Google Scholar 

  8. Wheeler MD, Kono H, Yin M, et al. The role of Kupffer cell oxidant production in early ethanol-induced liver disease. Free Radic Biol Med. 2001;31:1544–1549.

    Article  PubMed  CAS  Google Scholar 

  9. Rusyn I, Kadiiska MB, Dikalova A, et al. Phthalates rapidly increase production of reactive oxygen species in vivo: role of Kupffer cells. Mol Pharmacol. 2001;59:744–750.

    PubMed  CAS  Google Scholar 

  10. Dolganiuc A, Oak S, Kodys K, et al. Hepatitis C core and nonstructural 3 proteins trigger toll-like receptor 2-mediated pathways and inflammatory activation. Gastroenterology. 2004;127:1513–1524.

    Article  PubMed  CAS  Google Scholar 

  11. Dolganiuc A, Kodys K, Kopasz A, et al. Hepatitis C virus core and nonstructural protein 3 proteins induce pro-and anti-inflammatory cytokines and inhibit dendritic cell differentiation. J Immunol. 2003;170:5615–5624.

    PubMed  CAS  Google Scholar 

  12. Chang S, Dolganiuc A, Szabo G. Toll-like receptors 1 and 6 are involved in TLR2-mediated macrophage activation by hepatitis C virus core and NS3 proteins. J Leukoc Biol. 2007;82:479–487.

    Article  PubMed  CAS  Google Scholar 

  13. Heuff G, Meyer S, Beelen RHJ. Isolation of rat and human Kupffer cells by a modified enzymatic assay. J Immunol Meth. 1994;174:61–65.

    Article  CAS  Google Scholar 

  14. Decker K. Biologically active products of stimulated liver macrophages (Kupffer cells). Eur J Biochem. 1990;192:245–261. Review.

    Article  PubMed  CAS  Google Scholar 

  15. Pileri P, Uematsu Y, Campagnoli S, et al. Binding of hepatitis C virus to CD81. Science. 1998;282:938–941.

    Article  PubMed  CAS  Google Scholar 

  16. Zeisel MB, Koutsoudakis G, Schnober EK, et al. Scavenger receptor class B type I is a key host factor for hepatitis C virus infection required for an entry step closely linked to CD81. Hepatology. 2007;46:1722–1731.

    Article  PubMed  CAS  Google Scholar 

  17. Muriel P, Alba N, Perez-Alvarez VM, Shibayama M, Tsutsumi VK. Kupffer cells inhibition prevents hepatic lipid peroxidation and damage induced by carbon tetrachloride. Comp Biochem Physiol C Toxicol Pharmacol. 2001;130:219–226.

    Article  PubMed  CAS  Google Scholar 

  18. Niemela O, Parkkila S, Bradford B, Iimuro Y, Pasanen M, Thurman RG. Effect of Kupffer cell inactivation on ethanol-induced protein adducts in the liver. Free Radic Biol Med. 2002;33:350–355.

    Article  PubMed  CAS  Google Scholar 

  19. Michael SL, Pumford NR, Mayeux PR, Niesman MR, Hinson JA. Pretreatment of mice with macrophage inactivators decreases acetaminophen hepatotoxicity and the formation of reactive oxygen and nitrogen species. Hepatology. 1999;30:186–195.

    Article  PubMed  CAS  Google Scholar 

  20. Ju C, Reilly TP, Bourdi M, et al. Protective role of Kupffer cells in acetaminophen-induced hepatic injury in mice. Chem Res Toxicol. 2002;15:1504–1513.

    Article  PubMed  CAS  Google Scholar 

  21. Schwabe RF, Brenner DA. Mechanisms of liver injury. I. TNF-alpha-induced liver injury: role of IKK, JNK, and ROS pathways. Am J Physiol Gastrointest Liver Physiol. 2006;290:583–589.

    Article  Google Scholar 

  22. Kamata H, Honda S, Maeda S, Chang L, Hirata H, Karin M. Reactive oxygen species promote TNFα-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell. 2005;120:649–661.

    Article  PubMed  CAS  Google Scholar 

  23. Pham CG, Bubici C, Zazzeroni F, et al. Ferritin heavy chain upregulation by NF-kappaB inhibits TNFalpha-induced apoptosis by suppressing reactive oxygen species. Cell. 2004;119:529.

    Article  PubMed  CAS  Google Scholar 

  24. Asakawa M, Kono H, Amemiya H, et al. Role of interleukin-18 and its receptor in hepatocellular carcinoma associated with hepatitis C virus infection. Int J Cancer. 2006;118:564–570.

    Article  PubMed  CAS  Google Scholar 

  25. Zhang H-Y, Nanji AA, Luk JM, et al. Macrophage migration inhibitory factor expression correlates with inflammatory changes in human chronic hepatitis B infection. Liver Int. 2005;25:571–579.

    Article  PubMed  CAS  Google Scholar 

  26. Kallinowski B, Haseroth K, Marinos G, et al. Induction of tumour necrosis factor (TNF) receptor type p55 and p75 in patients with chronic hepatitis C virus (HCV) infection. Clin Exp Immunol. 1998;111:269–277.

    Article  PubMed  CAS  Google Scholar 

  27. Farinati F, Cardin R, Bortolami M, Guido M, Rugge M. Oxidative damage, pro-inflammatory cytokines, TGF-alpha and c-myc in chronic HCV-related hepatitis and cirrhosis. World J Gastroenterol. 2006;12:2065–2069.

    PubMed  CAS  Google Scholar 

  28. Tomita M, Yamamoto K, Kobashi H, Ohmoto M, Tsuji T. Immunohistochemical phenotyping of liver macrophages in normal and diseased human liver. Hepatology. 1994;20:317–325.

    Article  PubMed  CAS  Google Scholar 

  29. Dolganiuc A, Garcia C, Kodys K, Szabo G. Distinct Toll-like receptor expression in monocytes and T-cells in chronic HCV infection. World J Gastroenterol. 2006;12:1198–1204.

    PubMed  CAS  Google Scholar 

  30. Lin RS, Lee FY, Lee SD, et al. Endotoxemia in patients with chronic liver diseases: relationship to severity of liver diseases, presence of esophageal varices, and hyperdynamic circulation. J Hepatol. 1995;22:165–172.

    Article  PubMed  CAS  Google Scholar 

  31. Plachouras D, Stamatakos M, Baziaka F, et al. Portal and systemic endotoxemia in abdominal operations: the significance of acute abdomen. J Surg Res. 2006;134:133–137.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. First Department of Surgery, Faculty of Medicine, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi, 409-3898, Japan

    Naohiro Hosomura, Hiroshi Kono, Masato Tsuchiya, Kenichi Ishii, Masahito Ogiku, Masanori Matsuda & Hideki Fujii

Authors
  1. Naohiro Hosomura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hiroshi Kono
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masato Tsuchiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenichi Ishii
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masahito Ogiku
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masanori Matsuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hideki Fujii
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroshi Kono.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hosomura, N., Kono, H., Tsuchiya, M. et al. HCV-Related Proteins Activate Kupffer Cells Isolated from Human Liver Tissues. Dig Dis Sci 56, 1057–1064 (2011). https://doi.org/10.1007/s10620-010-1395-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10620-010-1395-y

Keywords

Search

Navigation