Decreased cellular IL-23 but not IL-17 production in children with autism spectrum disorders

doi:10.1016/j.jneuroim.2009.09.005

Journal of Neuroimmunology

Volume 216, Issues 1–2, 30 November 2009, Pages 126-129

https://doi.org/10.1016/j.jneuroim.2009.09.005 Get rights and content

Abstract

A potential role for T_H17 cells has been suggested in a number of conditions including neurodevelopmental disorders such as autism spectrum disorders (ASD). In the current study, we investigated cellular release of IL-17 and IL-23 following an in-vitro immunological challenge of peripheral blood mononuclear cells (PBMC) from children with ASD compared to age-matched typically developing controls. Following stimulation, the concentration of IL-23, but not IL-17, was significantly reduced (p = 0.021) in ASD compared to controls. Decreased cellular IL-23 production in ASD warrants further research to determine its role on the generation and survival of T_H17 cells, a cell subset important in neuroinflammatory conditions that may include ASD.

Introduction

Autism spectrum disorders (ASD) are neurodevelopmental disorders characterized by severe impairments in social interaction and communication, and restricted, stereotyped interests (American Psychiatric Association, 1994). Symptoms of ASD generally manifest within the first three years of life and persist through adulthood in most cases. Evidence of immune dysregulation has been observed in some individuals with ASD, including increased levels of pro-inflammatory cytokines in brain tissues, CSF and plasma, and increased production of pro-inflammatory cytokines by peripheral blood mononuclear cell (PBMC) cultures when compared to typically developing controls (Ashwood et al., 2008, Ashwood and Wakefield, 2006, Enstrom et al., 2009a, Jyonouchi et al., 2001, Molloy et al., 2006, Vargas et al., 2005, Zimmerman et al., 2005).

Recently, T_H17 cells (Bettelli et al., 2007) were described as a subset of T cells unique for their production of interleukin (IL)-17, a cytokine demonstrated to be important for defense against extracellular bacteria (Raffatellu et al., 2008, Tato and O'Shea, 2006), fungi (Taylor et al., 2007), and specific parasites (Reece et al., 2008). In addition, these cells have been implicated in the pathology of a number of inflammatory conditions including multiple sclerosis (MS), rheumatoid arthritis, Crohn's disease, and psoriasis (Graber et al., 2008, Tzartos et al., 2008, Vaknin-Dembinsky et al., 2008). As a survival and proliferative factor for T_H17 cells, IL-23 promotes sustained production of IL-17 by T_H17 cells (Aggarwal et al., 2003). In animal models, anti-IL-23 therapy can ameliorate experimental autoimmune encephalomyelitis (EAE) (Chen et al., 2006), suggesting that dysregulation of T_H17 cells may contribute to disorders with suspected autoimmune/neuroinflammatory mechanisms, such as ASD.

Dysregulated immune responses in some individuals with ASD could lead to the aberrant production of IL-23 from immune cells. We recently reported that plasma IL-23 levels were decreased in ASD children (Enstrom et al., 2008). To help elucidate whether there exists a potential dysregulation of IL-23 and IL-17 production in ASD, we examined the effect of immune stimulation of PBMC isolated from children with ASD and typically developing controls in the same age range.

Section snippets

Materials and methods

Ninety-four subjects were recruited as part of the Childhood Autism Risks from Genetics and Environment (CHARGE) study (Hertz-Picciotto et al., 2006). For the experiments involving phytohemagglutinin (PHA) stimulation, 34 children with ASD (median age 3.83 (interquartile range 3.17–4.25), 29 males) and 26 typically developing (TD) controls (3.71 (3.00–4.50), 21 males) were included. Experiments involving phorbol myristate acetate (PMA) stimulation included 18 ASD children (4.25 (3.08–4.07), 14

Results

In the absence of PHA stimulation the levels of IL-23 and IL-17 were undetectable in cell culture supernatants from ASD or TD children. Following stimulation, IL-23 levels were significantly induced in cell culture supernatants from TD controls (median 43.69 pg/ml (interquartile range 13.31 pg/ml–78.34 pg/ml)) and were significantly higher than in PHA-stimulated cell culture supernatants from ASD children (9.09 pg/ml (4.50 pg/ml–24.52 pg/ml), p = 0.021, Fig. 1). Following stimulation IL-17 levels were

