TLR7 escapes X chromosome inactivation in immune cells
The X chromosome link to lupus
Nine of 10 individuals who develop systemic lupus erythematosus (SLE) are women. Furthermore, individuals with Klinefelter syndrome (47,XXY) also have increased incidence of SLE, suggesting that X chromosome dosage could be an important risk factor in SLE. Using sensitive quantification methods, Souyris et al. demonstrate that Toll-like receptor 7 (TLR7) that is encoded from the X chromosome escapes X inactivation in B cells and myeloid cells in females and Klinefelter individuals. TLR7 binds single-stranded RNA and activates type I interferon signaling, a pathway that is also activated in SLE patients. On the basis of this, the authors propose that biallelic expression of TLR7 contributes to greater SLE risk in individuals with two X chromosomes.
Abstract
Toll-like receptor 7 (TLR7) is critical to the induction of antiviral immunity, but TLR7 dosage is also a key pathogenic factor in systemic lupus erythematosus (SLE), an autoimmune disease with strong female bias. SLE prevalence is also elevated in individuals with Klinefelter syndrome, who carry one or more supernumerary X chromosomes, suggesting that the X chromosome complement contributes to SLE susceptibility. TLR7 is encoded by an X chromosome locus, and we examined here whether the TLR7 gene evades silencing by X chromosome inactivation in immune cells from women and Klinefelter syndrome males. Single-cell analyses of TLR7 allelic expression demonstrated that substantial fractions of primary B lymphocytes, monocytes, and plasmacytoid dendritic cells not only in women but also in Klinefelter syndrome males express TLR7 on both X chromosomes. Biallelic B lymphocytes from women displayed greater TLR7 transcriptional expression than the monoallelic cells, correlated with higher TLR7 protein expression in female than in male leukocyte populations. Biallelic B cells were preferentially enriched during the TLR7-driven proliferation of CD27+ plasma cells. In addition, biallelic cells showed a greater than twofold increase over monoallelic cells in the propensity to immunoglobulin G class switch during the TLR7-driven, T cell–dependent differentiation of naive B lymphocytes into immunoglobulin-secreting cells. TLR7 escape from X inactivation endows the B cell compartment with added responsiveness to TLR7 ligands. This finding supports the hypothesis that enhanced TLR7 expression owing to biallelism contributes to the higher risk of developing SLE and other autoimmune disorders in women and in men with Klinefelter syndrome.
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Supplementary Material
Summary
Materials and Methods
Fig. S1. Determination of TLR7 monoallelic or biallelic expression at the single-cell level.
Fig. S2. In situ visualization of monoallelic and biallelic TLR7 transcripts at the single-cell level.
Fig. S3. Relationship between transcript quantitation and TLR7 monoallelic or biallelic expression.
Fig. S4. Specific Western blot immunodetection of TLR7.
Fig. S5. Unstable TLR7 allelic expression in EBV-transformed B cells.
Table S1. PCR primer pairs used in the allele-of-origin assay.
Table S2. Primary antibodies.
Table S3. PCR primer pairs used in the preparation of the TLR7 RNA FISH probe.
Table S4. PCR primer pairs used in the preparation of the XIST RNA FISH probe.
Table S5. PCR primer pairs used in the preparation of the SLC25A6 RNA FISH probe.
Table S6. 2 × 2 contingency tables of TLR7 biallelic and monoallelic cell counts in IgG+ and IgG− B cells.
Movie S1. RNA FISH visualization of TLR7 escape from X inactivation in a memory B lymphocyte.
Resources
REFERENCES AND NOTES
1
L. B. Barreiro, M. Ben-Ali, H. Quach, G. Laval, E. Patin, J. K. Pickrell, C. Bouchier, M. Tichit, O. Neyrolles, B. Gicquel, J. R. Kidd, K. K. Kidd, A. Alcaïs, J. Ragimbeau, S. Pellegrini, L. Abel, J.-L. Casanova, L. Quintana-Murci, Evolutionary dynamics of human Toll-like receptors and their different contributions to host defense. PLOS Genet. 5, e1000562 (2009).
2
C. Petes, N. Odoardi, K. Gee, The Toll for trafficking: Toll-like receptor 7 delivery to the endosome. Front. Immunol. 8, 1075 (2017).
