Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2022-0093
Animal Reproduction (AR)
ORIGINAL ARTICLE

Changes in expression levels of Nod-like receptors in the spleen of ewes

Jiaxuan Wu; Shengya Fang; Pengfei Feng; Chunjiang Cai; Leying Zhang; Ling Yang

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Abstract

Nucleotide-binding oligomerization domain receptors (NOD-like receptors, NLRs) have critical effects on interfaces of the immune and reproductive systems, and the spleen plays a key role in both innate and adaptive immune functions. It is hypothesized that NLR family participates in maternal splenic immune regulation during early pregnancy in sheep. In this study, maternal spleens were collected on day 16 of the estrous cycle, and days 13, 16 and 25 of gestation (n = 6 for each group) in ewes. Expression of NLR family, including NOD1, NOD2, class II transactivator (CIITA), NLR family apoptosis inhibitory protein (NAIP), nucleotide-binding oligomerization domain, Leucine rich repeat and Pyrin domain containing 1 (NLRP1), NLRP3 and NLRP7, was analyzed using quantitative real-time PCR, Western blot and immunohistochemistry analysis. The results revealed that expression levels of NOD1, NOD2, CIITA and NLRP3 were downregulated at days 13 and 16 of pregnancy, but expression of NLRP3 was increased at day 25 of pregnancy. In addition, expression values of NAIP and NLRP7 mRNA and proteins were improved at days 16 and 25 of pregnancy, and NLRP1 was peaked at days 13 and 16 of pregnancy in the maternal spleen. Furthermore, NOD2 and NLRP7 proteins were limited to the capsule, trabeculae and splenic cords. In summary, early pregnancy changes expression of NLR family in the maternal spleen, which may be related with the maternal splenic immunomodulation during early pregnancy in sheep.

Keywords

Nod-like receptor, pregnancy, sheep, spleen

References

Abadía-Molina F, Morón-Calvente V, Baird SD, Shamim F, Martín F, MacKenzie A. Neuronal apoptosis inhibitory protein (NAIP) localizes to the cytokinetic machinery during cell division. Sci Rep. 2017;7(1):39981. http://dx.doi.org/10.1038/srep39981. PMid:28059125.

Abi Nahed R, Elkhoury Mikhael M, Reynaud D, Collet C, Lemaitre N, Michy T, Hoffmann P, Sergent F, Marquette C, Murthi P, Raia-Barjat T, Alfaidy N, Benharouga M. Role of NLRP7 in normal and malignant trophoblast cells. Biomedicines. 2022;10(2):252. http://dx.doi.org/10.3390/biomedicines10020252. PMid:35203462.

Abrahams VM. Pattern recognition at the maternal-fetal interface. Immunol Invest. 2008;37(5):427-47. http://dx.doi.org/10.1080/08820130802191599. PMid:18716932.

Bai J, Zhang L, Zhao Z, Li N, Wang B, Yang L. Expression of melatonin receptors and CD4 in the ovine thymus, lymph node, spleen and liver during early pregnancy. Immunology. 2020;160(1):52-63. http://dx.doi.org/10.1111/imm.13180. PMid:32052861.

Cao N, Cao L, Gao M, Wang H, Zhang L, Yang L. Changes in mRNA and protein levels of gonadotropin releasing hormone and receptor in ovine thymus, lymph node, spleen, and liver during early pregnancy. Domest Anim Endocrinol. 2021;76:106607. http://dx.doi.org/10.1016/j.domaniend.2021.106607. PMid:33582417.

Carp H. Immunotherapy for recurrent pregnancy loss. Best Pract Res Clin Obstet Gynaecol. 2019;60:77-86. http://dx.doi.org/10.1016/j.bpobgyn.2019.07.005. PMid:31521575.

Carriere J, Dorfleutner A, Stehlik C. NLRP7: from inflammasome regulation to human disease. Immunology. 2021;163(4):363-76. http://dx.doi.org/10.1111/imm.13372. PMid:34021586.

Costa FRC, Leite JA, Rassi DM, da Silva JF, Elias-Oliveira J, Guimarães JB, Foss-Freitas MC, Câmara NOS, Pontillo A, Tostes RC, Silva JS, Carlos D. NLRP1 acts as a negative regulator of Th17 cell programming in mice and humans with autoimmune diabetes. Cell Rep. 2021;35(8):109176. http://dx.doi.org/10.1016/j.celrep.2021.109176. PMid:34038731.

