Elsevier

Neurobiology of Disease

Volume 25, Issue 2, February 2007, Pages 239-251
Neurobiology of Disease

IgG entry and deposition are components of the neuroimmune response in Batten disease

https://doi.org/10.1016/j.nbd.2006.09.005Get rights and content

Abstract

Patients and a mouse model of Batten disease, the juvenile form of neuronal ceroid lipofuscinosis (JNCL), raise autoantibodies against GAD65 and other brain-directed antigens. Here we investigate the adaptive component of the neuroimmune response. Cln3−/− mice have autoantibodies to GAD65 in their cerebrospinal fluid and elevated levels of brain bound immunoglobulin G (IgG). IgG deposition was found within human JNCL autopsy material, a feature that became more evident with increased age in Cln3−/− mice. The lymphocyte infiltration present in human and murine JNCL occurred late in disease progression, and was not capable of central/intrathecal IgG production. In contrast, we found evidence for an early systemic immune dysregulation in Cln3−/− mice. In addition evidence for a size-selective breach in the blood–brain barrier integrity in these mice suggests that systemically produced autoantibodies can access the JNCL central nervous system and contribute to a progressive inflammatory response.

Introduction

Neuroinflammation is an important feature in many neurological conditions (Giovannoni and Baker, 2003), and is increasingly recognized as a pathogenic component in chronic neurodegenerative disorders such as Alzheimer's and Parkinson's disease (Perry, 2004). Inflammatory responses within the central nervous system (CNS) have also been reported in a growing number of childhood neurodegenerative conditions including GM1 and GM2 gangliosidoses (Wada et al., 2000, Jeyakumar et al., 2003, Yamaguchi et al., 2004), mucopolysaccaridoses I and IIIB (Ohmi et al., 2003), and the neuronal ceroid lipofuscinoses (NCLs) (Cooper, 2003, Pontikis et al., 2004). CNS inflammation can involve innate (glial activation) and/or adaptive (T cell and B cell activation) immunity, although whether each of these components contributes directly to the neurodegenerative process or whether it plays a neuroprotective role in these disorders is unclear (Nguyen et al., 2002, Schwartz and Kipnis, 2002, Streit, 2002). The neuronal ceroid lipofuscinoses (NCLs) are a group of at least eight genetically distinct lysosomal storage disorders (Hofmann and Peltonen, 2001, Cooper, 2003, Mitchison et al., 2004). These fatal autosomal recessive disorders have variable onset ranging from infancy to adulthood (Hofmann and Peltonen, 2001, Gardiner, 2002) with clinical signs including visual failure leading to blindness, an increased severity of untreatable seizures and neurocognitive decline followed by an inevitable premature death (Hofmann and Peltonen, 2001, Gardiner, 2002). Juvenile NCL (JNCL) or Batten disease is the most prevalent form of these disorders and is the result of mutations in the CLN3 gene, which encodes a transmembrane protein whose specific function remains unknown (The International Batten Disease Consortium, 1995).

The effect of CLN3 mutation on the CNS is profound, with widespread loss of cortical neurons (Braak and Goebel, 1978, Braak and Goebel, 1979), and regionally specific effects upon neuronal survival within the hippocampal formation (Tyynela et al., 2004). The generation of Cln3 null mutant mice (Cln3−/−) has provided a valuable model for studying the underlying neurodegenerative mechanisms which lead to these pathological changes (Mitchison et al., 1999). Neurologically, these Cln3−/− mice show only minor visual deficits by 12 months of age and do not display any overt spontaneous seizure activity, but die prematurely at approximately 18 months of age (Mitchison et al., 1999). Cln3−/− mice display the progressive neurodegenerative phenotype of JNCL exhibiting widespread accumulation of autofluorescent storage material from around the time of birth, CNS atrophy and the progressive loss of subpopulations of GABAergic interneurons that occur relatively late in disease progression, and an early loss of visual relay neurons in the thalamus at 6 months of age (Mitchison et al., 1999, Pontikis et al., 2004, Weimer et al., 2006).

There is also increasing evidence for a reactive component to JNCL pathogenesis, with regionally specific patterns of astrocytosis and microglial activation in human autopsy material (Tyynela et al., 2004). This response appears at low levels early in the course of disease and precedes neurodegeneration in Cln3−/− mice (Pontikis et al., 2004). Both Cln3−/− mice and individuals with JNCL also have circulating autoantibodies to glutamic acid decarboxylase (GAD65), reduced levels of brain GAD activity and presynaptic elevation of glutamate (Chattopadhyay et al., 2002a, Chattopadhyay et al., 2002b). Whether these GAD65 autoantibodies contribute to, or compound, the neuroinflammation reported in JNCL remains unclear, but this autoimmune response is unlikely to be confined solely to GAD65 (Pearce et al., 2004). Indeed, JNCL serum displays patterns of immunoreactivity against a variety of neuron subtypes suggesting that multiple brain-directed autoantibodies are raised in these individuals (Lim et al., 2006).

In this report we detail the adaptive component of the neuroimmune response in JNCL further, demonstrating the presence of anti-GAD65 antibodies within the cerebrospinal fluid (CSF) and an increased immunoglobulin G (IgG) deposition in the CNS of Cln3−/− mice and individuals with end stage JNCL. Investigation of the origin of these IgGs revealed evidence of an early systemic immune irregularity, with only limited lymphocyte infiltration into the JNCL CNS. Examination of how such systemically produced IgGs may access the CNS revealed evidence for size-selective breaches in blood–brain barrier (BBB) integrity in Cln3 null mutant mice. Taken together, these data suggest that an evolving neuroinflammatory response occurs in JNCL pathogenesis, which may influence ongoing neurodegeneration.

Section snippets

Mice

Cln3−/− mice inbred on a 129S6/SvEv background and control mice on the same strain background were used (Mitchison et al., 1999) and housed under standard non-sterile conditions until appropriately aged for each study. To avoid potential complications of hormonal influence, only male mice were used in all experiments. All perfusion and injection procedures (described below), were carried out in accordance with the NIH Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80-23)

Anti-GAD65 autoantibodies are present in the CSF

Circulating autoantibodies to glutamic acid decarboxylase (GAD65) are present in Cln3−/− mice and individuals with JNCL (Chattopadhyay et al., 2002a, Chattopadhyay et al., 2002b). To determine whether GAD65 autoantibodies are also present in the CSF of Cln3−/− mice we probed dot blots containing 10 ng recombinant human GAD (Kronus, USA) with 1:100 dilution of CSF from 3 month old Cln3−/− mice and age-matched controls (n = 3). Filters probed with CSF from Cln3−/− mice, followed by HRP-labeled

Discussion

This study represents the first detailed survey of the adaptive components of the autoimmune response in JNCL. Moreover, we have combined studies on human JNCL tissue and a mouse model of this disorder to evaluate more completely whether CLN3 deficiency leads to a modified autoimmune response. Our data reveal the presence of autoantibodies within CSF of Cln3−/− mice and IgG deposition within the JNCL CNS, starting early in disease progression. These IgGs do not appear to be produced within the

Acknowledgments

We would like to thank Andrew Serour and Timothy Curran for their excellent technical assistance; Drs. Jaana Tyynelä and Matti Haltia, University of Helsinki for providing human JNCL autopsy material; Dr. James Powers for useful discussions and Dr. Alison Barnwell for constructive comments on the manuscript. These studies were supported by National Institutes of Health grants NS044310 (D.A.P.), NS041930 European Commission 6th Framework Research Grant LSHM-CT-2003-503051 (J.D.C.), the WellChild

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