3
M. Swiecki, M. Colonna, The multifaceted biology of plasmacytoid dendritic cells. Nat. Rev. Immunol. 15, 471–485 (2015).
4
G. Kassiotis, J. P. Stoye, Immune responses to endogenous retroelements: Taking the bad with the good. Nat. Rev. Immunol. 16, 207–219 (2016).
5
K. A. Kirou, C. Lee, S. George, K. Louca, M. G. Peterson, M. K. Crow, Activation of the interferon-α pathway identifies a subgroup of systemic lupus erythematosus patients with distinct serologic features and active disease. Arthritis Rheum. 52, 1491–1503 (2005).
6
J. A. Deane, P. Pisitkun, R. S. Barrett, L. Feigenbaum, T. Town, J. M. Ward, R. A. Flavell, S. Bolland, Control of Toll-like receptor 7 expression is essential to restrict autoimmunity and dendritic cell proliferation. Immunity 27, 801–810 (2007).
7
P. Pisitkun, J. A. Deane, M. J. Difilippantonio, T. Tarasenko, A. B. Satterthwaite, S. Bolland, Autoreactive B cell responses to RNA-related antigens due to TLR7 gene duplication. Science 312, 1669–1672 (2006).
8
S. Subramanian, K. Tus, Q.-Z. Li, A. Wang, X.-H. Tian, J. Zhou, C. Liang, G. Bartov, L. D. McDaniel, X. J. Zhou, R. A. Schultz, E. K. Wakeland, A Tlr7 translocation accelerates systemic autoimmunity in murine lupus. Proc. Natl. Acad. Sci. U.S.A. 103, 9970–9975 (2006).
9
S. R. Christensen, J. Shupe, K. Nickerson, M. Kashgarian, R. A. Flavell, M. J. Shlomchik, Toll-like receptor 7 and TLR9 dictate autoantibody specificity and have opposing inflammatory and regulatory roles in a murine model of lupus. Immunity 25, 417–428 (2006).
10
S. W. Jackson, N. E. Scharping, N. S. Kolhatkar, S. Khim, M. A. Schwartz, Q.-Z. Li, K. L. Hudkins, C. E. Alpers, D. Liggitt, D. J. Rawlings, Opposing impact of B cell–intrinsic TLR7 and TLR9 signals on autoantibody repertoire and systemic inflammation. J. Immunol. 192, 4525–4532 (2014).
11
C. Soni, E. B. Wong, P. P. Domeier, T. N. Khan, T. Satoh, S. Akira, Z. S. Rahman, B cell–intrinsic TLR7 signaling is essential for the development of spontaneous germinal centers. J. Immunol. 193, 4400–4414 (2014).
12
A. A. Margery-Muir, C. Bundell, D. Nelson, D. M. Groth, J. D. Wetherall, Gender balance in patients with systemic lupus erythematosus. Autoimmun. Rev. 16, 258–268 (2017).
13
R. H. Scofield, G. R. Bruner, B. Namjou, R. P. Kimberly, R. Ramsey-Goldman, M. Petri, J. D. Reveille, G. S. Alarcón, L. M. Vilá, J. Reid, B. Harris, S. Li, J. A. Kelly, J. B. Harley, Klinefelter’s syndrome (47,XXY) in male systemic lupus erythematosus patients: Support for the notion of a gene-dose effect from the X chromosome. Arthritis Rheum. 58, 2511–2517 (2008).
14
O. O. Seminog, A. B. Seminog, D. Yeates, M. J. Goldacre, Associations between Klinefelter’s syndrome and autoimmune diseases: English national record linkage studies. Autoimmunity 48, 125–128 (2015).
15
M. F. Lyon, Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature 190, 372–373 (1961).
16
A. Sahakyan, K. Plath, C. Rougeulle, Regulation of X-chromosome dosage compensation in human: Mechanisms and model systems. Philos. Trans. R. Soc. Lond. B Biol. Sci. 372, 20160363 (2017).
17
L. Carrel, H. F. Willard, X-inactivation profile reveals extensive variability in X-linked gene expression in females. Nature 434, 400–404 (2005).
18
A. M. Cotton, B. Ge, N. Light, V. Adoue, T. Pastinen, C. J. Brown, Analysis of expressed SNPs identifies variable extents of expression from the human inactive X chromosome. Genome Biol. 14, R122 (2013).