Crane GM, Liu YC, Chadburn A. Spleen: development, anatomy and reactive lymphoid proliferations. Semin Diagn Pathol. 2021;38(2):112-24. http://dx.doi.org/10.1053/j.semdp.2020.06.003. PMid:32591155.

Devaiah BN, Singer DS. CIITA and its dual roles in MHC gene transcription. Front Immunol. 2013;4:476. http://dx.doi.org/10.3389/fimmu.2013.00476. PMid:24391648.

Diez E, Yaraghi Z, MacKenzie A, Gros P. The neuronal apoptosis inhibitory protein (Naip) is expressed in macrophages and is modulated after phagocytosis and during intracellular infection with Legionella pneumophila. J Immunol. 2000;164(3):1470-7. http://dx.doi.org/10.4049/jimmunol.164.3.1470. PMid:10640764.

Elliott EI, Sutterwala FS. Initiation and perpetuation of NLRP3 inflammasome activation and assembly. Immunol Rev. 2015;265(1):35-52. http://dx.doi.org/10.1111/imr.12286. PMid:25879282.

Feng P, Wu J, Ren Y, Zhang L, Cao J, Yang L. Early pregnancy regulates the expression of prolactin and its receptor in the thymus, the liver, the spleen and lymph nodes in sheep. Domest Anim Endocrinol. 2022;81:106731. http://dx.doi.org/10.1016/j.domaniend.2022.106731. PMid:35635981.

Fernández-García V, González-Ramos S, Avendaño-Ortiz J, Martín-Sanz P, Delgado C, Castrillo A, Boscá L. NOD1 splenic activation confers ferroptosis protection and reduces macrophage recruitment under pro-atherogenic conditions. Biomed Pharmacother. 2022;148:112769. http://dx.doi.org/10.1016/j.biopha.2022.112769. PMid:35247718.

Fernández-García V, González-Ramos S, Martín-Sanz P, García-Del Portillo F, Laparra JM, Boscá L. NOD1 in the interplay between microbiota and gastrointestinal immune adaptations. Pharmacol Res. 2021;171:105775. http://dx.doi.org/10.1016/j.phrs.2021.105775. PMid:34273489.

Figueiredo AS, Schumacher A. The T helper type 17/regulatory T cell paradigm in pregnancy. Immunology. 2016;148(1):13-21. http://dx.doi.org/10.1111/imm.12595. PMid:26855005.

Fröhlich C, Ehrhardt J, Krüger D, Trojnarska D, Zygmunt M, Muzzio DO. Pregnancy status alters IL-21-mediated effects on murine B lymphocytes. Reproduction. 2020;159(3):351-9. http://dx.doi.org/10.1530/REP-19-0407. PMid:31940277.

Genebrier S, Tarte K. The flawless immune tolerance of pregnancy. Joint Bone Spine. 2021;88(5):105205. http://dx.doi.org/10.1016/j.jbspin.2021.105205. PMid:33962032.

Hao S, Fang H, Fang S, Zhang T, Zhang L, Yang L. Changes in nuclear factor kappa B components expression in the ovine spleen during early pregnancy. J Anim Feed Sci. 2022;31(1):3-11. http://dx.doi.org/10.22358/jafs/146491/2022.

Huang JY, Yu PH, Li YC, Kuo PL. NLRP7 contributes to in vitro decidualization of endometrial stromal cells. Reprod Biol Endocrinol. 2017;15(1):66. http://dx.doi.org/10.1186/s12958-017-0286-x. PMid:28810880.

Ingram-Crooks J, Holcik M, Drmanic S, MacKenzie AE. Distinct expression of neuronal apoptosis inhibitory protein (NAIP) during murine development. Neuroreport. 2002;13(4):397-402. http://dx.doi.org/10.1097/00001756-200203250-00007. PMid:11930148.

Ka H, Hunt JS. Temporal and spatial patterns of expression of inhibitors of apoptosis in human placentas. Am J Pathol. 2003;163(2):413-22. http://dx.doi.org/10.1016/S0002-9440(10)63671-1. PMid:12875963.

King AE, Horne AW, Hombach-Klonisch S, Mason JI, Critchley HO. Differential expression and regulation of nuclear oligomerization domain proteins NOD1 and NOD2 in human endometrium: a potential role in innate immune protection and menstruation. Mol Hum Reprod. 2009;15(5):311-9. http://dx.doi.org/10.1093/molehr/gap020. PMid:19273470.

