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

Differential expression of T helper cytokines in the liver during early pregnancy in sheep

Ling Yang, Jiachen Bai, Zimo Zhao, Ning Li, Yujiao Wang, Leying Zhang

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Liver plays important roles in the innate and adaptive immunity, and contributes to the maternal immune adjustments during pregnancy in mice and rats. T helper 1 (Th1) and Th2 cytokines are related to immune response. However, expression of Th1 and Th2 cytokines in maternal livers is unclear during early pregnancy in sheep. In this study, livers were collected on day 16 of the estrous cycle and on days 13, 16 and 25 of pregnancy (n = 6 for each group) in ewes, and qRT-PCR, western blot and immunohistochemistry were used to analyze the expression of Th1 and Th2 cytokines in the livers. Our results showed that interferon-gamma (IFN-γ), interleukin (IL)-2, IL-4, IL-6 and IL-10 were downregulated, and IL-5 was upregulated in the livers during early pregnancy. Furthermore, there was no effect for early pregnancy on expression of TNF-β in the livers, and the IFN-γ protein was limited to the endothelial cells of the proper hepatic arteries and portal veins. In conclusion, early pregnancy exerted its effect on the liver to regulate the Th cytokines expression, but there was no evident shift from Th1 to Th2 cytokines, which may be necessary for the maternal hepatic immune adjustments during early pregnancy in sheep.


liver, pregnancy, T helper cytokine, sheep.


Bauer J, Namineni S, Reisinger F, Zöller J, Yuan D, Heikenwälder M. 2012. Lymphotoxin, NF-ĸB, and cancer: the dark side of cytokines. Dig Dis, 30:453-468.

Beltman ME, Forde N, Lonergan P, Crowe MA. 2013. Altered endometrial immune gene expression in beef heifers with retarded embryos. Reprod Fertil Dev, 25:966-970.

Bilotas MA, Olivares CN, Ricci AG, Baston JI, Bengochea TS, Meresman GF, Barañao RI. 2015. Interplay between Endometriosis and Pregnancy in a Mouse Model. PLoS One, 10:e0124900.

Blitek A, Morawska E, Ziecik AJ. 2012. Regulation of expression and role of leukemia inhibitory factor and interleukin-6 in the uterus of early pregnant pigs. Theriogenology, 78:951-964.

Bustamante JJ, Copple BL, Soares MJ, Dai G. 2010. Gene profiling of maternal hepatic adaptations to pregnancy. Liver Int, 30:406-415.

Chen L, Grabowski KA, Xin JP, Coleman J, Huang Z, Espiritu B, Alkan S, Xie HB, Zhu Y, White FA, Clancy J Jr, Huang H. 2004. IL-4 induces differentiation and expansion of Th2 cytokine-producing eosinophils. J Immunol, 172:2059-2066.

Cheng SB, Sharma S. 2014. Interleukin-10: a pleiotropic regulator in pregnancy. Am J Reprod Immunol, 73:487-500.

Chen Y, Lv W, Jia J, Wang J, Yang J. 2016. Evaluation of serum concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-2, IL-10, and nitric oxide (NO) during the estrous cycle, early pregnancy and abortion in goats. Anim Reprod Sci, 174:73-79.

Dai G, Bustamante JJ, Zou Y, Myronovych A, Bao Q, Kumar S, Soares MJ. 2011. Maternal hepatic growth response to pregnancy in the mouse. Exp Biol Med, 236:1322-3132.

de Melo JO, Soto SF, Katayama IA, Wenceslau CF, Pires AG, Veras MM, Furukawa LN, de Castro I, Saldiva PH, Heimann JC. 2015. Inhalation of fine particulate matter during pregnancy increased IL-4 cytokine levels in the fetal portion of the placenta. Toxicol Lett, 232:475-480.

De M, Sanford TR, Wood GW. 1992. Interleukin-1, interleukin-6, and tumor necrosis factor alpha are produced in the mouse uterus during the estrous cycle and are induced by estrogen and progesterone. Dev Biol, 151:297-305.

Francisco NM, Hsu NJ, Keeton R, Randall P, Sebesho B, Allie N, Govender D, Quesniaux V, Ryffel B, Kellaway L, Jacobs M. 2015. TNF-dependent regulation and activation of innate immune cells are essential for host protection against cerebral tuberculosis. J Neuroinflammation, 12:125.

