Animal Reproduction (AR)
Animal Reproduction (AR)
Original Article

Changes in expression of interferon-stimulated genes and ubiquitin activating enzyme E1-like in ovine thymus during early pregnancy

Leying Zhang; Zimo Zhao; Yujiao Wang; Ning Li; Nan Cao; Ling Yang

Downloads: 0
Views: 193


Abstract: As the main signal for the maternal recognition in ruminants, interferon-tau (IFNT) stimulates expression of interferon-stimulated genes (ISGs) in uterus and many extrauterine tissues. However, it is unclear that early pregnancy induces expression of signal transducer and activator of transcription 1 (STAT1), myxovirusresistance 1 (Mx1), interferon-gamma-inducible protein 10 (IP-10) and ubiquitin activating enzyme E1-like protein (UBE1L) in maternal thymus. In this study, ovine thymuses were sampled on day 16 of the estrous cycle and on days 13, 16 and 25 of gestation, and the expression of STAT1, Mx1, IP-10 and UBE1L was detected by real-time quantitative PCR, Western blot and immunohistochemistry. The results revealed that the expression of STAT1 and IP-10 reached peaks on day 16 of pregnancy, and expression of Mx1 was enhanced on day 25 of pregnancy, and STAT1 protein was located in the epithelial reticular cells, capillaries and thymic corpuscles. However, expression of UBE1L was declined during early pregnancy. In conclusion, early pregnancy influences expression of STAT1, Mx1, IP-10 and UBE1L in maternal thymus, which may participate in regulation of maternal immune tolerance during early pregnancy in sheep.


interferon-gamma-inducible protein 10, myxovirusresistance 1, sheep, signal transducer and activator of transcription 1, ubiquitin activating enzyme E1-like


Basavaraja R, Przygrodzka E, Pawlinski B, Gajewski Z, Kaczmarek MM, Meidan R. Interferon-tau promotes luteal endothelial cell survival and inhibits specific luteolytic genes in bovine corpus luteum. Reproduction. 2017;154(5):559-68. PMid:28808111.

Bazer FW, Thatcher WW. Chronicling the discovery of interferon tau. Reproduction. 2017;154(5):F11-20. PMid:28747540.

Binelli M, Subramaniam P, Diaz T, Johnson G, Hansen TR, Badinga L, Thatcher WW. Bovine interferon-tau stimulates the Janus kinase-signal transducer and activator of transcription pathway in bovine endometrial epithelial cells. Biol Reprod. 2001;64(2):654-65. PMid:11159370.

Bott RC, Ashley RL, Henkes LE, Antoniazzi AQ, Bruemmer JE, Niswender GD, Bazer FW, Spencer TE, Smirnova NP, Anthony RV, Hansen TR. Uterine vein infusion of interferon tau (IFNT) extends luteal life span in ewes. Biol Reprod. 2010;82(4):725-35. PMid:20042537.

Calder A, Hince M, Dudakov J, Chidgey A, Boyd R. Thymic involution: where endocrinology meets immunology. Neuroimmunomodulation. 2011;18(5):281-9. PMid:21952680.

Chambers SP, Clarke AG. Measurement of thymus weight, lumbar node weight and progesterone levels in syngeneically pregnant, allogeneically pregnant, and pseudopregnant mice. J Reprod Fertil. 1979;55(2):309-15. PMid:439064.

Choi Y, Johnson GA, Burghardt RC, Berghman LR, Joyce MM, Taylor KM, Stewart MD, Bazer FW, Spencer TE. Interferon regulatory factor-two restricts expression of interferon-stimulated genes to the endometrial stroma and glandular epithelium of the ovine uterus. Biol Reprod. 2001;65(4):1038-49. PMid:11566724.

Durfee LA, Huibregtse JM. Identification and validation of ISG15 target proteins. Subcell Biochem. 2010;54:228-37. PMid:21222286.

Ebner P, Versteeg GA, Ikeda F. Ubiquitin enzymes in the regulation of immune responses. Crit Rev Biochem Mol Biol. 2017;52(4):425-60. PMid:28524749.

Gao S, von der Malsburg A, Dick A, Faelber K, Schröder GF, Haller O, Kochs G, Daumke O. Structure of myxovirus resistance protein a reveals intra- and intermolecular domain interactions required for the antiviral function. Immunity. 2011;35(4):514-25. PMid:21962493.

Gattass CR, King LB, Luster AD, Ashwell JD. Constitutive expression of interferon gamma-inducible protein 10 in lymphoid organs and inducible expression in T cells and thymocytes. J Exp Med. 1994;179(4):1373-8. PMid:8145049.

Gifford CA, Assiri AM, Satterfield MC, Spencer TE, Ott TL. Receptor transporter protein 4 (RTP4) in endometrium, ovary, and peripheral blood leukocytes of pregnant and cyclic ewes. Biol Reprod. 2008;79(3):518-24. PMid:18495679.

Godkin JD, Bazer FW, Roberts RM. Ovine trophoblast protein 1, an early secreted blastocyst protein, binds specifically to uterine endometrium and affects protein synthesis. Endocrinology. 1984;114(1):120-30. PMid:6418522.

