Upregulation of Interferon-stimulated Genes and Interleukin-10 in Peripheral Blood Immune Cells During Early Pregnancy in Dairy Cows

Abstract

Abstract. In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP. Key words: Cow, Immune cells, Interferon tau, Pregnancy (J. Reprod. Dev. 58: [84][85][86][87][88][89][90] 2012) D uring the past 5 decades, milk production per cow has dramatically increased because of improved management, nutrition, and genetic selection [1,2]. In contrast, the decreases in fertility and conception rates of the modern high-producing dairy cow are the major causes of economic loss for dairy producers [2,3]. After establishment of pregnancy in domestic ruminants, the conceptus secretes interferon-tau (IFNT) as a maternal recognition factor [4]. IFNT acts in the uterus around day 16 after insemination and prevents luteolysis by inhibiting prostaglandin F 2α release, resulting in the maintenance of corpus luteum function; therefore, this period is termed the maternal recognition period (MRP) [5]. IFNT induces the synthesis and secretion of IFN-stimulated genes (ISGs) such as ISG15, 2′,5′-oligoadenylate synthetase (OAS-1), IFN regulatory factor 1, Mx1 and Mx2 not only in the uterus but also in blood cells in ewes and cows [6][7][8][9][10]. In fact, in bovine peripheral blood leukocytes, ISG15 mRNA levels were higher in pregnant cows than in nonpregnant cows on days 18 and 20 after artificial insemination (AI) [7][8][9]. Technology for the early detection of pregnancy is needed to identify nonpregnant cows and to synchronize and artificially inseminate these cows prior to the next ovulation, and many studies have focused on identify...

Interferon Stimulated Genes (Isgs): Novel Pregnancy Specific Biomarker In Buffaloes (Bubalus bubalis

“…In contrast, Shirasuna et al (2012) did not observe any change in expression pattern of ISG15 in pregnant and non-pregnant cows 25 . In a study conducted in cow, the ISG15 expression was shown to be significantly up-regulated on day 17 of the pregnancy 21 .…”

“…The OAS1 expression was amplified during days 15-18 in heifers 36 and cow 37 . Shirasuna et al (2012) documented that, the OAS1 expression did not vary in pregnant and nonpregnant cows 25 . The OAS1 mRNA expression was elevated within 24-48 h in PBMCs after initial signalling of IFNT in pregnant ewes 16 .…”

Interferon stimulated genes as peripheral diagnostic markers of early pregnancy in sheep: a critical assessment

“…Therefore real-time PCR analyses of ISG mRNA expression were run with different populations of circulating cells. Expression of ISG (ISG15, MX1, MX2) has been already described in PBMC (Shirasuna et al, 2012;Kizaki et al, 2013;Pugliesi et al, 2014), and in leucocytes (Yankey et al, 2001;Gifford et al, 2007;Green et al, 2010a). Kinetics of ISG expression level during peri-implantation was shown to be similar in PBMC and PMN (Shirasuna et al, 2012).…”

“…Expression of ISG (ISG15, MX1, MX2) has been already described in PBMC (Shirasuna et al, 2012;Kizaki et al, 2013;Pugliesi et al, 2014), and in leucocytes (Yankey et al, 2001;Gifford et al, 2007;Green et al, 2010a). Kinetics of ISG expression level during peri-implantation was shown to be similar in PBMC and PMN (Shirasuna et al, 2012). Kizaki et al (2013) demonstrated that ISG mRNA expression during early pregnancy is higher in granulocytes (eliminated during Percoll processing) than in monocytes and lymphocytes.…”

Maintenance or regression of the corpus luteum during multiple decisive periods of bovinepregnancy

“…In uterus, IFNT acts in a paracrine manner to prevent expression of estrogen receptor alpha and oxytocin receptor in endometrial luminal epithelium and superficial glandular epithelium, thereby altering release of luteolytic pulses of PGF (Spencer et al, 2007b). Interferon-tau also stimulates expression of specific genes, termed interferon-stimulated genes (Antoniazzi et al, 2013), in the uterus (Johnson et al, 1999) and in peripheral tissues such as the CL (Oliveira et al, 2008;Bott et al, 2010) and peripheral blood cells (Gifford et al, 2007;Shirasuna et al, 2012). Thus, circulating IFNT that escapes the uterine lumen might prevent CL regression by acting directly on CL and this action could be independent or synergistic with the actions of IFNT on uterine PGF secretion.…”