19
L. Carrel, C. J. Brown, When the Lyon(ized chromosome) roars: Ongoing expression from an inactive X chromosome. Philos. Trans. R. Soc. Lond. B Biol. Sci. 372, 20160355 (2017).
20
T. Tukiainen, A. C. Villani, A. Yen, M. A. Rivas, J. L. Marshall, R. Satija, M. Aguirre, L. Gauthier, M. Fleharty, A. Kirby, B. B. Cummings, S. E. Castel, K. J. Karczewski, F. Aguet, A. ByrnesGTEx Consortium, T. Lappalainen, A. Regev, K. G. Ardlie, N. Hacohen, D. G. MacArthur, Landscape of X chromosome inactivation across human tissues. Nature 550, 244–248 (2017).
21
C. He, J. Holme, J. Anthony, SNP genotyping: The KASP assay, in Crop Breeding: Methods and Protocols, D. Fleury, R. Whitford, Eds. (Humana Press, 2014), pp. 75–86.
22
J. Chaumeil, S. Augui, J. C. Chow, E. Heard, Combined immunofluorescence, RNA fluorescent in situ hybridization, and DNA fluorescent in situ hybridization to study chromatin changes, transcriptional activity, nuclear organization, and X-chromosome inactivation, in The Nucleus, R. Hancock, Ed. (Humana Press, 2008), vol. 1, pp. 297–308.
23
C. M. Clemson, J. A. McNeil, H. F. Willard, J. B. Lawrence, XIST RNA paints the inactive X chromosome at interphase: Evidence for a novel RNA involved in nuclear/chromosome structure. J. Cell Biol. 132, 259–275 (1996).
24
B. Reinius, R. Sandberg, Random monoallelic expression of autosomal genes: Stochastic transcription and allele-level regulation. Nat. Rev. Genet. 16, 653–664 (2015).
25
B. T. Lahn, D. C. Page, Functional coherence of the human Y chromosome. Science 278, 675–680 (1997).
26
N. Simchoni, C. Cunningham-Rundles, TLR7- and TLR9-responsive human B cells share phenotypic and genetic characteristics. J. Immunol. 194, 3035–3044 (2015).
27
I. B. Bekeredjian-Ding, M. Wagner, V. Hornung, T. Giese, M. Schnurr, S. Endres, G. Hartmann, Plasmacytoid dendritic cells control TLR7 sensitivity of naive B cells via type I IFN. J. Immunol. 174, 4043–4050 (2005).
28
D. T. Avery, J. I. Ellyard, F. Mackay, L. M. Corcoran, P. D. Hodgkin, S. G. Tangye, Increased expression of CD27 on activated human memory B cells correlates with their commitment to the plasma cell lineage. J. Immunol. 174, 4034–4042 (2005).
29
R. Fukui, S.-i. Saitoh, A. Kanno, M. Onji, T. Shibata, A. Ito, M. Onji, M. Matsumoto, S. Akira, N. Yoshida, K. Miyake, Unc93B1 restricts systemic lethal inflammation by orchestrating Toll-like receptor 7 and 9 trafficking. Immunity 35, 69–81 (2011).
30
R. Fukui, S.-i. Saitoh, F. Matsumoto, H. Kozuka-Hata, M. Oyama, K. Tabeta, B. Beutler, K. Miyake, Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells toward DNA- but against RNA-sensing. J. Exp. Med. 206, 1339–1350 (2009).
31
C. Pasare, R. Medzhitov, Control of B-cell responses by Toll-like receptors. Nature 438, 364–368 (2005).
32
C. R. Ruprecht, A. Lanzavecchia, Toll-like receptor stimulation as a third signal required for activation of human naive B cells. Eur. J. Immunol. 36, 810–816 (2006).
33
M. C. Glaum, S. Narula, D. Song, Y. Zheng, A. L. Anderson, C. H. Pletcher, A. I. Levinson, Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production. J. Allergy Clin. Immunol. 123, 224–230.e224 (2009).
34
P. Webb, C. Bain, S. Pirozzo, in Essential Epidemiology (Cambridge University Press, ed. 2, 2005), pp. 106–111.
35
J. Zhang, K. F. Yu, What’s the relative risk? A method of correcting the odds ratio in cohort studies of common outcomes. JAMA 280, 1690–1691 (1998).
36
J. Wang, C. M. Syrett, M. C. Kramer, A. Basu, M. L. Atchison, M. C. Anguera, Unusual maintenance of X chromosome inactivation predisposes female lymphocytes for increased expression from the inactive X. Proc. Natl. Acad. Sci. U.S.A. 113, E2029–2038 (2016).
37
Y. Zhang, A. Castillo-Morales, M. Jiang, Y. Zhu, L. Hu, A. O. Urrutia, X. Kong, L. D. Hurst, Genes that escape X-inactivation in humans have high intraspecific variability in expression, are associated with mental impairment but are not slow evolving. Mol. Biol. Evol. 30, 2588–2601 (2013).
38
A. Boneparth, W. Huang, R. Bethunaickan, M. Woods, R. Sahu, S. Arora, M. Akerman, M. Lesser, A. Davidson, TLR7 influences germinal center selection in murine SLE. PLOS ONE 10, e0119925 (2015).
39
N. V. Giltiay, C. P. Chappell, X. Sun, N. Kolhatkar, T. H. Teal, A. E. Wiedeman, J. Kim, L. Tanaka, M. B. Buechler, J. A. Hamerman, T. Imanishi-Kari, E. A. Clark, K. B. Elkon, Overexpression of TLR7 promotes cell-intrinsic expansion and autoantibody production by transitional T1 B cells. J. Exp. Med. 210, 2773–2789 (2013).
40
I. Moisini, W. Huang, R. Bethunaickan, R. Sahu, P.-G. Ricketts, M. Akerman, T. Marion, M. Lesser, A. Davidson, The Yaa locus and IFN-α fine-tune germinal center B cell selection in murine systemic lupus erythematosus. J. Immunol. 189, 4305–4312 (2012).
41
D. J. Birmingham, J. E. Bitter, E. G. Ndukwe, S. Dials, T. R. Gullo, S. Conroy, H. N. Nagaraja, B. H. Rovin, L. A. Hebert, Relationship of circulating anti-C3b and anti-C1q IgG to lupus nephritis and its flare. Clin. J. Am. Soc. Nephrol. 11, 47–53 (2016).
42
X.-X. Chen, Y.-Q. Chen, S. Ye, Measuring decreased serum IgG sialylation: A novel clinical biomarker of lupus. Lupus 24, 948–954 (2015).
43
K. D. Hansen, S. Sabunciyan, B. Langmead, N. Nagy, R. Curley, G. Klein, E. Klein, D. Salamon, A. P. Feinberg, Large-scale hypomethylated blocks associated with Epstein-Barr virus–induced B-cell immortalization. Genome Res. 24, 177–184 (2014).
44
M. Dominguez-Villar, A.-S. Gautron, M. de Marcken, M. J. Keller, D. A. Hafler, TLR7 induces anergy in human CD4+ T cells. Nat. Immunol. 16, 118–128 (2015).
45
C. Jacquemin, N. Schmitt, C. Contin-Bordes, Y. Liu, P. Narayanan, J. Seneschal, T. Maurouard, D. Dougall, E. S. Davizon, H. Dumortier, I. Douchet, L. Raffray, C. Richez, E. Lazaro, P. Duffau, M.-E. Truchetet, L. Khoryati, P. Mercié, L. Couzi, P. Merville, T. Schaeverbeke, J.-F. Viallard, J.-L. Pellegrin, J.-F. Moreau, S. Muller, S. Zurawski, R. L. Coffman, V. Pascual, H. Ueno, P. Blanco, OX40 ligand contributes to human lupus pathogenesis by promoting T follicular helper response. Immunity 42, 1159–1170 (2015).
46
G. S. Garcia-Romo, S. Caielli, B. Vega, J. Connolly, F. Allantaz, Z. Xu, M. Punaro, J. Baisch, C. Guiducci, R. L. Coffman, F. J. Barrat, J. Banchereau, V. Pascual, Netting neutrophils are major inducers of type I IFN production in pediatric systemic lupus erythematosus. Sci. Transl. Med. 3, 73ra20 (2011).
47
S. Laffont, C. Seillet, J.-C. Guery, Estrogen receptor-dependent regulation of dendritic cell development and function. Front. Immunol. 8, 108 (2017).