Franczak A, Wojciechowicz B, Zmijewska A, Kolakowska J, Kotwica G. 2013. The effect of interleukin 1β and interleukin 6 on estradiol-17β secretion in the endometrium of pig during early pregnancy and the estrous cycle. Theriogenology, 80:90-98.

Frevert U, Engelmann S, Zougbédé S, Stange J, Ng B, Matuschewski K, Liebes L, Yee H. 2005. Intravital observation of Plasmodium berghei sporozoite infection of the liver. PLOS Biol, 3:e192.

Godkin JD, Bazer FW, Moffatt J, Sessions F, Roberts RM. 1982. Purification and properties of a major, low molecular weight protein released by the trophoblast of sheep blastocysts at day 13-21. J Reprod Fertil, 65:141-150.

Han AR, Lee SK. 2018. Immune modulation of i.v. immunoglobulin in women with reproductive failure. Reprod Med Biol, 17:115-124.

Hansen PJ. 2011. The immunology of early pregnancy in farm animals. Reprod Domest Anim, 46:18-30.

Harmon QE, Engel SM, Wu MC, Moran, TM, Luo J, Stuebe AM, Avery CL, Olshan AF. 2014. Polymorphisms in inflammatory genes are associated with term small for gestational age and preeclampsia. Am J Reprod Immunol, 71:472-484.

Jonsson Y, Matthiesen L, Berg G, Ernerudh J, Nieminen K, Ekerfelt C. 2005. Indications of an altered immune balance in preeclampsia: a decrease in in vitro secretion of IL-5 and IL-10 from blood mononuclear cells and in blood basophil counts compared with normal pregnancy. J Reprod Immunol, 66:69-84.

Kandil D, Leiman G, Allegretta M, Trotman W, Pantanowitz L, Goulart R, Evans M. 2007. Glypican-3 immunocytochemistry in liver fine-needle aspirates: a novel stain to assist in the differentiation of benign and malignant liver lesions. Cancer, 111:316-322.


Kmieć Z. 2001. Cooperation of liver cells in health and disease. Adv Anat Embryol Cell Biol, 161:1-151.

Liao W, Lin JX, Leonard WJ. 2011. IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Opin

Immunol, 23:598-604.

Liao W, Lin JX, Leonard WJ. 2013. Interleukin-2 at the crossroads of effector responses, tolerance, and immunotherapy. Immunity, 38:13-25.

Lin YL, Liang YC, Chiang BL. 2007. Placental growth factor down-regulates type 1 T helper immune response by modulating the function of dendritic cells. J Leukoc Biol, 82:1473-1480.

Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25:402-408.

Maciel GS, Uscategui RR, de Almeida VT, Oliveira ME, Feliciano MA, Vicente WR. 2014. Quantity of IL-2, IL-4, IL-10, INF-γ, TNF-α and KC-like cytokines in serum of bitches with pyometra in different stages of oestrous cycle and pregnancy. Reprod Domest Anim, 49:701-704

Martínez FF, Knubel CP, Sánchez MC, Cervi L, Motrán CC. 2012. Pregnancy-specific glycoprotein 1a activates dendritic cells to provide signals for Th17-, Th2-, and Treg-cell polarization. Eur J Immunol, 42:1573-1584.

Mcnatty KP, Revefeim KJ, Young A. 1973. Peripheral plasma progesterone concentrations in sheep during the oestrous cycle. J Endocrinol, 58:219-225.

Mi Y, Lin A, Fiete D, Steirer L, Baenziger JU. 2014. Modulation of mannose and asialoglycoprotein receptor expression determines glycoprotein hormone half-life at critical points in the reproductive cycle. J Biol Chem, 289:12157-12167.

Molvarec A, Szarka A, Lázár L, Rigó J Jr. 2013. PP028. Serum cytokine profile in relation to the clinical features and laboratory parameters in women with preeclampsia. Pregnancy Hypertension, 3:77.

Nakagome K, Okunishi K, Imamura M, Harada H, Matsumoto T, Tanaka R, Miyazaki J, Yamamoto K, Dohi M. 2009. IFN-gamma attenuates antigen-induced overall immune response in the airway as a Th1-type immune regulatory cytokine. J Immunol, 183:209-220.

Ott TL, Gifford CA. 2010. Effects of early conceptus signals on circulating immune cells: lessons from domestic ruminants. Am J Reprod Immunol, 64:245-254.

