Elsevier

Journal of Hepatology

Volume 41, Issue 6, December 2004, Pages 1031-1037
Journal of Hepatology

Predominance of type 1 (Th1) cytokine production in the liver of patients with HCV-associated mixed cryoglobulinemia vasculitis

https://doi.org/10.1016/j.jhep.2004.08.011Get rights and content

Background/Aims

Patients with hepatitis C virus (HCV) mixed cryoglobulinemia (MC) vasculitis have a higher mortality rate and more frequent incidence of cirrhosis than their cryoglobulin-negative counterparts. To compare the cytokine profile of liver-infiltrating T cells in HCV-infected patients with or without MC vasculitis.

Methods

Hepatic biopsy specimens were obtained from HCV infected patients with and without MC vasculitis. Using intracellular staining and flow cytometry, we assessed the ability of freshly isolated liver T cells from these biopsies to produce IFN-γ, TNF-α, IL-2, IL-4, and IL-10 in response to stimulation with PMA and ionomycin.

Results

HCV-MC vasculitis patients compared to HCV-MC negative controls have an enhanced hepatic T cells production of Th1-type cytokines [i.e. TNF-α(30.3±13% vs. 15.5±5%, P=0.01), IL-2 (20.2±9% vs. 10±4%, P=0.01) and IFN-γ (22.2±11% vs. 9.4±4%, P=0.008)], whereas IL-10, a representative Th2-type cytokine, was significantly lower (7.2±4% vs. 17±7%, P=0.01).

Conclusions

T cell from the liver of HCV-MC vasculitis patients display a significantly augmented liver Th1 profile compared to MC-negative controls. This enhanced production of type-1 cytokines may account for a more severe course of liver disease.

Introduction

The syndrome of mixed cryoglobulinemia (MC) is characterized by the clinical triad of purpura, arthralgia, and asthenia. Cryoglobulins are composed of different immunoglobulins, of which the IgM rheumatoid factor (RF) component is monoclonal in type II MC and polyclonal in type III MC [1]. The clinical manifestations of MC result from an immune complex-type vasculitis (MC vasculitis) with deposition of immunoglobulins and complement in vasculitis lesions [2]. Although 60–90% of patients who are chronically infected with the hepatitis C virus (HCV) have cryoglobulin present in their serum, only a small proportion will develop a clinically symptomatic vasculitis [3]. Patients with HCV-MC vasculitis have a higher mortality rate and more frequent incidence of cirrhosis than their cryoglobulin-negative counterparts [4].

HCV has been recognized as the major etiologic factor of MC, possibly through B cell clonal proliferation stimulated by HCV antigens [5]. The liver histology of HCV-infected patients with MC shows a combination of portal and/or lobular inflammatory cell infiltration frequently associated with lymphoid nodules resembling secondary lymphoid organs [6]. Immunohistochemical characterization has demonstrated that they consist mainly of B cells surrounded by a T cell zone [7]. These follicle-like structures often display a well-formed germinal center and may act as true functional follicular structures [8]. The analysis of both B and T lymphocytes from the livers of HCV infected patients have provided particularly interesting data. The production of RF is associated with oligoclonal or monoclonal expansions of intrahepatic B cells [9]. The role of T cells at the site of lesions has been emphasized by Pham et al. [10]. Intrahepatic T cells have been shown to be clonally expanded. They express particular TCR Vβ gene products, suggesting an HCV-driven intrahepatic immune response.

An individual's Th1/2 differentiation is critical in influencing the liver disease progression and the course of HCV. Patients with severe liver disease, have enhanced expression of Type-1 cytokines [i.e. interferon (IFN)-γ, interleukin (IL)-2] [11]. In contrast, the prevalent cytokine pattern of circulating virus-specific T cells from patients who recover spontaneously from acute hepatitis is Th1-like and a prevalent Th2 pattern of cytokine production is associated with viral persistence and chronic evolution [12], [13]. Augmented Th1 cytokine production is also seen in patients that respond favorably to IFN-α therapy [14].

It has been suggested that HCV-MC patients are at higher risk of developing cirrhosis than HCV-cryoglobulin negative patients [3], [4], [15]. Recent data indicate that peripheral blood monocytes from HCV-related MC vasculitis patients produce higher level of Th1 polarizing cytokines than their cryoglobulin-negative counterparts [16]. However, to date, there is no available information on the cytokine profile of liver-infiltrating T cells in MC patients.

Since the liver is the primary site of HCV infection, we investigated the cytokine profile of liver T cells from HCV-infected patients with and without MC vasculitis. Our data indicate that a type-1 cytokine pattern clearly predominates in hepatic T cells from HCV-MC vasculitis patients.

Section snippets

Patients

The study received approval from the institutional ethics committee and informed consent was obtained from all patients. Nineteen HCV-infected patients (median age 52.6 years) entered the study, seven of whom had symptomatic MC. All had histologically proven chronically active liver disease, and positive HCV antibodies and RNA. They were negative for hepatitis B surface antigen and human immunodeficiency virus antibodies. MC-positive patients had serum MC >0.05 g/l, on at least two occasions.

Patients

The main characteristics of the 19 HCV-infected patients are summarized in Table 1. Baseline characteristics of the 7 patients with MC vasculitis were as follows: sex ratio male to female 0.75, median age 53.8 (39–71) years, neuropathy present in 85% (n=6) and nephropathy in 14% (n=1). All patients had a type II, IgM kappa MC with a mean cryoglobulin level of 1.8 (0.25–10.3) g/l. The mean duration of HCV infection was 16 (14–37) years and 19 (10–52) for MC patients and controls, respectively.

Discussion

Many CD3+ T cells infiltrate the liver of patients with chronic HCV infection. Besides minor subsets such as NK-T cells, these CD3+ liver-infiltrating cells are mostly conventional CD4+ and CD8+ T lymphocytes which have the potential to secrete type-1 or type-2 cytokines [23]. To analyze the type of effector lymphocytes present in the liver of HCV-infected patients, we activated freshly isolated hepatic T cells and assayed their production of TNF-α, IFN-γ, IL-2, IL-4, and IL-10 at the

Acknowledgements

Supported by Agence nationale de Recherche contre le SIDA, Fondation pour la recherche médicale, Faculté de Médecine Pitié-Salpêtrière and Assistance Publique-Hôpitaux de Paris.

References (38)

  • J.C. Brouet et al.

    Biologic and clinical significance of cryoglobulins. A report of 86 cases

    Am J Med

    (1974)
  • M. Casato et al.

    Predictors of long-term response to high-dose interferon therapy in type II cryoglobulinemia associated with hepatitis C virus infection

    Blood

    (1997)
  • P. Cacoub et al.

    Extrahepatic manifestations of chronic hepatitis C. MULTIVIRC Group

    Arthritis Rheum

    (1999)
  • F. Dammacco et al.

    Hepatitis C virus infection and mixed cryoglobulinemia: a striking association

    Int J Clin Lab Res

    (1993)
  • J. Murakami et al.

    Functional B-cell response in intrahepatic lymphoid follicles in chronic hepatitis C

    Hepatology

    (1999)
  • J.F. Mosnier et al.

    The intraportal lymphoid nodule and its environment in chronic active hepatitis C: an immunohistochemical study

    Hepatology

    (1993)
  • D. Sansonno et al.

    Clonal analysis of intrahepatic B cells from HCV-infected patients with and without mixed cryoglobulinemia

    J Immunol

    (1998)
  • C. Ferrari et al.

    Antiviral cell-mediated immune responses during hepatitis B and hepatitis C virus infections

    Recent Results Cancer Res

    (1998)
  • C. Prezzi et al.

    Virus-specific CD8(+) T cells with type 1 or type 2 cytokine profile are related to different disease activity in chronic hepatitis C virus infection

    Eur J Immunol

    (2001)
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