Inflammatory gene expression in Coxsackievirus B-4-infected human islets of Langerhans

https://doi.org/10.1016/j.bbrc.2005.03.016Get rights and content

Abstract

The event that triggers the autoimmune destruction of insulin-producing β-cells in type 1 diabetes mellitus (T1DM) is still unknown. Enterovirus, especially Coxsackievirus, infections have long been associated with this disease. Cytokines and chemokines induced by an enterovirus infection may act to trigger the autoimmune reactions that produce T1DM. Gene expression was examined in isolated human islets infected with a Coxsackievirus-B4 (CBV-4) strain causing lytic infection (V89-4557) and in islets infected with a CBV-4 strain establishing persistent infection (VD2921). Microarray analysis indicated that infection with the CBV-4 strains resulted in specific induction of a number of inflammatory genes, including IL-1β, IL-6, IL-8, MCP-1, and RANTES. Importantly, the inflammatory genes induced by the CBV-4 infections differed in the two strains, with more cytokines being induced by the non-lytic CBV-4 strain than by the lytic strain. These cytokines and chemokines have the potential to rapidly induce inflammatory reactions when expressed in vivo and could contribute to the autoimmune reactions associated with the development of T1DM.

Section snippets

Materials and methods

Isolation and culture of pancreatic islets. Using a protocol approved by the Local Ethical Committee, human islets of Langerhans were isolated from brain-dead donors free of known pancreatic disease as previously described [19]. The islet preparation was of good quality and was available for research only because of too low islet mass yield after isolation.

Islets were cultured in CMRL-1066 (Gibco-BRL, Invitrogen) supplemented with 10 mM nicotinamide (Sigma Chemicals), 10 mM Hepes buffer

Islet quality

The purity of the isolated islets was 80–90%. We evaluated islet function using a dynamic perfusion system (Fig. 1), in which insulin release was increased when the concentration of glucose in the perfusate was raised.

TCID50 titrations and degree of CPE

Both strains replicated well in the human islet cells (Fig. 2). Islets infected with the VD2921 strain showed no CPE while islets infected with the lytic strain V89-4557 exhibited CPE at the 1+ to 2+ level on Day 3 post-infection.

Microarray analysis

Interleukin (IL)-1β, IL-6, and IL-8 were

Discussion

In the present study, we have used two different strains of CBV-4 to investigate whether CBV-4-infected islets express inflammation-related genes whose expression could contribute to the autoimmune process that destroys the islets and finally leads to T1DM. We also asked whether the gene expression of these genes differed according to viral phenotype (lytic vs. non-lytic). Islets were either infected with V89-4557, a CBV-4 strain that has been shown to cause lytic infection in human islets in

Acknowledgments

The authors thank Susanne Gabrielsson for expert support in microarray analysis and The Juvenile Diabetes Research Foundation, The Swedish Diabetes Association, The Family Ernfors Fund, The Gillbergska Foundation, Swedish Animal Welfare Agency, Barndiabetesförbundet, the Ronald MacDonald Fund, and the Swedish Medical Research Council (K98-12XC-12445-02B) for financial support.

References (37)

  • M. Roivainen et al.

    Mechanisms of coxsackievirus-induced damage to human pancreatic beta-cells

    J. Clin. Endocrinol. Metab.

    (2000)
  • P. Ylipaasto et al.

    Enterovirus infection in human pancreatic islet cells, islet tropism in vivo and receptor involvement in cultured islet beta cells

    Diabetologia

    (2004)
  • J.W. Yoon et al.

    Isolation of a virus from the pancreas of a child with diabetic ketoacidosis

    N. Engl. J. Med.

    (1979)
  • H. Hyoty et al.

    A prospective study of the role of coxsackie B and other enterovirus infections in the pathogenesis of IDDM. Childhood Diabetes in Finland (DiMe) Study Group

    Diabetes

    (1995)
  • H. Hyoty et al.

    The role of viruses in human diabetes

    Diabetologia

    (2002)
  • G. Frisk et al.

    High frequency of Coxsackie-B-virus-specific IgM in children developing type I diabetes during a period of high diabetes morbidity

    J. Med. Virol.

    (1985)
  • G. Frisk et al.

    Coxsackie B virus IgM in children at onset of type 1 (insulin-dependent) diabetes mellitus: evidence for IgM induction by a recent or current infection

    Diabetologia

    (1992)
  • H. Yin et al.

    Enterovirus RNA is found in peripheral blood mononuclear cells in a majority of type 1 diabetic children at onset

    Diabetes

    (2002)
  • Cited by (0)

    View full text