Table of Contents Author Guidelines Submit a Manuscript
Mediators of Inflammation
Volume 2013, Article ID 609602, 8 pages
http://dx.doi.org/10.1155/2013/609602
Research Article

Gestational Exposure to a Viral Mimetic Poly(I:C) Results in Long-Lasting Changes in Mitochondrial Function by Leucocytes in the Adult Offspring

1Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
2Medical Investigations of Neurodevelopmental Disorders (MIND) Institute, University of California, Davis, CA 95616, USA
3Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis, CA 95616, USA

Received 15 July 2013; Accepted 16 August 2013

Academic Editor: Giuseppe Valacchi

Copyright © 2013 Cecilia Giulivi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Maternal immune activation (MIA) is a potential risk factor for autism spectrum disorder (ASD) and schizophrenia (SZ). In rodents, MIA results in changes in cytokine profiles and abnormal behaviors in the offspring that model these neuropsychiatric conditions. Given the central role that mitochondria have in immunity and other metabolic pathways, we hypothesized that MIA will result in a fetal imprinting that leads to postnatal deficits in the bioenergetics of immune cells. To this end, splenocytes from adult offspring exposed gestationally to the viral mimic poly(I:C) were evaluated for mitochondrial outcomes. A significant decrease in mitochondrial ATP production was observed in poly(I:C)-treated mice (45% of controls) mainly attributed to a lower complex I activity. No differences were observed between the two groups in the coupling of electron transport to ATP synthesis, or the oxygen uptake under uncoupling conditions. Concanavalin A- (ConA-) stimulated splenocytes from poly(I:C) animals showed no statistically significant changes in cytokine levels compared to controls. The present study reports for the first time that MIA activation by poly(I:C) at early gestation, which can lead to behavioral impairments in the offspring similar to SZ and ASD, leads to long-lasting effects in the bioenergetics of splenocytes of adult offspring.

1. Introduction

The most recent estimates indicate that the prevalence of autism spectrum disorders (ASD) in the United States has raised to 1 in 54 boys and 1 in 252 girls [1]. Although increased awareness and changes in diagnostic criteria have been proposed as the major contributors to this increased prevalence [2], as of today, the etiopathology of disorders like ASD and schizophrenia (SZ) remains largely unknown.

Several studies have suggested that impaired mitochondrial function and altered energy metabolism in individuals with ASD may contribute to their social and cognitive deficits [35], and recent reports indicate the presence of mitochondrial dysfunction (MD) in brain, skeletal muscle, and peripheral blood mononuclear cells (PBMC) from children with ASD. The MD in ASD is generally characterized by lower complex I activity accompanied, in a subset of cases, by deficits in other complexes [68].

Beside their critical role in a number of pathways, spanning from ATP production (via oxidative phosphorylation), one-carbon metabolism regulation, heme biosynthesis, fatty acid catabolism, and branched chain amino acid metabolism [9], mitochondria may also impact the immune response and vice versa [1012]. For example, human neutrophil mitochondria are involved in several functions such as chemotaxis, respiratory burst activity, maintenance of cell shape, and apoptosis [1317]. Furthermore, neutrophil phagocytosis may involve the incorporation of some mitochondrial proteins into the phagosome [18]. In addition, mitochondria can be involved in the immune response by providing part of the metabolic pathway for Gln, in a process named “glutaminolysis” [19, 20]. Interestingly, Gln is implicated in the expression of  NADPH oxidase components, cytokine production in lymphocytes, and macrophage, and as a provider for substrates required for nucleic acid synthesis [2125]. Taken together, these lines of evidence unveil a link between mitochondria and immune response [1012]. Indeed, deficits in bioenergetics have been reported in lymphocytes from children with ASD enrolled in the case-control population-based Childhood Autism Risk Genetics and Environment (CHARGE) Study [26, 27]. Children with ASD in this study display a number of immune dysfunctions including abnormalities in monocytes, T cells and NK cell responses [28]. These observations suggest the presence of a genetic background that results in a distinct immune profile in responses to a variety of triggers, among them psychological stressors, exposure to chemical triggers, and infectious agents [29, 30].

Considering that (i) mitochondria are inherited maternally via oocyte, (ii) maternal diet or immune activation during pregnancy has an impact on fetal metabolic and immune programming [3133], and (iii) offspring born to pregnant mice injected with poly(inosinic:polycytidylic acid) (poly(I:C)), a synthetic double-stranded RNA that mimics viral infection via activation of Toll-like receptor-3 (TLR3), at embryonic day 12.5 (E12.5), display core behavioral symptoms of ASD [34, 35] and SZ [34], it is hypothesized that prenatal exposure of mothers to an immunogenic response, that is, poly(I:C) elicits changes in mitochondrial function in splenocytes from progeny lasting into adulthood. Exposure to TLR ligands can lead to maternal hypertension, vascular dysfunction, and proteinuria in pregnant animals but not in nonpregnant animals [3638] suggesting the occurrence of a differential immune response/pathway during pregnancy. Differences are also evident between pregnant individuals with human placentas and patients with preeclampsia showing greater expression of TLR3, along with TLR2, TLR4, and TLR9, compared to nonpreeclampsia mothers [39, 40]. These data suggest that TLR signaling may be involved in placental deficiencies/abnormalities that may provide a framework for altered fetal programming. Of note, trophoblastic inclusions, which are also observed in preeclampsia and other placental defects, were reported to be increased in placenta from mothers of children with ASD compared to controls [41]. Furthermore, maternal exposure to various pathogens, including viruses, significantly increases the risk for ASD and SZ [4248]. Considering that maternal exposure to various pathogens is associated with ASD and SZ, the critical link between prenatal maternal infection and postnatal brain and behavioral pathology seem to be the maternal immune response, including cytokine production [47, 4954], which may contribute to the fetal imprinting of the neuroimmune response and, possibly, mitochondria-mediated metabolic responses. Although it is already known that upon poly(I:C) injection, the induction of maternal cytokines alters the expression of several cytokines in the fetal brain (with only IL-1β remaining elevated at 24 h [53] with only a few changes during adulthood), it is unknown if maternal immune activation (MIA) also causes chronic changes in the bioenergetics of immune cells (such as splenocytes) of adult offspring.