Lappas M. NOD1 and NOD2 regulate proinflammatory and prolabor mediators in human fetal membranes and myometrium via nuclear factor-kappa B. Biol Reprod. 2013;89(1):14. http://dx.doi.org/10.1095/biolreprod.113.110056. PMid:23740944.

León Machado JA, Steimle V. The MHC Class II Transactivator CIITA: Not (Quite) the Odd-One-Out Anymore among NLR Proteins. Int J Mol Sci. 2021;22(3):1074. http://dx.doi.org/10.3390/ijms22031074. PMid:33499042.

Li G, Tian X, Lv D, Zhang L, Zhang Z, Wang J, Yang M, Tao J, Ma T, Wu H, Ji P, Wu Y, Lian Z, Cui W, Liu G. NLRP7 is expressed in the ovine ovary and associated with in vitro pre-implantation embryo development. Reproduction. 2019a;158(5):415-27. http://dx.doi.org/10.1530/REP-19-0081. PMid:31505467.

Li N, Zhao Z, Bai J, Liu B, Mi H, Zhang L, Li G, Yang L. Characterization of the T help cytokines profile in ovine spleen during early pregnancy. J Appl Anim Res. 2019b;47(1):386-93. http://dx.doi.org/10.1080/09712119.2019.1634077.

Li M, Wu X, An P, Dang H, Liu Y, Liu R. Effects of resveratrol on autophagy and the expression of inflammasomes in a placental trophoblast oxidative stress model. Life Sci. 2020;256:117890. http://dx.doi.org/10.1016/j.lfs.2020.117890. PMid:32497634.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25(4):402-8. http://dx.doi.org/10.1006/meth.2001.1262. PMid:11846609.

Matias ML, Gomes VJ, Romao-Veiga M, Ribeiro VR, Nunes PR, Romagnoli GG, Peracoli JC, Peracoli MTS. Silibinin Downregulates the NF-κB Pathway and NLRP1/NLRP3 Inflammasomes in Monocytes from Pregnant Women with Preeclampsia. Molecules. 2019;24(8):1548. http://dx.doi.org/10.3390/molecules24081548. PMid:31010153.

Meihe L, Shan G, Minchao K, Xiaoling W, Peng A, Xili W, Jin Z, Huimin D. The ferroptosis-NLRP1 inflammasome: the vicious cycle of an adverse pregnancy. Front Cell Dev Biol. 2021;9:707959. http://dx.doi.org/10.3389/fcell.2021.707959. PMid:34490257.

Mulla MJ, Myrtolli K, Potter J, Boeras C, Kavathas PB, Sfakianaki AK, Tadesse S, Norwitz ER, Guller S, Abrahams VM. Uric acid induces trophoblast IL-1β production via the inflammasome: implications for the pathogenesis of preeclampsia. Am J Reprod Immunol. 2011;65(6):542-8. http://dx.doi.org/10.1111/j.1600-0897.2010.00960.x. PMid:21352397.

Nagatomo H, Akizawa H, Sada A, Kishi Y, Yamanaka K, Takuma T, Sasaki K, Yamauchi N, Yanagawa Y, Nagano M, Kono T, Takahashi M, Kawahara M. Comparing spatial expression dynamics of bovine blastocyst under three different procedures: in-vivo, in-vitro derived, and somatic cell nuclear transfer embryos. Jpn J Vet Res. 2015;63(4):159-71. PMid:26753242.

Napier RJ, Lee EJ, Vance EE, Snow PE, Samson KA, Dawson CE, Moran AE, Stenzel P, Davey MP, Sakaguchi S, Rosenzweig HL. Nod2 deficiency augments Th17 responses and exacerbates autoimmune arthritis. J Immunol. 2018;201(7):1889-98. http://dx.doi.org/10.4049/jimmunol.1700507. PMid:30150283.

Ott TL. Immunological detection of pregnancy: evidence for systemic immune modulation during early pregnancy in ruminants. Theriogenology. 2020;150:498-503. http://dx.doi.org/10.1016/j.theriogenology.2020.04.010. PMid:32331860.