Discussion

The major finding of this study was that the production of IL-23 but not IL-17 following stimulation was significantly lower in children with ASD compared with TD controls. We previously reported that plasma levels of IL-23 but not IL-17 were decreased in children with ASD compared with TD controls (Enstrom et al., 2008). In addition, we found a negative correlation between stimulated IL-23 levels and ADOS scores of social interaction in ASD children. It is currently unclear how decreased IL-23

Acknowledgements

This study was funded by the NIEHS Children's Center grant (P01 ES011269), US EPA STAR Program grant (R833292 and R829388), NIEHS CHARGE study (R01ES015359), Cure Autism Now Foundation, Peter Emch Foundation, The Boler Company Foundation, HEDCO Foundation and a generous gift from the Johnson Family. We thank Isaac Pessah for his careful review and suggestions in the completion of this manuscript. We would like to thank the staff of both the UC Davis M.I.N.D. Institute and the CHARGE study for

References (32)

S. Aggarwal et al.
Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17
J. Biol. Chem.
(2003)
P. Ashwood et al.
Decreased transforming growth factor beta1 in autism: a potential link between immune dysregulation and impairment in clinical behavioral outcomes
J. Neuroimmunol.
(2008)
P. Ashwood et al.
Immune activation of peripheral blood and mucosal CD3+ lymphocyte cytokine profiles in children with autism and gastrointestinal symptoms
J. Neuroimmunol.
(2006)
E. Bettelli et al.
Th17: the third member of the effector T cell trilogy
Curr. Opin. Immunol.
(2007)
A.M. Enstrom et al.
Altered gene expression and function of peripheral blood natural killer cells in children with autism
Brain Behav. Immun.
(2009)
A. Enstrom et al.
Increased IgG4 levels in children with autism disorder
Brain Behav. Immun.
(2009)
J.J. Graber et al.
Interleukin-17 in transverse myelitis and multiple sclerosis
J. Neuroimmunol.
(2008)
H. Jyonouchi et al.
Proinflammatory and regulatory cytokine production associated with innate and adaptive immune responses in children with autism spectrum disorders and developmental regression
J. Neuroimmunol.
(2001)
C.A. Molloy et al.
Elevated cytokine levels in children with autism spectrum disorder
J. Neuroimmunol.
(2006)
J.S. Tzartos et al.
Interleukin-17 production in central nervous system-infiltrating T cells and glial cells is associated with active disease in multiple sclerosis
Am. J. Pathol.
(2008)

A. Vaknin-Dembinsky et al.

Increased IL-23 secretion and altered chemokine production by dendritic cells upon CD46 activation in patients with multiple sclerosis

J. Neuroimmunol.

(2008)

A.W. Zimmerman et al.

Cerebrospinal fluid and serum markers of inflammation in autism

Pediatr. Neurol.

(2005)

Y. Ziv et al.

Immune-based regulation of adult neurogenesis: implications for learning and memory

Brain Behav. Immun.

(2008)

American Psychiatric Association

Diagnostic and Statistical Manual of Mental Disorders

(1994)

Y. Chen et al.

Anti-IL-23 therapy inhibits multiple inflammatory pathways and ameliorates autoimmune encephalomyelitis

J. Clin. Invest.

(2006)

L.K. Curran et al.

Behaviors associated with fever in children with autism spectrum disorders

Pediatrics

(2007)

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Short communicationDecreased cellular IL-23 but not IL-17 production in children with autism spectrum disorders

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

J. Biol. Chem.

J. Neuroimmunol.

J. Neuroimmunol.

Curr. Opin. Immunol.

Brain Behav. Immun.

Brain Behav. Immun.

J. Neuroimmunol.

J. Neuroimmunol.

J. Neuroimmunol.

Am. J. Pathol.

J. Neuroimmunol.

Pediatr. Neurol.

Brain Behav. Immun.

Diagnostic and Statistical Manual of Mental Disorders

Anti-IL-23 therapy inhibits multiple inflammatory pathways and ameliorates autoimmune encephalomyelitis

J. Clin. Invest.

Behaviors associated with fever in children with autism spectrum disorders

Pediatrics

Short communication
Decreased cellular IL-23 but not IL-17 production in children with autism spectrum disorders