In this study, we sought to determine whether MIA in pregnant dams alters mitochondrial function in splenocytes from affected offspring. To test this, dams were exposed to poly(I:C) on gestational day 12.5 to induce MIA. This stage of gestation correlates with the late first trimester in humans [55], coincidental with the time that infections are most closely linked to increased incidence of ASD and SZ [47, 48]. The present study reports for the first time that MIA activation by poly(I:C) at early gestation, which can lead to impairments in multiple psychological domains, is associated with mitochondrial changes in immune the cells of adult offspring.

2. Materials and Methods

2.1. Animals

Male and female C57BL/6J (Jackson Laboratory, Sacramento, CA, USA) mice were bred and maintained by the Center for Laboratory Animal Research, at University of California, Davis, and maintained at ambient room temperature on a 12 h light/dark cycle (lights on at 06:00 h). Food and water were provided ad libitum. All procedures were performed with approval by the Institutional Animal Care and Use Committee, University of  California, Davis, and in accordance with the guidelines provided by the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

2.2. Treatment and Behavioral Assessment

Mice were mated overnight and females were checked daily for the presence of seminal plugs, noted on gestational day 0.5 (G0.5). On G12.5, pregnant female mice were weighed and injected with a single dose (20 mg/kg; i.p.) of poly(I:C) (Sigma Aldrich, St Louis, MO, USA) or saline vehicle (SHAM) as previously described [35]. Each dam was returned to its cage and left undisturbed until the birth of its litter. All mice pups remained with the mother until weaning on postnatal day 21, at which time mice were group-housed 3-4 per cage with same-sex littermates. Mice born from poly(I:C)-treated dams exhibited autism-like behavioral deficits including reduced social approach, increased ultrasonic vocalizations, and repetitive marble burying behaviors [35].

2.3. Splenocyte Isolation

One week following behavioral testing, 12 wk old mice were sacrificed by cervical dislocation and spleens were collected for tissue processing. While spleen is constituted by a variety of cells relevant to the immune response (including T- and B-lymphocytes, dendritic cells, and macrophages), it has recently been shown that spleens from the offspring of MIA mice elicited by poly(I:C) provide a more homogeneous preparation enriched in granulocytes compared to the preparation obtained from whole blood [56]. Briefly, spleens were homogenized into single cell suspensions by gently pushing them through a 100 μm nylon mesh filter (Fisher Sci) into PBS at 4°C. Cells were then pelleted, and RBCs were lysed using ACK lysis buffer according to the manufacturer’s instructions (Gibco). Cell suspensions were kept on ice until analyzed for mitochondrial activity. Cell viability was determined by trypan blue staining and found to be about 90%.

2.4. Mitochondrial Activities

The oxygen uptake of  intact cell suspensions (106 cells/mL) obtained as described above was measured by using a Clark-type O2 electrode from Hansatech (King’s Lynn, UK) at 22°C. Cells were incubated in the presence of 5 mM glucose in calcium and magnesium-supplemented HBSS buffer without phenol red at 20–22°C. NADH, succinate, and cytochrome oxidase activities were evaluated under phosphorylating conditions as described before [6, 30]. To this end, cells were permeabilized with a controlled treatment with digitonin [57] by adding 60 μg/mL 2x recrystallized digitonin for 2 min. The solubilization was stopped by the addition of 1 mg/mL BSA. Oxygen consumption rates were evaluated in the presence of 1 mM ADP plus 1 mM malate-10 mM glutamate followed by the addition of 5 μM rotenone; 10 mM succinate followed by the addition of 3.6 μM antimycin A, and 10 mM ascorbate and 0.2 mM N,N,N,N-tetramethyl-p-phenylenediamine followed by the addition of 1 mM KCN.

2.5. Statistical Analyses

All mitochondrial experiments were run in triplicates. Mitochondrial data were expressed as mean ± standard error. Student’s two-tailed -test was used to evaluate the differences between offspring of poly(I:C)-treated and SHAM-treated dams.

3. Results and Discussion

3.1. Deficits in Complex I in Splenocytes from Mice Gestationally Exposed to Poly(I:C)

Splenocytes from adult mice born to either SHAM- or poly(I:C)-treated dams were isolated for mitochondrial function testing. Given that most of the oxygen uptake by cells is linked to ATP production via oxidative phosphorylation, this parameter was evaluated in intact cells in the presence of glucose (Figure 1(a)). The rate of oxygen uptake by intact cells from SHAM-treated dams was 0.22 ± 0.03 nmol oxygen × (min × 106 cells)−1. Under the same conditions, this rate was decreased by 36% in poly(I:C)-treated dams (Figure 1(a)). Addition of oligomycin, an inhibitor of ATPase, was used to stop the fraction of oxygen utilized to synthesize ATP via mitochondria. In both groups, more than 90% of the total oxygen uptake was inhibited by oligomycin, supporting the previous assumption that most—if not all—oxygen uptake by these cells was derived from oxidative phosphorylation. The oxygen uptake resistant to oligomycin, considered somewhat equivalent to State 4 (nonphosphorylating mitochondria), was not different between groups. This result indicated that the proton leak across the inner mitochondrial membrane was similar in both groups, suggesting no major mitochondrial membrane damage by either treatment. Addition of FCCP, an uncoupler of electron transport and ATP synthesis, increased significantly the basal oxygen uptake to a similar extent in both groups (2.5- to 3-fold) with no changes between treatments, suggesting that the maximum respiratory capacity was similar in both groups.

fig1
Figure 1: Mitochondrial outcomes in splenocytes. Splenocytes from offspring (10–12 weeks old) poly(I:C)- or saline- treated dams at 12.5 gestational day were isolated, and mitochondrial outcomes were tested as described in detail in Section 2. (a) Oxygen uptake of intact cells was determined using a Clark-type electrode in Hanks balanced salt solution (HBSS) supplemented with 6 mM glucose as substrate. Basal (only glucose), oligomycin (0.2 μM), and FCCP (20 μM) were added, and the initial rates of oxygen uptake were calculated and normalized by million cells. (b) Permeabilized cells were tested for their capability to consume oxygen coupled to ATP production by supplementing the media with malate-glutamate (or NADH oxidase comprised by complexes I–V), succinate with rotenone (or succinate oxidase comprised by complex II–V), and TMP-ascorbate for cytochrome c oxidase activity (or complex IV). (c) ratio of rates of oxygen consumption in the presence of various substrates derived from (b).