Ozbilgin K, Turan A, Kahraman B, Atay C, Vatansever S, Uluer ET, Ozçakir T. 2015. Distribution of furin, TNF-α, and TGF-β2 in the endometrium of missed abortion and voluntary first trimester termination cases. Anal Quant Cytopathol Histpathol, 37:123-133.


Parker GA, Picut CA. 2005. Liver immunobiology. Toxicol Pathol, 33:52-62.

Prins JR, Gomez-Lopez N, Robertson SA. 2012. Interleukin-6 in pregnancy and gestational disorders. J Reprod Immunol, 95:1-14.

Raghupathy R, Makhseed M, Azizieh F, Omu A, Gupta M, Farhat R. 2000. Cytokine production by maternal lymphocytes during normal human pregnancy and in unexplained recurrent spontaneous abortion. Hum Reprod, 15:713-718.

Roberts CT, White CA, Wiemer NG, Ramsay A, Roberston SA. 2003. Altered placental development in interleukin-10 null mutant mice. Placenta, 17:S94-S99.

Ruddle NH. 2014. Lymphotoxin and TNF: how it all began-a tribute to the travelers. Cytokine Growth Factor Rev, 25:83-89.

Saito S. 2000. Cytokine network at the feto-maternal interface. J Reprod Immunol, 47:87-103.

Schroder K, Hertzog PJ, Ravasi T, Hume DA. 2004. Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol, 75:163-189.

Sokol CL, Barton GM, Farr AG, Medzhitov R. 2008. A mechanism for the initiation of allergen-induced T helper type 2 responses. Nat Immunol, 9:310-318.

Ting JP, Painter A, Zeleznik-Le NJ, MacDonald G, Moore TM, Brown A, Schwartz BD. 1994. YB-1 DNA-binding protein represses interferon gamma activation of class II major histocompatibility complex genes. J Exp Med, 179:1605-1611.

Topletz AR, Le HN, Lee N, Chapman JD, Kelly EJ, Wang J, Isoherranen N. 2013. Hepatic Cyp2d and Cyp26a1 mRNAs and activities are increased during mouse pregnancy. Drug Metab Dispos, 41:312-319.

Wang F, Bei Y, Zhao Y, Song Y, Xiao J, Yang C. 2015. Telocytes in pregnancy-induced physiological liver growth. Cell Physiol Biochem, 36:250-258.

Williams Z. 2012. Inducing tolerance to pregnancy. N Engl J Med, 367:1159-1161

Wu X, Chen Y, Sun R, Wei H, Tian Z. 2012. Impairment of hepatic NK cell development in IFN-γ deficient mice. Cytokine, 60:616-625.

Xie H, Chen D, Luo X, Gao Z, Fang H, Huang J. 2013. Some characteristics of IL-5-producing T cells in mouse liver induced by Schistosoma japonicum infection. Parasitol Res, 112:1945-1951.

Yang L, Wang P, Mi H, Lv W, Liu B, Zhang L. 2019. Comparison of Th1 and Th2 cytokines production in ovine lymph nodes during early pregnancy. Theriogenology, 123:177-84.

Yang L, Wang Y, Li S, Zhu M, He K, Yao X, Zhang L. 2018. Differential expression of interferon-gamma, IL-4 and IL-10 in peripheral blood mononuclear cells during early pregnancy of the bovine. Reprod Biol, 18:312-315.

Yang L, Zhang LY, Qiao HY, Liu N, Wang YX, Li SJ. 2014. Maternal immune regulation by conceptus during early pregnancy in the bovine. Asian J Anim Vet Adv, 9:610-620.

Yoo I, Han J, Kim M, Jang H, Sa S, Choi SH, Ka H. 2017. Expression and regulation of interleukin 6 and its receptor at the maternal-conceptus interface during pregnancy in pigs. Theriogenology, 96:85-91.

Zhang L, Xia Y, Tang F, Li SJ, Yang L, Wang B. 2015. The regulation of intrauterine immune cytokines and chemokines during early pregnancy in the bovine. Large Anim Rev, 21:23-31

Zou Y, Hu M, Bao Q, Chan JY, Dai G. 2013. Nrf2 participates in regulating maternal hepatic adaptations to pregnancy. J Cell Sci, 126:1618-1625.

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