Prescott D, Maisonneuve C, Yadav J, Rubino SJ, Girardin SE, Philpott DJ. NOD2 modulates immune tolerance via the GM-CSF-dependent generation of CD103+ dendritic cells. Proc Natl Acad Sci USA. 2020;117(20):10946-57. http://dx.doi.org/10.1073/pnas.1912866117. PMid:32350141.

Rakner JJ, Silva GB, Mundal SB, Thaning AJ, Elschot M, Ostrop J, Thomsen LCV, Bjørge L, Gierman LM, Iversen AC. Decidual and placental NOD1 is associated with inflammation in normal and preeclamptic pregnancies. Placenta. 2021;105:23-31. http://dx.doi.org/10.1016/j.placenta.2021.01.014. PMid:33529885.

Rocha CC, Silveira JC, Forde N, Binelli M, Pugliesi G. Conceptus-modulated innate immune function during early pregnancy in ruminants: a review. Anim Reprod. 2021;18(1):e20200048. http://dx.doi.org/10.1590/1984-3143-ar2020-0048. PMid:34122650.

Soczewski E, Grasso E, Gallino L, Hauk V, Fernández L, Gori S, Paparini D, Perez Leirós C, Ramhorst R. Immunoregulation of the decidualization program: focus on the endoplasmic reticulum stress. Reproduction. 2020;159(4):R203-11. http://dx.doi.org/10.1530/REP-19-0391. PMid:31990665.

Sundaram B, Kanneganti TD. Advances in Understanding Activation and Function of the NLRC4 Inflammasome. Int J Mol Sci. 2021;22(3):1048. http://dx.doi.org/10.3390/ijms22031048. PMid:33494299.

van Gorp H, Kuchmiy A, Van Hauwermeiren F, Lamkanfi M. NOD-like receptors interfacing the immune and reproductive systems. FEBS J. 2014;281(20):4568-82. http://dx.doi.org/10.1111/febs.13014. PMid:25154302.

Wang S, Li M, Sun F, Chen C, Ye J, Li D, Qian J, Du M. Altered frequency and function of spleen CTLA-4+Tim-3+ T cells are associated with miscarriage. Biol Reprod. 2021;104(2):410-7. http://dx.doi.org/10.1093/biolre/ioz076. PMid:31329823.

Wang Y, Han X, Zhang L, Cao N, Cao L, Yang L. Early pregnancy induces expression of STAT1, OAS1 and CXCL10 in ovine spleen. Animals (Basel). 2019;9(11):882. http://dx.doi.org/10.3390/ani9110882. PMid:31671580.

Wei J, Chen Q, James JL, Stone PR, Chamley LW. IL-1 beta but not the NALP3 inflammasome is an important determinant of endothelial cell responses to necrotic/dangerous trophoblastic debris. Placenta. 2015;36(12):1385-92. http://dx.doi.org/10.1016/j.placenta.2015.10.011. PMid:26515928.

Yang L, Guo R, Yao X, Yan J, Bai Y, Zhang L. Expression of progesterone receptor and progesterone-induced blocking factor in the spleen during early pregnancy in ewes. Livest Sci. 2018a;209:14-9. http://dx.doi.org/10.1016/j.livsci.2018.01.004.

Yang L, Liu B, Yan X, Zhang L, Gao F, Liu Z. Expression of ISG15 in bone marrow during early pregnancy in ewes. Kafkas Univ Vet Fak Derg. 2017a;23:767-72.

Yang L, Liu Y, Lv W, Wang P, Wang B, Xue J, Zhang L. Expression of interferon-stimulated gene 15-kDa protein, cyclooxygenase (COX) 1, COX-2, aldo-keto reductase family 1, member B1, and prostaglandin E synthase in the spleen during early pregnancy in sheep. Anim Sci J. 2018b;89(11):1540-8. http://dx.doi.org/10.1111/asj.13101. PMid:30191656.

Yang L, Wang L, Wu J, Wang H, Yang G, Zhang L. Changes in Expression of Complement Components in the Ovine Spleen during Early Pregnancy. Animals (Basel). 2021;11(11):3183. http://dx.doi.org/10.3390/ani11113183. PMid:34827915.

Yang L, Wang Q, Liu Y, Zhang L, Lv W, Liu B. Expression profiles of interferon-stimulated gene 15 and prostaglandin synthases in the ovine lymph nodes during early pregnancy. Mol Reprod Dev. 2019;86(1):100-8. http://dx.doi.org/10.1002/mrd.23085. PMid:30411425.