Coupling between oxygen uptake and ATP production was evaluated by the respiratory control ratio in intact cells (RCR). Mitochondria from either treatment showed a significant coupling with glucose as a substrate (with malate-glutamate, RCR = 3.5 ± 0.4 and 2.7 ± 0.3; with succinate, RCR = 4 ± 1 and 6.2 ± 0.5, for saline and poly(I:C), resp.) with no statistical differences between treatment groups. This result indicated that mitochondria were highly coupled and provided a means of support for their integrity during the testing process.

Phosphorylating mitochondria from splenocytes of SHAM animals in the presence of an NAD-linked substrate (such as malate) consumed oxygen at a rate of 0.31 ± 0.04 nmol oxygen × (min × 106 cells)−1. Phosphorylating mitochondria from offspring of poly(I:C)-treated dams showed a significant decrease in oxygen consumption (by 55%; ; Figure 1(b)). By adding rotenone, an inhibitor of Complex I, and succinate, a substrate for complex II, the segment comprising from complex II to complex V was evaluated. No differences in terms of oxygen uptake were observed between controls and poly(I:C) suggesting that the deficit in offspring of poly(I:C)-treated dams was located at the level of complex I. Confirming this result, complex IV activity was not different between treatments (Figure 1(b)) suggesting that mitochondrial mass was equivalent between groups. However, attempts to directly evaluate complex I activity were unsuccessful due to the limited amount of biological material.

The ratios among complexes need to be preserved to provide suitable oxidation of substrates [58]. To this end, the ratios of electron transport chain activities indicated that both treatments allow oxidizing FAD-linked substrates (such as fatty acids) similarly, whereas a significantly lower oxidation of NAD-linked substrates (such as glucose) was evident in the poly(I:C)-treated condition compared to controls (Figure 1(c)). This imbalance in the complexes’ ratios suggests that splenocytes from offspring of dams exposed to poly(I:C) use preferentially fatty acids over glucose as their main substrate for mitochondrial oxidative phosphorylation.

These results are consistent with the MD observed in lymphocytes from ASD children characterized by lower complex I activity and accompanied, in some cases, by deficits in other complexes and/or pyruvate dehydrogenase [68].

The cytokine production (IL-1β, IL-6, IL-10, IL-17, and TNF-α) from ConA-activated splenocytes obtained from adult offspring of poly(I:C)-treated animals was not different from that of SHAM-treated animals (see Supplementary Material available online on http://dx.doi.org/10.1155/2013/609602). This is consistent with the findings of others utilizing a similar MIA model in which only a handful of cytokines was still increased in early adulthood (frontal corteces IL-1α, IL-6, IL-10, and IL-9; cingulate corteces IL-10 and IFN-γ; none in hippocampus or serum [33]).

4. Conclusions

The aim of this study was to evaluate mitochondrial function in splenocytes from offspring gestationally exposed to an acute viral mimetic, that is, poly(I:C), to induce MIA. Our results indicate that the exposure of dams to a single dose of poly(I:C) at gestational day 12.5 likely triggers a TLR3-mediated response in the mother that is transmitted transplacentally to the offspring. In particular, the proinflammatory cytokine IL-6 has been proved to be a key intermediary in the behavioral changes observed in the offspring of dams treated with poly(I:C) [59]. Moreover, blocking IL-6 with antibodies prevents behavioral changes in the offspring [60], and poly(I:C)-induced MIA in IL-6 KO does not result in behavioral changes in the offspring [60]. These data seem to suggest a role for IL-6 in MIA-induced behavioral changes, although we cannot exclude other inflammatory agents such as type 1 interferons which have also been shown to take part in the response to poly(I:C) [61]. This MIA imprints a fetal programming that can still be detected during adulthood characterized by abnormal behaviors resembling those of ASD [34, 35] and SZ [34] and, biochemically, by a lower oxidative phosphorylation capacity in mitochondria within intact cells and isolated mitochondria. This suggests that prenatal immune changes ensuing the maternal poly(I:C) administration are likely to imprint the long-lasting changes in the bioenergetics of the adult offspring splenocytes. While, at the normal murine fetomaternal interface, immune cells such as neutrophils, macrophages, and NK cells are assumed to be excluded from the placenta and localized only in the decidua [62], treatment with poly(I:C) disrupts this normal distribution and induces a significant increase in the levels of proinflammatory cytokines in the placenta and a large migration of immune cells, primarily NK cells from the decidua towards the placenta, invading the spongiotrophoblast and then the labyrinth [63]. Trophoblasts, which express TLR3 [63], play a role in coordinating the maternal innate immune response to infection at the fetomaternal interface [6264] and, especially in this case, in response to viral infection.

These results beget the question, what is the link between lower complex I activity (or lower oxidative phosphorylation) in the offspring and an acute maternal immune response? A growing body of evidence is placing mitochondria at the center of bioenergetics and immune response/inflammation. Immunity to infection is also dependent on mitochondria function by regulating the synthesis of both pro- and anti-inflammatory cytokines [6569]. More recently, the view that mitochondria act as a platform facilitating innate immune responses adds to our understanding of the molecular complexity of sensor and adaptor interactions that promote effective host defense [11, 70].

