www.sciencemag.org/cgi/content/full/science.aad0314/DC1 Supplementary Materials for The maternal interleukin-17a pathway in mice promotes autism-like phenotypes in offspring Gloria B. Choi, Yeong S. Yim, Helen Wong, Sangdoo Kim, Hyunju Kim, Sangwon V. Kim, Charles A. Hoeffer*, Dan R. Littman*, Jun R. Huh* *Corresponding author. E-mail: [email protected] (C.A.H.); [email protected](D.R.L.); [email protected] (J.R.H.) Published 28 January 2016 on Science Express DOI: 10.1126/science.aad0314 This PDF file includes Materials and Methods Supplementary Text Figs. S1 to S11 References
35
Embed
Supplementary Materials for - Sciencescience.sciencemag.org/content/sci/suppl/2016/01/27/science.aad... · Supplementary Materials for The maternal interleukin-17a pathway in mice
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
C.A.H., H.W., G.B.C., Y.S.Y., J.R.H. and D.R.L. designed the experiments and/or provided advice and technical expertise. G.B.C., Y.S.Y., H.W., S.K., H.K., C.A.H. and J.R.H., performed the experiments. S.V.K. generated RORt conditional mouse lines. J.R.H., G.B.C., Y.S.Y., H.W., C.A.H. and D.R.L. wrote the manuscript with input from the co-authors.
16
Fig. S1. Expression of multiple cytokines detected upon MIA. (A, B, C) Maternal
serum concentrations of TNF-, IFN- and IL-1 (n=3-6 mice per group, pooled from
two independent experiments) at 3, 24, 48 or 96 h after PBS or poly(I:C) injection of
pregnant dams. (D) Serum and placenta/decidua concentrations of IL-10 at E15.5
(n=5~10 mice per group). (E) Serum concentrations of maternal IL-17a (n=5-8 mice per
group, pooled from two independent experiments) at E14.5 in WT or IL-6 KO mothers
injected with PBS, recombinant IL-6 (mIL-6), or poly(I:C). (F) Supernatant
concentrations of IL-17a from ex vivo cultured mononuclear cells, isolated from
duodenum of PBS- or poly(I:C)-treated pregnant dams. Stim refers to PMA and
HET), n=28 (Poly(I:C), IL-17Ra-KO)). (B) Serum concentrations of maternal IL-17a
(n=5~8 mice per group, pooled from two independent experiments) at E14.5 in PBS- or
poly(I:C)-injected WT or IL-17Ra HET mothers. (A) Two-way ANOVA with Tukey
post-hoc testing. (B) One-way ANOVA with Tukey post-hoc testing. ***P < 0.001 and
**P < 0.01. Graphs show mean +/- s.e.m.
26
Fig. S9. Characterization of the disorganized cortical patch from intra-ventricular
administration of IL-17a. (A) SATB2 and TBR1 staining of E18.5 fetal brains from
animals treated as in (Fig. 4A). Images are representative of five independent
experiments. (B) Thickness of the cortical plate in E18.5 fetal brains. (A and B) (i), (ii)
and (iii) indicate subdivisions resulting from equally dividing the cortex perpendicularly
through the cortical plate. Scale bar represents 100 m. (B) Student’s t test. **P < 0.01,
*P < 0.05, and ns; not significant. Graphs show mean +/- s.e.m.