Haq IU, Han Y, Ali T, Wang Y, Gao H, Lin L, Wu Y, Wu S, Zeng S. Expression of interferon-stimulated gene ISG15 and ubiquitination enzymes is upregulated in peripheral blood monocyte during early pregnancy in dairy cattle. Reprod Biol. 2016;16(4):255-60. PMid:27802914.

Hubel P, Urban C, Bergant V, Schneider WM, Knauer B, Stukalov A, Scaturro P, Mann A, Brunotte L, Hoffmann HH, Schoggins JW, Schwemmle M, Mann M, Rice CM, Pichlmair A. A protein-interaction network of interferon-stimulated genes extends the innate immune system landscape. Nat Immunol. 2019;20(4):493-502. PMid:30833792.

Imakawa K, Anthony RV, Kazemi M, Marotti KR, Polites HG, Roberts RM. Interferon-like sequence of ovine trophoblast protein secreted by embryonic trophectoderm. Nature. 1987;330(6146):377-9. PMid:2446135.

Imakawa K, Imai M, Sakai A, Suzuki M, Nagaoka K, Sakai S, Lee SR, Chang KT, Echternkamp SE, Christenson RK. Regulation of conceptus adhesion by endometrial CXC chemokines during the implantation period in sheep. Mol Reprod Dev. 2006;73(7):850-8. PMid:16596627.

Johnson GA, Spencer TE, Hansen TR, Austin KJ, Burghardt RC, Bazer FW. Expression of the interferon tau inducible ubiquitin cross-reactive protein in the ovine uterus. Biol Reprod. 1999;61(1):312-8. PMid:10377064.

Kappler JW, Roehm N, Marrack P. T cell tolerance by clonal elimination in the thymus. Cell. 1987;49(2):273-80. PMid:3494522.

Kim KI, Yan M, Malakhova O, Luo JK, Shen MF, Zou W, de la Torre JC, Zhang DE. Ube1L and protein ISGylation are not essential for alpha/beta interferon signaling. Mol Cell Biol. 2006;26(2):472-9. PMid:16382139.

Kunii H, Koyama K, Ito T, Suzuki T, Balboula AZ, Shirozu T, Bai H, Nagano M, Kawahara M, Takahashi M. Hot topic: pregnancy-induced expression of interferon-stimulated genes in the cervical and vaginal mucosal membranes. J Dairy Sci. 2018;101(9):8396-400. PMid:29935833.

Li J, Roberts RM. Interferon-tau and interferon-alpha interact with the same receptors in bovine endometrium. Use of a readily iodinatable form of recombinant interferon-tau for binding studies. J Biol Chem. 1994;269(18):13544-50. PMid:8175789.

Lynch HE, Goldberg GL, Chidgey A, Van den Brink MR, Boyd R, Sempowski GD. Thymic involution and immune reconstitution. Trends Immunol. 2009;30(7):366-73. PMid:19540807.

Mauffré V, Grimard B, Eozenou C, Inghels S, Silva L, Giraud-Delville C, Capo D, Sandra O, Constant F. Interferon stimulated genes as peripheral diagnostic markers of early pregnancy in sheep: a critical assessment. Animal. 2016;10(11):1856-63. PMid:27150201.

Meyerholz MM, Mense K, Knaack H, Sandra O, Schmicke M. Pregnancy-induced ISG-15 and MX-1 gene expression is detected in the liver of holstein-friesian heifers during late peri-implantation period. Reprod Domest Anim. 2016;51(1):175-7. PMid:26549692.

Mirando MA, Short EC Jr, Geisert RD, Vallet JL, Bazer FW. Stimulation of 2′,5′-oligoadenylate synthetase activity in sheep endometrium during pregnancy, by intrauterine infusion of ovine trophoblast protein-1, and by intramuscular administration of recombinant bovine interferon-alpha I1. J Reprod Fertil. 1991;93(2):599-607. PMid:1787480.

Nagaoka K, Nojima H, Watanabe F, Chang KT, Christenson RK, Sakai S, Imakawa K. Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon gamma-inducible protein 10 kDa (IP-10), during early gestation. J Biol Chem. 2003;278(31):29048-56. PMid:12756249.

Oliveira JF, Henkes LE, Ashley RL, Purcell SH, Smirnova NP, Veeramachaneni DN, Anthony RV, Hansen TR. Expression of interferon (IFN)-stimulated genes in extrauterine tissues during early pregnancy in sheep is the consequence of endocrine IFN-tau release from the uterine vein. Endocrinology. 2008;149(3):1252-9. PMid:18063687.

Ott TL, Yin J, Wiley AA, Kim HT, Gerami-Naini B, Spencer TE, Bartol FF, Burghardt RC, Bazer FW. Effects of the estrous cycle and early pregnancy on uterine expression of Mx protein in sheep (Ovis aries). Biol Reprod. 1998;59(4):784-94. PMid:9746726.

Racicot K, Ott T. The myxovirus resistance protein, MX1, interacts with tubulin beta in uterine glandular epithelial cells. Am J Reprod Immunol. 2011;65(1):44-53. PMid:20584010.