Therefore, an emerging concept is that innate immune signaling is regulated by basic host metabolic functions. For instance, Toll-like receptor signaling activates mitochondrial biogenesis during critical illness [7173], perhaps in response to increased oxidative damage in host cells [71, 74, 75]. Acute inflammation is accompanied by increases in inflammatory cytokines sustained by glycolysis, whereas chronic inflammation is sustained by less inflammatory cytokines, with more reparative features fueled mainly by mitochondria-derived ATP [76]. Thus, in this study, MIA seems to imprint the immune cells of adult offspring with this more glycolytic stage resembling the influence of an acute inflammation, without switching back to the less inflammatory response. Without pointing at cause or consequence, it is interesting to note that the changes in bioenergetics in the immune cells (and not their immune response or the immune response in brain or serum [33]) segregate with the abnormal behaviors observed in this MIA model [35]. However, a number of studies have shown differential MIA induction and behavioral responses depending on the gestational exposure stage [7779]. This would suggest the existence of a window of vulnerability to infection during gestation for the onset of different behavioral defects, which may be reflected also on the mitochondrial function of the offspring.

The above effects can be explained by (i) the transfer of immune cells and/or cytokines from mother to the fetus at the maternal-fetal interface and (ii) a genetic predisposition/susceptibility of the offspring that, in association with maternal viral or bacterial infections, might increase the risk of long-lasting behavioral and immune changes [35]. Furthermore, we cannot exclude the possibility that the relatively high doses of poly(I:C) used in this study could have affected the well-being of the mother and therefore that of the fetus. For instance, poly(I:C) inhibits the development of diabetes in the NOD mouse [80] whereas the development of diabetes in diabetes-prone BB rats is poly(I:C)-dose dependent [8184]. Indeed, a recent report indicated that at least some nongenetic risk factors are shared between ASD and SZ, in particular, diabetes, exposure to drugs, nutritional deficiencies, and infectious agents among others [85].

Considering the mechanisms described above, studies are now needed to clearly identify the key players affected in this acute viral response in order to evaluate the increase in risk of either ASD or SZ that is associated with these (and other [30, 86, 87]) modifiable environmental factors to elicit public health interventions.

Authors’ Contribution

Cecilia Giulivi and Eleonora Napoli contributed equally to this work.

Acknowledgments

This study was performed under the funding from the Simons Foundation (no. 271406 to Cecilia Giulivi), NIEHS R01-ES011269, R01-ES015359, and R01-ES020392, Autism Speaks Foundation, NARSAD Foundation, Jane Botsford Johnson Foundation, and Peter Emch Foundation. The above-mentioned funding agencies were not responsible for the design and conduct of the study; collection, management, analysis, and interpretation of the data; and preparation, review, or approval of this paper. The authors of this publication declare that they have no conflicting financial interest in relation to the work described. Cecilia Giulivi, designed this study and wrote the paper; Eleonora Napoli performed all work related to mitochondrial outcomes, contributed to the writing of the paper, and analyzed the data; Jared Schwartzer and Milo Careaga performed the animal exposures, immune response, collected the cells utilized in this study, and reviewed the paper; Paul Ashwood designed the treatment of animals and contributed to the writing of the paper.