27
Fig. S10. IL-17a acts downstream of IL-6 in the MIA model. (A) SATB2 and TBR1
staining of the cortex in E18.5 fetal brains. PBS, IL-6 or IL-17a were intraventricularly
injected into the fetal brain of the indicated genotypes at E14.5. Images are representative
of 2-3 independent experiments. (B) Quantification of TBR1 and SATB2 positive cells in
a 300x300 m2 ROI centered on the malformation in the cortical plate (n=6 (PBS, WT
dam), n=6 (IL-6, WT dam), n=6 (IL-17a, IL-17Ra KO dam); from 2-3 independent dams
per treatment). (C) Ultrasonic vocalization (USV) assay. At P9, pups from the indicated
conditions in (A) were separated from their mothers to elicit USV calls. The number of
pup calls is plotted on the y-axis (n=10 (PBS, WT dam), n=14 (IL-6, WT dam), n=20
(IL-17a, IL-17Ra KO dam); from 2-3 independent dams per treatment). (D) SATB2 and
28
TBR1 staining in the cortex of E18.5 fetal brain, derived from PBS- or IL-6-injected
mothers, pretreated with isotype control (Cont) or IL-17a blocking antibodies (anti-IL-
17a). (E) Quantification of TBR1 and SATB2 positive cells in a 300x300 m2 ROI
centered on the cortical plate containing the cortical patch (n=6 (PBS, Cont), n=6 (IL-6,
Cont), n=6 (IL-6, anti-IL-17a); from 2-3 independent dams per treatment). (F) USV assay
for the pups from the indicated conditions as (D) (n=10 (PBS, Cont), n=19 (IL-6, Cont),
n=18 (IL-6, anti-IL-17a); from 3-4 independent dams per treatment). (G) USV assay for
the pups injected with PBS or IL-17a and derived from IL-6 KO mothers injected with
Poly(I:C) (n=8 (PBS,Poly(I:C)), n=10 (IL-17a,Poly(I:C)); from 2 independent dams per
treatment). (B and E) Two-way ANOVA with Tukey post-hoc tests. (C and F) One-way
ANOVA with Tukey post-hoc tests. (G) Student’s t test. **P < 0.01, *P < 0.05 and ns;
not significant. Graphs show mean +/- s.e.m.
29
Fig. S11. A proposed mechanism by which maternal Th17 cells and IL-17a induce
MIA-dependent behavioral and cortical abnormalities in offspring.
30
REFERENCES AND NOTES
1. H. O. Atladóttir, P. Thorsen, L. Østergaard, D. E. Schendel, S. Lemcke, M. Abdallah, E. T. Parner, Maternal infection requiring hospitalization during pregnancy and autism spectrum disorders. J. Autism Dev. Disord. 40, 1423–1430 (2010). Medline doi:10.1007/s10803-010-1006-y
2. P. H. Patterson, Immune involvement in schizophrenia and autism: Etiology, pathology and animal models. Behav. Brain Res. 204, 313–321 (2009). Medline doi:10.1016/j.bbr.2008.12.016
3. A. S. Brown, A. Sourander, S. Hinkka-Yli-Salomäki, I. W. McKeague, J. Sundvall, H. M. Surcel, Elevated maternal C-reactive protein and autism in a national birth cohort. Mol. Psychiatry 19, 259–264 (2014). Medline doi:10.1038/mp.2012.197
4. H. O. Atladóttir, M. G. Pedersen, P. Thorsen, P. B. Mortensen, B. Deleuran, W. W. Eaton, E. T. Parner, Association of family history of autoimmune diseases and autism spectrum disorders. Pediatrics 124, 687–694 (2009). Medline doi:10.1542/peds.2008-2445
5. P. Ashwood, S. Wills, J. Van de Water, The immune response in autism: A new frontier for autism research. J. Leukoc. Biol. 80, 1–15 (2006). Medline doi:10.1189/jlb.1205707
6. B. K. Lee, C. Magnusson, R. M. Gardner, Å. Blomström, C. J. Newschaffer, I. Burstyn, H. Karlsson, C. Dalman, Maternal hospitalization with infection during pregnancy and risk of autism spectrum disorders. Brain Behav. Immun. 44, 100–105 (2015). Medline doi:10.1016/j.bbi.2014.09.001
7. S. E. P. Smith, J. Li, K. Garbett, K. Mirnics, P. H. Patterson, Maternal immune activation alters fetal brain development through interleukin-6. J. Neurosci. 27, 10695–10702 (2007).doi:10.1523/JNEUROSCI.2178-07.2007 Medline