Yang L, Zang S, Bai Y, Yao X, Zhang L. Effect of early pregnancy on the expression of progesterone receptor and progesterone-induced blocking factor in ovine lymph node. Theriogenology. 2017b;93:78-83. http://dx.doi.org/10.1016/j.theriogenology.2017.01.042. PMid:28257871.

Yoo I, Kim D, Han J, Lee S, Hong M, Jeon BY, Kim JM, Ka H. Transcriptomic analysis of interferon-γ-regulated genes in endometrial explants and their possible role in regulating maternal endometrial immunity during the implantation period in pigs, a true epitheliochorial placentation species. Theriogenology. 2020;155:114-24. http://dx.doi.org/10.1016/j.theriogenology.2020.05.045. PMid:32659448.

Zhang P, Dixon M, Zucchelli M, Hambiliki F, Levkov L, Hovatta O, Kere J. Expression analysis of the NLRP gene family suggests a role in human preimplantation development. PLoS One. 2008;3(7):e2755. http://dx.doi.org/10.1371/journal.pone.0002755. PMid:18648497.

Zhang Y, Yang C, Fu S, Chen X, Zhang S, Li Y, Du M, Zhang J. Different expression of NOD2 in decidual stromal cells between normal and unexplained recurrent spontaneous abortion women during first trimester gestation. Int J Clin Exp Pathol. 2014a;7(12):8784-90. PMid:25674246.

Zhang YY, Chen H, Sun C, Wang HZ, Liu ML, Li YY, Nie XL, Du MR, Li DJ, Zhang JP. Expression and functional characterization of NOD2 in decidual stromal cells isolated during the first trimester of pregnancy. PLoS One. 2014b;9(6):e99612. http://dx.doi.org/10.1371/journal.pone.0099612. PMid:24932916.

Zhang L, Mi H, Yan JK, Yan XX, Yang L. Pregnancy-associated changes in expression of progesterone receptor and progesterone-induced blocking factor genes in bone marrow of ewes. Anim Reprod Sci. 2017;186:77-84. http://dx.doi.org/10.1016/j.anireprosci.2017.09.007. PMid:28947097.

Zhang L, Xue J, Wang Q, Lv W, Mi H, Liu Y, Yang L. Changes in expression of ISG15, progesterone receptor and progesterone-induced blocking factor in ovine thymus during early pregnancy. Theriogenology. 2018;121:153-9. http://dx.doi.org/10.1016/j.theriogenology.2018.08.018. PMid:30149261.

Zhang Y, Zhang Y, Li C, Fu S, Yang C, Song Y, Liu M, Wang Z, Liang P, Zhang J. NOD1 modulates decidual stromal cell function to maintain pregnancy in the early trimester. Cell Biochem Funct. 2019;37(7):464-73. http://dx.doi.org/10.1002/cbf.3417. PMid:31396989.

Zhang L, Cao L, Yang F, Han X, Wang Y, Cao N, Yang L. Relative abundance of interferon-stimulated genes STAT1, OAS1, CXCL10 and MX1 in ovine lymph nodes during early pregnancy. Anim Reprod Sci. 2020a;214:106285. http://dx.doi.org/10.1016/j.anireprosci.2020.106285. PMid:32087912.

Zhang L, Zhao Z, Wang Y, Li N, Cao N, Yang L. Changes in expression of interferon-stimulated genes and ubiquitin activating enzyme E1-like in ovine thymus during early pregnancy. Anim Reprod. 2020b;17(2):e20190134. http://dx.doi.org/10.1590/1984-3143-ar2019-0134. PMid:32714456.

Zhang L, Yang G, Zhang Q, Feng P, Gao M, Yang L. Early pregnancy affects expression of Toll-like receptor signaling members in ovine spleen. Anim Reprod. 2021;18(2):e20210009. http://dx.doi.org/10.1590/1984-3143-ar2021-0009. PMid:34394755.

Zhao L, Liang X, Ma Y, Li J, Liao S, Chen J, Wang C. AK002210 promotes the proliferation, migration and invasion of trophoblast cell through regulating miR-590/NAIP signal axis. Arch Biochem Biophys. 2020;688:108366. http://dx.doi.org/10.1016/j.abb.2020.108366. PMid:32387473.
 


Submitted date:
09/27/2022

Accepted date:
03/07/2023

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