References

  1. J. Baio, “Prevalence of Autism spectrum disorders—autism and Developmental Disabilities Monitoring Network, 14 Sites, United States, 2008,” Morbidity and Mortality Weekly Report, vol. 61, no. 3, pp. 1–19, 2012. View at Google Scholar · View at Scopus
  2. I. Hertz-Picciotto and L. Delwiche, “The rise in autism and the role of age at diagnosis,” Epidemiology, vol. 20, no. 1, pp. 84–90, 2009. View at Publisher · View at Google Scholar · View at Scopus
  3. D. C. Chugani, B. S. Sundram, M. Behen, M.-L. Lee, and G. J. Moore, “Evidence of altered energy metabolism in autistic children,” Progress in Neuro-Psychopharmacology and Biological Psychiatry, vol. 23, no. 4, pp. 635–641, 1999. View at Publisher · View at Google Scholar · View at Scopus
  4. N. J. Minshew, G. Goldstein, S. M. Dombrowski, K. Panchalingam, and J. W. Pettegrew, “A preliminary 31P MRS study of autism: evidence for undersynthesis and increased degradation of brain membranes,” Biological Psychiatry, vol. 33, no. 11-12, pp. 762–773, 1993. View at Google Scholar · View at Scopus
  5. J. Lombard, “Autism: a mitochondrial disorder?” Medical Hypotheses, vol. 50, no. 6, pp. 497–500, 1998. View at Publisher · View at Google Scholar · View at Scopus
  6. C. Giulivi, Y.-F. Zhang, A. Omanska-Klusek et al., “Mitochondrial dysfunction in autism,” Journal of the American Medical Association, vol. 304, no. 21, pp. 2389–2396, 2010. View at Publisher · View at Google Scholar · View at Scopus
  7. G. Oliveira, L. Diogo, M. Grazina et al., “Mitochondrial dysfunction in autism spectrum disorders: a population-based study,” Developmental Medicine and Child Neurology, vol. 47, no. 3, pp. 185–189, 2005. View at Publisher · View at Google Scholar · View at Scopus
  8. A. Chauhan, F. Gu, M. M. Essa et al., “Brain region-specific deficit in mitochondrial electron transport chain complexes in children with autism,” Journal of Neurochemistry, vol. 117, no. 2, pp. 209–220, 2011. View at Publisher · View at Google Scholar · View at Scopus
  9. L. Ernster and G. Schatz, “Mitochondria: a historical review,” Journal of Cell Biology, vol. 91, no. 3, part 2, pp. 227s–255s, 1981. View at Google Scholar · View at Scopus
  10. A. P. West, G. S. Shadel, and S. Ghosh, “Mitochondria in innate immune responses,” Nature Reviews Immunology, vol. 11, no. 6, pp. 389–402, 2011. View at Publisher · View at Google Scholar · View at Scopus
  11. D. Arnoult, F. Soares, I. Tattoli, and S. E. Girardin, “Mitochondria in innate immunity,” EMBO Reports, vol. 12, no. 9, pp. 901–910, 2011. View at Publisher · View at Google Scholar · View at Scopus
  12. S. M. Cloonan and A. M. K. Choi, “Mitochondria: commanders of innate immunity and disease?” Current Opinion in Immunology, vol. 24, no. 1, pp. 32–40, 2012. View at Publisher · View at Google Scholar · View at Scopus
  13. G. Fossati, D. A. Moulding, D. G. Spiller, R. J. Moots, M. R. H. White, and S. W. Edwards, “The mitochondrial network of human neutrophils: role in chemotaxis, phagocytosis, respiratory burst activation, and commitment to apoptosis,” Journal of Immunology, vol. 170, no. 4, pp. 1964–1972, 2003. View at Google Scholar · View at Scopus
  14. A.-L. Genestier, M.-C. Michallet, G. Prévost et al., “Staphylococcus aureus Panton-Valentine leukocidin directly targets mitochondria and induces Bax-independent apoptosis of human neutrophils,” Journal of Clinical Investigation, vol. 115, no. 11, pp. 3117–3127, 2005. View at Publisher · View at Google Scholar · View at Scopus
  15. D. Scheel-Toellner, K. Wang, R. Craddock et al., “Reactive oxygen species limit neutrophil life span by activating death receptor signaling,” Blood, vol. 104, no. 8, pp. 2557–2564, 2004. View at Publisher · View at Google Scholar · View at Scopus
  16. N. A. Maianski, J. Geissler, S. M. Srinivasula, E. S. Alnemri, D. Roos, and T. W. Kuijpers, “Functional characterization of mitochondria in neutrophils: a role restricted to apoptosis,” Cell Death and Differentiation, vol. 11, no. 2, pp. 143–153, 2004. View at Publisher · View at Google Scholar · View at Scopus
  17. N. A. Malanski, D. Roos, and T. W. Kuijpers, “Tumor necrosis factor α induces a caspase-independent death pathway in human neutrophils,” Blood, vol. 101, no. 5, pp. 1987–1995, 2003. View at Publisher · View at Google Scholar · View at Scopus
  18. C. Burlak, A. R. Whitney, D. J. Mead, T. Hackstadt, and F. R. DeLeo, “Maturation of human neutrophil phagosomes includes incorporation of molecular chaperones and endoplasmic reticulum quality control machinery,” Molecular and Cellular Proteomics, vol. 5, no. 4, pp. 620–634, 2006. View at Publisher · View at Google Scholar · View at Scopus
  19. T. C. Pithon-Curi, M. P. De Melo, R. B. De Azevedo, T. M. T. Zorn, and R. Curi, “Glutamine utilization by rat neutrophils: presence of phosphate- dependent glutaminase,” American Journal of Physiology. Cell Physiology, vol. 273, no. 4, pp. C1124–C1129, 1997. View at Google Scholar · View at Scopus
  20. R. Curi, P. Newsholme, T. C. Pithon-Curi et al., “Metabolic fate of glutamine in lymphocytes, macrophages and neutrophils,” Brazilian Journal of Medical and Biological Research, vol. 32, no. 1, pp. 15–21, 1999. View at Google Scholar · View at Scopus
  21. K. Fukatsu, K. A. Kudsk, B. L. Zarzaur, Y. Wu, M. K. Hanna, and R. C. DeWitt, “TPN decreases IL-4 and IL-10 mRNA expression in lipopolysaccharide stimulated intestinal lamina propria cells but glutamine supplementation preserves the expression,” Shock, vol. 15, no. 4, pp. 318–322, 2001. View at Google Scholar · View at Scopus
  22. A. Yassad, A. Husson, A. Bion, and A. Lavoinne, “Synthesis of interleukin 1β and interleukin 6 by stimulated rat peritoneal macrophages: modulation by glutamine,” Cytokine, vol. 12, no. 8, pp. 1288–1291, 2000. View at Publisher · View at Google Scholar · View at Scopus
  23. P. C. Calder and P. Yaqoob, “Glutamine and the immune system,” Amino Acids, vol. 17, no. 3, pp. 227–241, 1999. View at Google Scholar · View at Scopus
  24. P. Newsholme, “Why is L-glutamine metabolism important to cells of the immune system in health, postinjury, surgery or infection?” Journal of Nutrition, vol. 131, no. 9, supplement, 2001. View at Google Scholar · View at Scopus
  25. T. C. Pithon-Curi, A. C. Levada, L. R. Lopes, S. Q. Doi, and R. Curi, “Glutamine plays a role in superoxide production and the expression of p47phox, p22phox and gp91phox in rat neutrophils,” Clinical Science, vol. 103, no. 4, pp. 403–408, 2002. View at Google Scholar · View at Scopus
  26. H. S. Hwang, S. S. Davies, M. F. Hill et al., “Role of ketoaldehyde protein adducts in ischemic cardiomyopathy,” Circulation, vol. 118, no. 18, pp. S288–S288, 2008. View at Google Scholar
  27. J. P. Gregg, L. Lit, C. A. Baron et al., “Gene expression changes in children with autism,” Genomics, vol. 91, no. 1, pp. 22–29, 2008. View at Publisher · View at Google Scholar · View at Scopus
  28. C. Onore, M. Careaga, and P. Ashwood, “The role of immune dysfunction in the pathophysiology of autism,” Brain, Behavior, and Immunity, vol. 26, no. 3, pp. 383–392, 2012. View at Publisher · View at Google Scholar · View at Scopus
  29. S. Wills, M. Cabanlit, J. Bennett, P. Ashwood, D. Amaral, and J. Van De Water, “Autoantibodies in Autism Spectrum Disorders (ASD),” Annals of the New York Academy of Sciences, vol. 1107, pp. 79–91, 2007. View at Publisher · View at Google Scholar · View at Scopus
  30. E. Napoli, C. Hung, S. Wong, and C. Giulivi, “Toxicity of the flame-retardant BDE-49 on brain mitochondria and neuronal progenitor striatal cells enhanced by a PTEN-deficient background,” Toxicological Sciences, vol. 132, no. 1, pp. 196–210, 2013. View at Google Scholar
  31. A. L. Burgueno, R. Cabrerizo, M. N. Gonzales, S. Sookoian, and C. J. Pirola, “Maternal high-fat intake during pregnancy programs metabolic-syndrome-related phenotypes through liver mitochondrial DNA copy number and transcriptional activity of liver PPARGC1A,” Journal of Nutritional Biochemistry, vol. 24, no. 1, pp. 6–13, 2013. View at Google Scholar
  32. A. P. Garcia, T. Priego, M. Palou, J. Sanchez, A. Palou, and C. Pico, “Early alterations in plasma ghrelin levels in offspring of calorie-restricted rats during gestation may be linked to lower sympathetic drive to the stomach,” Peptides, vol. 39, pp. 59–63, 2013. View at Google Scholar
  33. P. A. Garay, E. Y. Hsiao, P. H. Patterson, and A. K. McAllister, “Maternal immune activation causes age- and region-specific changes in brain cytokines in offspring throughout development,” Brain, Behavior, and Immunity, vol. 31, pp. 54–68, 2013. View at Google Scholar
  34. P. H. Patterson, “Modeling autistic features in animals,” Pediatric Research, vol. 69, no. 5, part 2, pp. 34R–40R, 2011. View at Publisher · View at Google Scholar · View at Scopus
  35. J. J. Schwartzer, M. Careaga, C. E. Onore, J. A. Rushakoff, R. F. Berman, and P. Ashwood, “Maternal immune activation and strain specific interactions in the development of autism-like behaviors in mice,” Translational Psychiatry, vol. 3, article e240, 2013. View at Google Scholar
  36. J. H. Tinsley, V. L. Chiasson, A. Mahajan, K. J. Young, and B. M. Mitchell, “Toll-like receptor 3 activation during pregnancy elicits preeclampsia-like symptoms in rats,” American Journal of Hypertension, vol. 22, no. 12, pp. 1314–1319, 2009. View at Publisher · View at Google Scholar · View at Scopus
  37. P. Chatterjee, V. L. Chiasson, S. E. Kopriva et al., “Interleukin 10 deficiency exacerbates toll-like receptor 3-induced preeclampsia-like symptoms in mice,” Hypertension, vol. 58, no. 3, pp. 489–496, 2011. View at Publisher · View at Google Scholar · View at Scopus
  38. M. J. Cipolla, E. M. Houston, R. P. Kraig, and E. A. Bonney, “Differential effects of low-dose endotoxin on the cerebral circulation during pregnancy,” Reproductive Sciences, vol. 18, no. 12, pp. 1211–1221, 2011. View at Publisher · View at Google Scholar · View at Scopus
  39. V. M. Abrahams, P. Bole-Aldo, Y. M. Kim et al., “Divergent trophoblast responses to bacterial products mediated by TLRs,” Journal of Immunology, vol. 173, no. 7, pp. 4286–4296, 2004. View at Google Scholar · View at Scopus
  40. A. Pineda, S. L. Verdin-Terán, A. Camacho, and L. Moreno-Fierros, “Expression of toll-like receptor TLR-2, TLR-3, TLR-4 and TLR-9 is increased in placentas from patients with preeclampsia,” Archives of Medical Research, vol. 42, no. 5, pp. 382–391, 2011. View at Publisher · View at Google Scholar · View at Scopus
  41. C. K. Walker, K. W. Anderson, K. M. Milano et al., “Trophoblast inclusions are significantly increased in the placentas of children in families at risk for autism,” Biological Psychiatry, vol. 74, no. 3, pp. 204–211, 2013. View at Publisher · View at Google Scholar
  42. A. S. Brown and E. J. Derkits, “Prenatal infection and schizophrenia: a review of epidemiologic and translational studies,” American Journal of Psychiatry, vol. 167, no. 3, pp. 261–280, 2010. View at Publisher · View at Google Scholar · View at Scopus
  43. A. S. Brown and P. H. Patterson, “Maternal infection and schizophrenia: implications for prevention,” Schizophrenia Bulletin, vol. 37, no. 2, pp. 284–290, 2011. View at Publisher · View at Google Scholar · View at Scopus