8. N. V. Malkova, C. Z. Yu, E. Y. Hsiao, M. J. Moore, P. H. Patterson, Maternal immune activation yields offspring displaying mouse versions of the three core symptoms of autism. Brain Behav. Immun. 26, 607–616 (2012). Medline doi:10.1016/j.bbi.2012.01.011
9. C. M. Wilke, K. Bishop, D. Fox, W. Zou, Deciphering the role of Th17 cells in human disease. Trends Immunol. 32, 603–611 (2011). Medline doi:10.1016/j.it.2011.08.003
10. N. Manel, D. Unutmaz, D. R. Littman, The differentiation of human TH-17 cells requires transforming growth factor-beta and induction of the nuclear receptor RORgammat. Nat. Immunol. 9, 641–649 (2008). Medline doi:10.1038/ni.1610
11. H. Spits, J. P. Di Santo, The expanding family of innate lymphoid cells: Regulators and effectors of immunity and tissue remodeling. Nat. Immunol. 12, 21–27 (2011). Medline doi:10.1038/ni.1962
12. M. Lochner, L. Peduto, M. Cherrier, S. Sawa, F. Langa, R. Varona, D. Riethmacher, M. Si-Tahar, J. P. Di Santo, G. Eberl, In vivo equilibrium of proinflammatory IL-
31
17+ and regulatory IL-10+ Foxp3+ RORgamma t+ T cells. J. Exp. Med. 205, 1381–1393 (2008). Medline doi:10.1084/jem.20080034
13. I. I. Ivanov, B. S. McKenzie, L. Zhou, C. E. Tadokoro, A. Lepelley, J. J. Lafaille, D. J. Cua, D. R. Littman, The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell 126, 1121–1133 (2006). Medline doi:10.1016/j.cell.2006.07.035
14. L. Y. Al-Ayadhi, G. A. Mostafa, Elevated serum levels of interleukin-17A in children with autism. J. Neuroinflammation 9, 158 (2012). Medline doi:10.1186/1742-2094-9-158
15. K. Suzuki, H. Matsuzaki, K. Iwata, Y. Kameno, C. Shimmura, S. Kawai, Y. Yoshihara, T. Wakuda, K. Takebayashi, S. Takagai, K. Matsumoto, K. J. Tsuchiya, Y. Iwata, K. Nakamura, M. Tsujii, T. Sugiyama, N. Mori, Plasma cytokine profiles in subjects with high-functioning autism spectrum disorders. PLOS ONE 6, e20470 (2011). Medline doi:10.1371/journal.pone.0020470
16. B. van der Zwaag, L. Franke, M. Poot, R. Hochstenbach, H. A. Spierenburg, J. A. Vorstman, E. van Daalen, M. V. de Jonge, N. E. Verbeek, E. H. Brilstra, R. van ’t Slot, R. A. Ophoff, M. A. van Es, H. M. Blauw, J. H. Veldink, J. E. Buizer-Voskamp, F. A. Beemer, L. H. van den Berg, C. Wijmenga, H. K. van Amstel, H. van Engeland, J. P. Burbach, W. G. Staal, Gene-network analysis identifies susceptibility genes related to glycobiology in autism. PLOS ONE 4, e5324 (2009). Medline doi:10.1371/journal.pone.0005324
17. M. Mandal, A. C. Marzouk, R. Donnelly, N. M. Ponzio, Preferential development of Th17 cells in offspring of immunostimulated pregnant mice. J. Reprod. Immunol. 87, 97–100 (2010). Medline doi:10.1016/j.jri.2010.06.156
18. E. Y. Hsiao, S. W. McBride, J. Chow, S. K. Mazmanian, P. H. Patterson, Modeling an autism risk factor in mice leads to permanent immune dysregulation. Proc. Natl. Acad. Sci. U.S.A. 109, 12776–12781 (2012). Medline doi:10.1073/pnas.1202556109
19. V. K. Kuchroo, A. Awasthi, Emerging new roles of Th17 cells. Eur. J. Immunol. 42, 2211–2214 (2012). Medline doi:10.1002/eji.201242872
20. C. Dehay, H. Kennedy, Cell-cycle control and cortical development. Nat. Rev. Neurosci. 8, 438–450 (2007). Medline doi:10.1038/nrn2097
21. M. F. Casanova, A. S. El-Baz, S. S. Kamat, B. A. Dombroski, F. Khalifa, A. Elnakib, A. Soliman, A. Allison-McNutt, A. E. Switala, Focal cortical dysplasias in autism spectrum disorders. Acta Neuropathol. Commun. 1, 67 (2013). Medline doi:10.1186/2051-5960-1-67