48
S. Laffont, N. Rouquié, P. Azar, C. Seillet, J. Plumas, C. Aspord, J.-C. Guery, X-Chromosome complement and estrogen receptor signaling independently contribute to the enhanced TLR7-mediated IFN-α production of plasmacytoid dendritic cells from women. J. Immunol. 193, 5444–5452 (2014).
49
C. Ritz, F. Baty, J. C. Streibig, D. Gerhard, Dose-response analysis using R. PLOS ONE 10, e0146021 (2015).
50
J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, Fiji: An open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
51
D. Liu, R. Y. Liu, M. Xie, Exact meta-analysis approach for discrete data and its application to 2 × 2 tables with rare events. J. Am. Stat. Assoc. 109, 1450–1465 (2014).
52
D. Smeets, Y. Markaki, V. J. Schmid, F. Kraus, A. Tattermusch, A. Cerase, M. Sterr, S. Fiedler, J. Demmerle, J. Popken, H. Leonhardt, N. Brockdorff, T. Cremer, L. Schermelleh, M. Cremer, Three-dimensional super-resolution microscopy of the inactive X chromosome territory reveals a collapse of its active nuclear compartment harboring distinct Xist RNA foci. Epigenetics Chromatin 7, 8 (2014).
53
L. Chaperot, N. Bendriss, O. Manches, R. Gressin, M. Maynadie, F. Trimoreau, H. Orfeuvre, B. Corront, J. Feuillard, J.-J. Sotto, J.-C. Bensa, F. Brière, J. Plumas, M.-C. Jacob, Identification of a leukemic counterpart of the plasmacytoid dendritic cells. Blood 97, 3210–3217 (2001).
54
J. Di Domizio, A. Blum, M. Gallagher-Gambarelli, J. P. Molens, L. Chaperot, J. Plumas, TLR7 stimulation in human plasmacytoid dendritic cells leads to the induction of early IFN-inducible genes in the absence of type I IFN. Blood 114, 1794–1802 (2009).
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Published In
Science Immunology
Volume 3 | Issue 19
January 2018
January 2018
Copyright
Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.
This is an article distributed under the terms of the Science Journals Default License.
Submission history
Received: 6 September 2017
Accepted: 4 December 2017
Acknowledgments
We thank M. Cremer for the gift of a XIST probe and L. Dupré for EBV infectious supernatants. We also thank J. Plumas for the gift of the GEN2.2 and GENshTLR7 cell lines. We are grateful to the staff of core laboratories for excellent technical support, especially A.-L. Iscache, A. Gendras, F. L’Faqihi, P.-E. Paulet, and R. Romieu-Mourez [Centre de Physiopathologie Toulouse-Purpan (CPTP)]; F. Martins (GeT TQ); and E. Lhuillier (GeT Purpan). Funding: J.C. is supported by the Foundation for Cancer Research (ARC), France and the ATIP-Avenir starting grant program (INSERM/CNRS/Plan cancer; France). Work at CPTP was supported by grants from ARC, Arthritis Fondation Courtin, Fondation pour la Recherche Médicale (DEQ2000329169), Conseil Régional Midi-Pyrénées, and the French National Agency for Research on AIDS and Viral Hepatitis and by studentships to M.S. and P.A. from Fondation pour la Recherche Médicale (FDT20170437183) and SIDACTION, respectively. Author contributions: M.S., P.A., J.E.M., and J.-C.G. conceived and designed the laboratory investigation. J.-C.G., S.G., and C.P. conceived and designed the study of KS patients. M.S., P.A., J.E.M., and J.-C.G. designed and performed the experiments with help from C.C., D.D., A.C., and J.C. M.S., C.C., D.D., J.E.M., and J.-C.G. analyzed the data. M.S., J.E.M., and J.-C.G. wrote the manuscript with contributions from co-authors. Competing interests: The authors declare that they have no competing interests. Data and materials availability: Plasmids and cell lines generated in this work can be provided upon request, subject to a material transfer agreement.
Authors
Funding Information
Arthritis Foundation: award349390
Association pour la Recherche sur le Cancer: award349391
Association pour la Recherche sur le Cancer: award349392
Fondation pour la Recherche Médicale: award349393, DEQ2000329169
Inserm/CNRS/Plan cancer, ATIP-Avenir starting grant program: award349396
Conseil Régional Midi-Pyrénées: award349395
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