  44. P. H. Patterson, “Maternal infection and immune involvement in autism,” Trends in Molecular Medicine, vol. 17, no. 7, pp. 389–394, 2011. View at Publisher · View at Google Scholar · View at Scopus
  45. P. H. Patterson, “Maternal infection and autism,” Brain, Behavior, and Immunity, vol. 26, no. 3, p. 393, 2012. View at Publisher · View at Google Scholar · View at Scopus
  46. A. S. Brown, M. D. Begg, S. Gravenstein et al., “Serologic evidence of prenatal influenza in the etiology of schizophrenia,” Archives of General Psychiatry, vol. 61, no. 8, pp. 774–780, 2004. View at Publisher · View at Google Scholar · View at Scopus
  47. A. S. Brown, J. Hooton, C. A. Schaefer et al., “Elevated maternal interleukin-8 levels and risk of schizophrenia in adult offspring,” American Journal of Psychiatry, vol. 161, no. 5, pp. 889–895, 2004. View at Publisher · View at Google Scholar · View at Scopus
  48. H. Ó. Atladóttir, P. Thorsen, L. Østergaard et al., “Maternal infection requiring hospitalization during pregnancy and autism spectrum disorders,” Journal of Autism and Developmental Disorders, vol. 40, no. 12, pp. 1423–1430, 2010. View at Publisher · View at Google Scholar · View at Scopus
  49. J. H. Gilmore, L. F. Jarskog, S. Vadlamudi, and J. M. Lauder, “Prenatal infection and risk for schizophrenia: IL-1β, IL-6, and TNFα inhibit cortical neuron dendrite development,” Neuropsychopharmacology, vol. 29, no. 7, pp. 1221–1229, 2004. View at Publisher · View at Google Scholar · View at Scopus
  50. U. Meyer, J. Feldon, M. Schedlowski, and B. K. Yee, “Towards an immuno-precipitated neurodevelopmental animal model of schizophrenia,” Neuroscience and Biobehavioral Reviews, vol. 29, no. 6, pp. 913–947, 2005. View at Publisher · View at Google Scholar · View at Scopus
  51. B. E. Deverman and P. H. Patterson, “Cytokines and CNS Development,” Neuron, vol. 64, no. 1, pp. 61–78, 2009. View at Publisher · View at Google Scholar · View at Scopus
  52. P. A. Garay and A. K. McAllister, “Novel roles for immune molecules in neural development: implications for neurodevelopmental disorders,” Frontiers in Synaptic Neuroscience, vol. 2, article 136, 2010. View at Google Scholar
  53. G. Arrode-Bruses and J. L. Bruses, “Maternal immune activation by poly I:C induces expression of cytokines IL-1beta and IL-13, chemokine MCP-1 and colony stimulating factor VEGF in fetal mouse brain,” Journal of Neuroinflammation, vol. 9, article 83, 2012. View at Google Scholar
  54. A. Burton, O. Kizhner, M. B. Brown, and M. R. Peltier, “Effect of experimental genital mycoplasmosis on gene expression in the fetal brain,” Journal of Reproductive Immunology, vol. 93, no. 1, pp. 9–16, 2012. View at Publisher · View at Google Scholar · View at Scopus
  55. B. Clancy, B. L. Finlay, R. B. Darlington, and K. J. S. Anand, “Extrapolating brain development from experimental species to humans,” NeuroToxicology, vol. 28, no. 5, pp. 931–937, 2007. View at Publisher · View at Google Scholar · View at Scopus
  56. C. M. Jewell, S. C. Lopez, and D. J. Irvine, “In situ engineering of the lymph node microenvironment via intranodal injection of adjuvant-releasing polymer particles,” Proceedings of the National Academy of Sciences of the United States of America, vol. 108, no. 38, pp. 15745–15750, 2011. View at Publisher · View at Google Scholar · View at Scopus
  57. P. Rustin, D. Chretien, T. Bourgeron et al., “Biochemical and molecular investigations in respiratory chain deficiencies,” Clinica Chimica Acta, vol. 228, no. 1, pp. 35–51, 1994. View at Publisher · View at Google Scholar · View at Scopus
  58. Y. Hatefi, “The mitochondrial electron transport and oxidative phosphorylation system,” Annual Review of Biochemistry, vol. 54, pp. 1015–1069, 1985. View at Google Scholar · View at Scopus
  59. M. Mandal, A. C. Marzouk, R. Donnelly, and N. M. Ponzio, “Maternal immune stimulation during pregnancy affects adaptive immunity in offspring to promote development of TH17 cells,” Brain, Behavior, and Immunity, vol. 25, no. 5, pp. 863–871, 2011. View at Publisher · View at Google Scholar · View at Scopus
  60. S. E. P. Smith, J. Li, K. Garbett, K. Mirnics, and P. H. Patterson, “Maternal immune activation alters fetal brain development through interleukin-6,” Journal of Neuroscience, vol. 27, no. 40, pp. 10695–10702, 2007. View at Publisher · View at Google Scholar · View at Scopus
  61. G. A. Farina, M. R. York, M. Di Marzio et al., “Poly(I:C) drives type i IFN- and TGFΒ-mediated inflammation and dermal fibrosis simulating altered gene expression in systemic sclerosis,” Journal of Investigative Dermatology, vol. 130, no. 11, pp. 2583–2593, 2010. View at Publisher · View at Google Scholar · View at Scopus
  62. I. Guleria and J. W. Pollard, “The trophoblast is a component of the innate immune system during pregnancy,” Nature Medicine, vol. 6, no. 5, pp. 589–593, 2000. View at Publisher · View at Google Scholar · View at Scopus
  63. K. Koga, I. Cardenas, P. Aldo et al., “Activation of TLR3 in the trophoblast is associated with preterm delivery,” American Journal of Reproductive Immunology, vol. 61, no. 3, pp. 196–212, 2009. View at Publisher · View at Google Scholar · View at Scopus
  64. G. Mor, R. Romero, P. B. Aldo, and V. M. Abrahams, “Is the trophoblast an immune regulator? The role of toll-like receptors during pregnancy,” Critical Reviews in Immunology, vol. 25, no. 5, pp. 375–388, 2005. View at Publisher · View at Google Scholar · View at Scopus
  65. H. B. Suliman, K. E. Welty-Wolf, M. S. Carraway, D. A. Schwartz, J. W. Hollingsworth, and C. A. Piantadosi, “Toll-like receptor 4 mediates mitochondrial DNA damage and biogenic responses after heat-inactivated E. coli,” The FASEB Journal, vol. 19, no. 11, pp. 1531–1533, 2005. View at Publisher · View at Google Scholar · View at Scopus
  66. M. Sasai, M. Shingai, K. Funami et al., “NAK-associated protein 1 participates in both the TLR3 and the cytoplasmic pathways in type I IFN induction,” Journal of Immunology, vol. 177, no. 12, pp. 8676–8683, 2006. View at Google Scholar · View at Scopus
  67. R. B. Seth, L. Sun, and Z. J. Chen, “Antiviral innate immunity pathways,” Cell Research, vol. 16, no. 2, pp. 141–147, 2006. View at Publisher · View at Google Scholar · View at Scopus
  68. H.-X. Shi, X. Liu, Q. Wang et al., “Mitochondrial ubiquitin ligase MARCH5 promotes TLR7 signaling by attenuating TANK action,” PLoS Pathogens, vol. 7, no. 5, Article ID e1002057, 2011. View at Publisher · View at Google Scholar · View at Scopus
  69. S. Carta, S. Tassi, L. Delfino et al., “Deficient production of IL-1 receptor antagonist and IL-6 coupled to oxidative stress in cryopyrin-associated periodic syndrome monocytes,” Annals of the Rheumatic Diseases, vol. 71, no. 9, pp. 1577–1581, 2012. View at Google Scholar
  70. N. Subramanian, K. Natarajan, M. R. Clatworthy, Z. Wang, and R. N. Germain, “The adaptor MAVS promotes NLRP3 mitochondrial localization and inflammasome activation,” Cell, vol. 153, no. 2, pp. 348–361, 2013. View at Google Scholar
  71. T. E. Sweeney, H. B. Suliman, J. W. Hollingsworth, K. E. Welty-Wolf, and C. A. Piantadosi, “A toll-like receptor 2 pathway regulates the Ppargc1a/b metabolic co-activators in mice with Staphylococcal aureus sepsis,” PLoS One, vol. 6, no. 9, Article ID e25249, 2011. View at Publisher · View at Google Scholar · View at Scopus
  72. C. A. Piantadosi and H. B. Suliman, “Transcriptional control of mitochondrial biogenesis and its interface with inflammatory processes,” Biochimica et Biophysica Acta, vol. 1820, no. 4, pp. 532–541, 2012. View at Publisher · View at Google Scholar · View at Scopus
  73. C. P. Bauerfeld, R. Rastogi, G. Pirockinaite et al., “TLR4-mediated AKT activation is Myd88/TRIF dependent and critical for induction of oxidative phosphorylation and mitochondrial transcription factor a in murine macrophages,” Journal of Immunology, vol. 188, no. 6, pp. 2847–2857, 2012. View at Publisher · View at Google Scholar · View at Scopus
  74. T. E. Sweeney, H. B. Suliman, J. W. Hollingsworth, and C. A. Piantadosi, “Differential regulation of the PGC family of genes in a mouse model of staphylococcus aureus sepsis,” PLoS One, vol. 5, no. 7, Article ID e11606, 2010. View at Publisher · View at Google Scholar · View at Scopus
  75. T. Oka, S. Hikoso, O. Yamaguchi et al., “Mitochondrial DNA that escapes from autophagy causes inflammation and heart failure,” Nature, vol. 485, no. 7397, pp. 251–255, 2012. View at Google Scholar
  76. T. F. Liu, C. M. Brown, M. El Gazzar et al., “Fueling the flame: bioenergy couples metabolism and inflammation,” Journal of Leukocyte Biology, vol. 92, no. 3, pp. 499–507, 2012. View at Google Scholar
  77. M.-E. Fortier, G. N. Luheshi, and P. Boksa, “Effects of prenatal infection on prepulse inhibition in the rat depend on the nature of the infectious agent and the stage of pregnancy,” Behavioural Brain Research, vol. 181, no. 2, pp. 270–277, 2007. View at Publisher · View at Google Scholar · View at Scopus
  78. U. Meyer, M. Nyffeler, A. Engler et al., “The time of prenatal immune challenge determines the specificity of inflammation-mediated brain and behavioral pathology,” Journal of Neuroscience, vol. 26, no. 18, pp. 4752–4762, 2006. View at Publisher · View at Google Scholar · View at Scopus
  79. P. H. Patterson, “Immune involvement in schizophrenia and autism: etiology, pathology and animal models,” Behavioural Brain Research, vol. 204, no. 2, pp. 313–321, 2009. View at Publisher · View at Google Scholar · View at Scopus
  80. D. V. Serreze, K. Hamaguchi, and E. H. Leiter, “Immunostimulation circumvents diabetes in NOD Lt mice,” Journal of Autoimmunity, vol. 2, no. 6, pp. 759–776, 1989. View at Google Scholar · View at Scopus
  81. D. O. Sobel, D. Goyal, B. Ahvazi et al., “Low dose poly I:C prevents diabetes in the diabetes prone BB rat,” Journal of Autoimmunity, vol. 11, no. 4, pp. 343–352, 1998. View at Publisher · View at Google Scholar · View at Scopus
  82. V. A. Thomas, B. A. Woda, E. S. Handler, D. L. Greiner, J. P. Mordes, and A. A. Rossini, “Altered expression of diabetes in BB/Wor rats by exposure to viral pathogens,” Diabetes, vol. 40, no. 2, pp. 255–258, 1991. View at Google Scholar · View at Scopus
  83. C. H. Ewel, D. O. Sobel, B. J. Zeligs, and J. A. Bellanti, “Poly I:C accelerates development of diabetes mellitus in diabetes-prone BB rat,” Diabetes, vol. 41, no. 8, pp. 1016–1021, 1992. View at Google Scholar · View at Scopus
  84. D. O. Sobel, J. Newsome, C. H. Ewel et al., “Poly I:C induces development of diabetes mellitus in BB rat,” Diabetes, vol. 41, no. 4, pp. 515–520, 1992. View at Google Scholar · View at Scopus
  85. J. Hamlyn, M. Duhig, J. McGrath, and J. Scott, “Modifiable risk factors for schizophrenia and autism–shared risk factors impacting on brain development,” Neurobiology of Disease, vol. 53, pp. 3–9, 2013. View at Google Scholar
  86. E. Napoli, C. Ross-Inta, S. Wong et al., “Mitochondrial dysfunction in Pten haplo-insufficient mice with social deficits and repetitive behavior: interplay between Pten and p53,” PLoS One, vol. 7, no. 8, Article ID e42504, 2012. View at Google Scholar
  87. E. Napoli, S. Wong, and C. Giulivi, “Evidence of reactive oxygen species-mediated damage to mitochondrial DNA in children with typical autism,” Molecular Autism, vol. 4, no. 1, article 2, 2013. View at Publisher · View at Google Scholar