22. R. Stoner, M. L. Chow, M. P. Boyle, S. M. Sunkin, P. R. Mouton, S. Roy, A. Wynshaw-Boris, S. A. Colamarino, E. S. Lein, E. Courchesne, Patches of disorganization in the neocortex of children with autism. N. Engl. J. Med. 370, 1209–1219 (2014). Medline doi:10.1056/NEJMoa1307491
23. J. De Miranda, K. Yaddanapudi, M. Hornig, G. Villar, R. Serge, W. I. Lipkin, Induction of Toll-like receptor 3-mediated immunity during gestation inhibits
24. S. E. Smith, R. M. Elliott, M. P. Anderson, Maternal immune activation increases neonatal mouse cortex thickness and cell density. J. Neuroimmune Pharmacol. 7, 529–532 (2012). Medline doi:10.1007/s11481-012-9372-1
25. B. J. Molyneaux, P. Arlotta, J. R. Menezes, J. D. Macklis, Neuronal subtype specification in the cerebral cortex. Nat. Rev. Neurosci. 8, 427–437 (2007). Medline doi:10.1038/nrn2151
26. E. A. Alcamo, L. Chirivella, M. Dautzenberg, G. Dobreva, I. Fariñas, R. Grosschedl, S. K. McConnell, Satb2 regulates callosal projection neuron identity in the developing cerebral cortex. Neuron 57, 364–377 (2008). Medline doi:10.1016/j.neuron.2007.12.012
27. C. Englund, A. Fink, C. Lau, D. Pham, R. A. Daza, A. Bulfone, T. Kowalczyk, R. F. Hevner, Pax6, Tbr2, and Tbr1 are expressed sequentially by radial glia, intermediate progenitor cells, and postmitotic neurons in developing neocortex. J. Neurosci. 25, 247–251 (2005). Medline doi:10.1523/JNEUROSCI.2899-04.2005
28. M. Leid, J. E. Ishmael, D. Avram, D. Shepherd, V. Fraulob, P. Dollé, CTIP1 and CTIP2 are differentially expressed during mouse embryogenesis. Gene Expr. Patterns 4, 733–739 (2004). Medline doi:10.1016/j.modgep.2004.03.009
29. J. J. Schwartzer, M. Careaga, C. E. Onore, J. A. Rushakoff, R. F. Berman, P. Ashwood, Maternal immune activation and strain specific interactions in the development of autism-like behaviors in mice. Transl. Psychiatry 3, e240 (2013). Medline doi:10.1038/tp.2013.16
30. N. Yee, R. K. Schwarting, E. Fuchs, M. Wöhr, Increased affective ultrasonic communication during fear learning in adult male rats exposed to maternal immune activation. J. Psychiatr. Res. 46, 1199–1205 (2012). Medline doi:10.1016/j.jpsychires.2012.05.010
31. E. Y. Hsiao, S. W. McBride, S. Hsien, G. Sharon, E. R. Hyde, T. McCue, J. A. Codelli, J. Chow, S. E. Reisman, J. F. Petrosino, P. H. Patterson, S. K. Mazmanian, Microbiota modulate behavioral and physiological abnormalities associated with neurodevelopmental disorders. Cell 155, 1451–1463 (2013). Medline doi:10.1016/j.cell.2013.11.024
32. C. A. Hoeffer, W. Tang, H. Wong, A. Santillan, R. J. Patterson, L. A. Martinez, M. V. Tejada-Simon, R. Paylor, S. L. Hamilton, E. Klann, Removal of FKBP12 enhances mTOR-Raptor interactions, LTP, memory, and perseverative/repetitive behavior. Neuron 60, 832–845 (2008). Medline doi:10.1016/j.neuron.2008.09.037
33. H. X. Wu, L. P. Jin, B. Xu, S. S. Liang, D. J. Li, Decidual stromal cells recruit Th17 cells into decidua to promote proliferation and invasion of human trophoblast cells by secreting IL-17. Cell. Mol. Immunol. 11, 253–262 (2014). Medline doi:10.1038/cmi.2013.67
33
34. A. Nakashima, M. Ito, S. Yoneda, A. Shiozaki, T. Hidaka, S. Saito, Circulating and decidual Th17 cell levels in healthy pregnancy. Am. J. Reprod. Immunol. 63, 104–109 (2010). Medline doi:10.1111/j.1600-0897.2009.00771.x
35. E. A. Martínez-García, B. Chávez-Robles, P. E. Sánchez-Hernández, L. Núñez-Atahualpa, B. T. Martín-Máquez, A. Muñoz-Gómez, L. González-López, J. I. Gámez-Nava, M. Salazar-Páramo, I. Dávalos-Rodríguez, M. H. Petri, D. Zúñiga-Tamayo, R. Vargas-Ramírez, M. Vázquez-Del Mercado, IL-17 increased in the third trimester in healthy women with term labor. Am. J. Reprod. Immunol. 65, 99–103 (2011). Medline doi:10.1111/j.1600-0897.2010.00893.x