Racicot K, Schmitt A, Ott T. The myxovirus-resistance protein, MX1, is a component of exosomes secreted by uterine epithelial cells. Am J Reprod Immunol. 2012;67(6):498-505. PMid:22574859.

Rempel LA, Francis BR, Austin KJ, Hansen TR. Isolation and sequence of an interferon-tau-inducible, pregnancy- and bovine interferon-stimulated gene product 15 (ISG15)-specific, bovine ubiquitin-activating E1-like (UBE1L) enzyme. Biol Reprod. 2005;72(2):365-72. PMid:15385418.

Ribatti D, Crivellato E, Vacca A. Miller’s seminal studies on the role of thymus in immunity. Clin Exp Immunol. 2006;144(3):371-5. PMid:16734604.

Romagnani P, Crescioli C. CXCL10: a candidate biomarker in transplantation. Clin Chim Acta. 2012;413(17-18):1364-73. PMid:22366165.

Romero JJ, Antoniazzi AQ, Smirnova NP, Webb BT, Yu F, Davis JS, Hansen TR. Pregnancy-associated genes contribute to antiluteolytic mechanisms in ovine corpus luteum. Physiol Genomics. 2013;45(22):1095-108. PMid:24046284.

Sakumoto R, Iga K, Hayashi KG, Fujii S, Kanahara H, Hosoe M, Furusawa T. Gene expression of CCL8 and CXCL10 in peripheral blood leukocytes during early pregnancy in cows. J Anim Sci Biotechnol. 2018;9(1):46. PMid:29946459.

Seif F, Khoshmirsafa M, Aazami H, Mohsenzadegan M, Sedighi G, Bahar M. The role of JAK-STAT signaling pathway and its regulators in the fate of T helper cells. Cell Commun Signal. 2017;15(1):23. PMid:28637459.

Sheikh AA, Hooda OK, Kalyan A, Kamboj A, Mohammed S, Alhussien M, Reddi S, Shimray PG, Rautela A, Pandita S, Kapila S, De S, Dang AK. Interferon-tau stimulated gene expression: a proxy to predict embryonic mortality in dairy cows. Theriogenology. 2018;120:61-7. PMid:30096617.

Shinomiya N, Tsuru S, Tsugita M, Katsura Y, Takemura T, Rokutanda M, Nomoto K. Thymic depletion in pregnancy: kinetics of thymocytes and immunologic capacities of the hosts. J Clin Lab Immunol. 1991;34(1):11-22. PMid:1667937.

Shirozu T, Sasaki K, Kawahara M, Yanagawa Y, Nagano M, Yamauchi N, Takahashi M. Expression dynamics of bovine MX genes in the endometrium and placenta during early to mid pregnancy. J Reprod Dev. 2016;62(1):29-35. PMid:26498202.

Spencer TE. Biological roles of uterine glands in pregnancy. Semin Reprod Med. 2014;32(5):346-57. PMid:24959816.

Vitorino Carvalho A, Eozenou C, Healey GD, Forde N, Reinaud P, Chebrout M, Gall L, Rodde N, Padilla AL, Delville CG, Leveugle M, Richard C, Sheldon IM, Lonergan P, Jolivet G, Sandra O. Analysis of STAT1 expression and biological activity reveals interferon-tau-dependent STAT1-regulated SOCS genes in the bovine endometrium. Reprod Fertil Dev. 2016;28(4):459-74. PMid:25116692.

Wang P, Peng X, Zhang J, Wang Z, Meng J, Cen B, Ji A, He S. LncRNA-135528 inhibits tumor progression by up-regulating CXCL10 through the JAK/STAT pathway. Apoptosis. 2018;23(11-12):651-66. PMid:30232656.

Williams Z. Inducing tolerance to pregnancy. N Engl J Med. 2012;367(12):1159-61. PMid:22992082.

Wong ML, Medrano JF. Real-time PCR for mRNA quantitation. Biotechniques. 2005;39(1):75-85. PMid:16060372.

Xing Y, Wang X, Jameson SC, Hogquist KA. Late stages of T cell maturation in the thymus involve NF-κB and tonic type I interferon signaling. Nat Immunol. 2016;17(5):565-73. PMid:27043411.

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. 2017;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. 2018;89(11):1540-8. PMid:30191656.

Yang L, Lv W, Liu Y, Chen K, Xue J, Wang Q, Wang B, Zhang L. Effect of early pregnancy on the expression of prostaglandin synthases in the ovine thymus. Theriogenology. 2019a;136:166-71. PMid:31265945.

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. 2019b;86(1):100-8. PMid:30411425.

Yin X, Cong X, Yan M, Zhang DE. Deficiency of a potential 3p21.3 tumor suppressor gene UBE1L (UBA7) does not accelerate lung cancer development in K-rasLA2 mice. Lung Cancer. 2009;63(2):194-200. PMid:18571763.

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. PMid:30149261.

Zhang L, Zhao Z, Mi H, Liu B, Wang B, Yang L. Modulation of helper T cytokines in thymus during early pregnancy in ewes. Animals. 2019;9(5):245. PMid:31100843.

5f0473c10e8825851ebb8295 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections