Elsevier

Immunobiology

Volume 216, Issues 1–2, January–February 2011, Pages 208-212
Immunobiology

High concentrations of anti-caspase-8 antibodies in Chilean patients with type 1 diabetes

https://doi.org/10.1016/j.imbio.2010.05.004Get rights and content

Abstract

Introduction

Deregulation of apoptosis across the Fas–FasL pathway is an increasingly relevant phenomenon in the pathogenic mechanisms associated with autoimmune diseases. Caspase-8 initiates the activation of the apoptotic process and interacts directly with Fas in the membrane of the T lymphocyte.

Objectives

To standardize an Elisa essay to measure the concentration of anti-caspase-8 antibodies in plasma of Type 1 Diabetes (T1D) patients and analyze their possible distribution and association with characteristics of the disease.

Methods and subjects

124 patients newly diagnosed with T1D and 132 controls: children and youngsters. ELISA test was standardized to detect anti-caspase-8 antibodies in plasma. It correlated the concentration of this antibody with classical markers of autoimmunity as anti-IA-2 and anti-GAD65, and the clinical characteristics at onset of diabetes mellitus. The statistical analysis was performed using logistic regression.

Results

Patients with T1D showed a higher concentration of anti-caspase-8 antibodies regarding the controls (87.5 ng/ml versus 24.3 ng/ml, p < 0.0001, values expressed as median). The proportion of patients with T1D and high concentrations of anti-caspase-8 (percentile 50–75) was significantly different from the control group (p < 0.0001). Anti-caspase-8 showed a strong association with positive anti-GAD65 (OR = 3.48, p < 0.035) and ketoacidosis (OR = 10.74, p < 0.0001) events, with glycemia and age at diagnosis as contributing variables.

Conclusion

This is the first report in the literature of levels of anti-caspase-8 antibodies in T1D through ELISA. The high concentration in patients with T1D, and its strong correlation with anti-GAD65 auto-antibodies, suggests a potential role of anti-caspase-8 auto-antibodies as surrogate marker autoimmunity in T1D patients.

Introduction

The incidence of type 1 diabetes (T1D) in Santiago, Chile has increased during the last years. These data are concordant with the observation that the incidence of T1D is increasing in Latin America and worldwide (Carrasco et al., 1996, Carrasco et al., 2006).

T1D is an autoimmune disease whereby antigen-specific T cells selectively destroy insulin-producing pancreatic β cells (Alizadeh and Koeleman, 2008, Mordes et al., 2004, Pirot et al., 2008, Anderson and Bluestone, 2005). The activated T cells first invade the islets, leading to insulitis. This is followed by destruction of the islets, mediated by a complex interaction between the activated lymphocytes, cytokines, and macrophages (Pirot et al., 2008, Gregersen and Behrens, 2006, Kawasaki et al., 2004). Apoptosis is a fundamental process involved in the destruction of β cells. More recently, β-cell apoptosis has also been shown to facilitate cross-presentation of islet antigens, an important initial step for priming or activation of β-cell-specific T cells required for disease onset (Pirot et al., 2008, Kawasaki et al., 2004). Dysfunction of apoptosis through the Fas–Fas ligand pathway is related to the onset of autoimmune diseases. Several studies have proposed that the blocking of apoptosis is involved in the pathogenesis of these autoimmune diseases (Pirot et al., 2008, Kawasaki et al., 2004, Pearl-Yafe et al., 2006).

Caspases are evolutionarily conserved cysteine-aspartyl specific proteases that play a key role in apoptosis (Reed, 2000). There are two main apoptotic pathways: the extrinsic or cell death receptor pathway involving the tumor necrosis factor super family of receptors and the intrinsic or mitochondrial pathway. Caspase-8 and caspase-9 are the upstream caspases involved in the extrinsic and intrinsic pathways, respectively. Caspase-3, -6, and -7 are effector caspases downstream of both pathways (Liadis et al., 2005). Different studies have reported the existence of impaired apoptosis in peripheral T cells in subjects affected by autoimmune diseases (Fujinami et al., 2006, Todaro et al., 2004). In subjects affected by Hashimoto thyroiditis and systemic sclerosis, a resistance to both Fas- and ceramide-induced apoptosis can occur in peripheral T cells, partially as a consequence of an impaired activity of caspase-8 (Stassi et al., 2001, Bossowski et al., 2007, Bossowski et al., 2008, Otsuki et al., 2006, Ueki et al., 2002, Yamaoka et al., 2008).

In the last years, caspase-8 has been extensively studied as the initiator caspase of the extrinsic apoptosis (Kang et al., 2004, Liadis et al., 2007, Beisner et al., 2005). However, a special attention has been observed related with the essential role in T-cell homeostasis and T-cell immunity (Salmena et al., 2003, Choi and Woo, 2010). These new functions of caspase-8 showed the possible role in cellular functions that are non-apoptotic as the activation of NF-κB, showing a dual role of this protein in cellular proliferation and apoptosis (Lemmers et al., 2007, Su et al., 2005, Maelfait and Beyaert, 2008, Tang et al., 2005). In T1D patients, several auto-antibodies against known and unknown antigens are present in the serum of recently diagnosed patients. In this study, we analyzed the serum concentration of anti-caspase-8 antibodies directed against caspase-8.

Section snippets

Subjects

The study was carried out in Santiago, Chile, with new cases of T1D diagnosed by means of standardized methods during 2006–2008. There were 124 new cases of T1D and 132 healthy unrelated control children. The control group represents volunteers who participated from 10 random schools from Santiago comparable for two Hispanic surnames and similar socioeconomic conditions. Information regarding the clinical characteristics of the debut, diagnosis data, allergies, familial history of diabetes and

Results

Patients with T1D have an age of diagnosis of 9.4 ± 4.7 years, glycemia of 456 ± 259, ketoacidosis of 56% and duration of breastfeeding of 5 ± 4.5 months. Clinical and demographic information is presented in Table 1. A majority of the subjects included in this study were below 15 years old in both groups. The patients were tested for anti-GAD65 and anti-IA-2 auto-antibodies, as well as anti-caspase-8 antibodies. We first compared the serum anti-caspase-8 antibody concentrations between T1D patients

Discussion

Type 1 diabetes (T1D) is a T cell-mediated autoimmune disease targeting pancreatic beta-cells. The process of β cell destruction, marked by the production of auto-antibodies, occurs over many years and ultimately results in metabolic abnormalities first manifested as impaired glucose tolerance (Knip and Siljander, 2008). The evaluation of isolated antibodies even in the population at risk of developing diabetes has a limited clinical value and for this reason, the screening of a battery of them

Competing interest

The authors declare that they have no competing interests.

Acknowledgements

This work was supported by FONDECYT Grant 1060790 (Prof. Francisco Pérez-Bravo). We thank all families participating in this investigation.

Authors’ contribution: FPB, AOA, ECD, ECP analyzed the data and drafted the manuscript. ECD and ECP provided subjects. AOA performed the serum analysis. LLM performed the statistical analysis. FPB advised on the technical protocol. All authors contributed to the conceptual design and analytical support. All authors read and approved the final manuscript.

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