36. G. Eberl, D. R. Littman, Thymic origin of intestinal alphabeta T cells revealed by fate mapping of RORgammat+ cells. Science 305, 248–251 (2004). Medline doi:10.1126/science.1096472
37. Z. Sun, D. Unutmaz, Y. R. Zou, M. J. Sunshine, A. Pierani, S. Brenner-Morton, R. E. Mebius, D. R. Littman, Requirement for RORgamma in thymocyte survival and lymphoid organ development. Science 288, 2369–2373 (2000). Medline doi:10.1126/science.288.5475.2369
38. J. R. Huh, M. W. Leung, P. Huang, D. A. Ryan, M. R. Krout, R. R. Malapaka, J. Chow, N. Manel, M. Ciofani, S. V. Kim, A. Cuesta, F. R. Santori, J. J. Lafaille, H. E. Xu, D. Y. Gin, F. Rastinejad, D. R. Littman, Digoxin and its derivatives suppress TH17 cell differentiation by antagonizing RORγt activity. Nature 472, 486–490 (2011). Medline doi:10.1038/nature09978
39. Y. Iwakura, H. Ishigame, S. Saijo, S. Nakae, Functional specialization of interleukin-17 family members. Immunity 34, 149–162 (2011). Medline doi:10.1016/j.immuni.2011.02.012
40. E. Y. Hsiao, P. H. Patterson, Activation of the maternal immune system induces endocrine changes in the placenta via IL-6. Brain Behav. Immun. 25, 604–615 (2011). Medline doi:10.1016/j.bbi.2010.12.017
41. L. A. Orosco, A. P. Ross, S. L. Cates, S. E. Scott, D. Wu, J. Sohn, D. Pleasure, S. J. Pleasure, I. E. Adamopoulos, K. S. Zarbalis, Loss of Wdfy3 in mice alters cerebral cortical neurogenesis reflecting aspects of the autism pathology. Nat. Commun. 5, 4692 (2014). Medline doi:10.1038/ncomms5692
42. O. Peñagarikano, B. S. Abrahams, E. I. Herman, K. D. Winden, A. Gdalyahu, H. Dong, L. I. Sonnenblick, R. Gruver, J. Almajano, A. Bragin, P. Golshani, J. T. Trachtenberg, E. Peles, D. H. Geschwind, Absence of CNTNAP2 leads to epilepsy, neuronal migration abnormalities, and core autism-related deficits. Cell 147, 235–246 (2011). Medline doi:10.1016/j.cell.2011.08.040
43. S. G. Hymowitz, E. H. Filvaroff, J. P. Yin, J. Lee, L. Cai, P. Risser, M. Maruoka, W. Mao, J. Foster, R. F. Kelley, G. Pan, A. L. Gurney, A. M. de Vos, M. A. Starovasnik, IL-17s adopt a cystine knot fold: Structure and activity of a novel cytokine, IL-17F, and implications for receptor binding. EMBO J. 20, 5332–5341 (2001). Medline doi:10.1093/emboj/20.19.5332
34
44. X. Zhang, P. Angkasekwinai, C. Dong, H. Tang, Structure and function of interleukin-17 family cytokines. Protein Cell 2, 26–40 (2011). Medline doi:10.1007/s13238-011-1006-5
45. K. Chen, J. P. McAleer, Y. Lin, D. L. Paterson, M. Zheng, J. F. Alcorn, C. T. Weaver, J. K. Kolls, Th17 cells mediate clade-specific, serotype-independent mucosal immunity. Immunity 35, 997–1009 (2011). Medline doi:10.1016/j.immuni.2011.10.018
46. I. I. Ivanov, K. Atarashi, N. Manel, E. L. Brodie, T. Shima, U. Karaoz, D. Wei, K. C. Goldfarb, C. A. Santee, S. V. Lynch, T. Tanoue, A. Imaoka, K. Itoh, K. Takeda, Y. Umesaki, K. Honda, D. R. Littman, Induction of intestinal Th17 cells by segmented filamentous bacteria. Cell 139, 485–498 (2009). Medline doi:10.1016/j.cell.2009.09.033
47. C. A. Hoeffer, H. Wong, P. Cain, J. Levenga, K. K. Cowansage, Y. Choi, C. Davy, N. Majmundar, D. R. McMillan, B. A. Rothermel, E. Klann, Regulator of calcineurin 1 modulates expression of innate anxiety and anxiogenic responses to selective serotonin reuptake inhibitor treatment. J. Neurosci. 33, 16930–16944 (2013). Medline doi:10.1523/JNEUROSCI.3513-12.2013