College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056021, China.
College of Animal Science and Technology, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China.
Anim Reprod Sci. 2020 Mar;214:106285. doi: 10.1016/j.anireprosci.2020.106285. Epub 2020 Jan 17.
Lymph nodes have functions in the adaptive immune response, and interferon-tau (IFNT), a primary pregnancy recognition signal in domestic ruminants has effects on immune regulation. It, however, is unclear whether early pregnancy induces an increase in the abundance of interferon-stimulated gene (ISG) mRNA transcripts and proteins in lymph nodes of sheep. In this study, lymph nodes were obtained on day 16 of the estrous cycle from non-pregnant ewes and days 13, 16 and 25 of gestation from pregnant ewes, and the abundance of ISG mRNA transcripts, including signal transducer and activator of transcription 1 (STAT1), phosphorylated STAT1 (p-STAT1), 2',5'-oligoadenylate synthetase (OAS1), myxovirus resistance protein 1 (MX1) and C-X-C motif chemokine 10 (CXCL10), was analyzed using real-time quantitative PCR. Furthermore, Western blot and immunohistochemistry analysis was conducted to assess relative abundance of proteins encoded by these genes. The results indicated that there was a larger abundance of STAT1 mRNA transcript and protein, and p-STAT1 protein in the maternal lymph node at days 16 and 25 of gestation, and that abundances of OAS1, MX1 and CXCL10 mRNA transcripts and protein were greatest on day 16 of gestations. In addition, STAT1 protein was located in the subcapsular sinus, lymph sinuses, B cells and T cells. The larger relative abundances of STAT1, p-STAT1, OAS1, MX1 and CXCL10 mRNA transcripts and/or protein in the lymph nodes of ewes may be associated with maternal immunoregulation through blood circulation and lymph circulation during early pregnancy.
淋巴结在适应性免疫反应中具有功能,而干扰素-τ(IFNT)是家畜妊娠识别的主要信号,对免疫调节有影响。然而,尚不清楚早期妊娠是否会导致绵羊淋巴结中干扰素刺激基因(ISG)mRNA 转录本和蛋白的丰度增加。在这项研究中,我们在发情周期的第 16 天从非妊娠母羊和妊娠的第 13、16 和 25 天获得淋巴结,并使用实时定量 PCR 分析包括信号转导和转录激活因子 1(STAT1)、磷酸化 STAT1(p-STAT1)、2',5'-寡腺苷酸合成酶 1(OAS1)、流感病毒抗性蛋白 1(MX1)和 C-X-C 基序趋化因子 10(CXCL10)在内的 ISG mRNA 转录本的丰度。此外,还进行了 Western blot 和免疫组织化学分析,以评估这些基因编码的蛋白质的相对丰度。结果表明,在妊娠第 16 和 25 天的母体淋巴结中,STAT1 mRNA 转录本和蛋白以及 p-STAT1 蛋白的丰度更大,而 OAS1、MX1 和 CXCL10 mRNA 转录本和蛋白的丰度在妊娠第 16 天最大。此外,STAT1 蛋白位于被膜下窦、淋巴窦、B 细胞和 T 细胞中。在妊娠早期,母羊淋巴结中 STAT1、p-STAT1、OAS1、MX1 和 CXCL10 mRNA 转录本和/或蛋白的相对丰度更大,这可能与母体内通过血液循环和淋巴循环进行的免疫调节有关。