9 research outputs found
Endometrial factors similarly induced by IFNT2 and IFNTc1 through transcription factor FOXS1
<div><p>In ruminants, Interferon tau (IFNT) is the pregnancy recognition protein produced by the mononuclear trophectoderm of the conceptus, and is secreted into the uterine lumen during the peri-attachment period. In our previous study, the high-throughput RNA sequencing (RNA-seq) data obtained from bovine conceptuses during the peri-attachment period identified two <i>IFNT</i> mRNAs, <i>IFNT2</i> and <i>IFNTc1</i>. However, how each of these IFNT variants regulates endometrial gene expression has not been characterized. Using RNA-seq analysis, we evaluated how IFNT2 and IFNTc1 affected transcript expression in primary bovine endometrial epithelial cells (EECs). IFNT treatment induced 348 differentially expressed genes (DEGs); however, there are few DEGs in IFNT2 or IFNTc1 treated EECs, indicating that IFNT2-induced DEGs were similar to those induced by IFNTc1 treatment. In in silico analysis, we identified four IFNT2- and IFNTc1-induced pathways: 1) type II interferon signaling, 2) proteasome degradation, 3) type III interferon signaling, and 4) DNA damage response. We further demonstrated that IFNT2 and IFNTc1 up-regulated several transcription factors, among which forkhead box S1 (<i>FOXS1</i>) was identified as the most highly expressed gene. Furthermore, the knockdown of <i>FOXS1</i> in IFNT2- or IFNTc1-treated EECs similarly down-regulated 9 genes including <i>IRF3</i> and <i>IRF9</i>, and up-regulated 9 genes including <i>STAT1</i>, <i>STAT2</i>, and <i>IRF8</i>. These represent the first demonstration that effects of each IFNT on EECs were studied, and suggest that endometrial response as well as signaling mechanisms were similar between two IFNT variants existed in utero.</p></div
Transcriptional factors up-regulated by IFNTs in EECs.
<p>Transcriptional factors up-regulated by IFNTs in EECs.</p
Effects of FOXS1 knockdown on the expression of IFNTs downstream factors.
<p>(A-C) EECs were transfected with non-targeting control (Ctrl: 200 nM) or <i>FOXS1</i> siRNA (#1 or #2: 200 nM) for 48h, and then incubated with IFNT2 (B) or IFNTc1 (C) (2 x 10<sup>5</sup> cells/5000 IU/well) for 24 h. RNA was extracted from the EECs and subjected to real-time PCR analysis (n = 3 each group). <i>GAPDH</i> mRNA was used as an internal control for RNA integrity. Values represent the mean ± SEM from three independent experiments in each treatment.</p
Identification of gene expression induced by IFNTs in EECs.
<p>EECs were incubated without (Ctrl) or with IFNT2 or IFNTc1 (2 x 10<sup>5</sup> cells/5000 IU/well) for 24 h. RNA was extracted from the EECs and subjected to real-time PCR analysis to determine gene expression related to type II interferon, proteasome degradation, type III interferon, and DNA damage response signaling in Ctrl, IFNT2-, or IFNTc1-treated EECs (n = 3 each group). <i>GAPDH</i> mRNA was used as an internal control for RNA integrity. <sup>a</sup>P < 0.01, <sup>b</sup>P<0.05 vs. Ctrl. Value represent the mean ± SEM from three independent experiments in each treatment.</p
Differential gene expression in bovine endometrial epithelial cells treated with IFNT2 or IFNTc1.
<p>(A) Venn diagram shows the number of gene with 1.5-fold changes among Control (Ctrl), IFNT2, and IFNTc1 treatment groups, and right table lists increased or decreased genes in IFNT2 vs. IFNTc1 group, which overlap with Ctrl vs. IFNT2 or Ctrl vs. IFNTc1 group. (B) EECs were incubated without (Ctrl) or with IFNT2 or IFNTc1 (2 x 10<sup>5</sup> cells/5000 IU/well) for 24 h. RNA was extracted from the EECs and subjected to real-time PCR analysis on mRNA expression with overlapping IFNT2 vs. IFNTc1 group with other groups. <i>GAPDH</i> mRNA was used as an internal control for RNA integrity. Value represent the mean ± SEM from three independent experiments in each treatment. (C) these diagrams show pair plots comparison among Ctrl, IFNT2, and IFNTc1, and density plots in each groups. Figures show correlation coefficient among Ctrl, IFNT2, and IFNTc1.</p
Diagram illustrating the potential role of IFNT through FOXS1 in EECs.
<p>IFNT2 and IFNTc1 bind to their receptor and then activate STAT1 or STAT2. Activated STATs up-regulate FOXS1 expression, which down-regulates STATs expression, resulting in a negative feedback loop between STATs and FOXS1.</p
Genes related to IFNTs-induced enriched pathways in EECs.
<p>Genes related to IFNTs-induced enriched pathways in EECs.</p
Determination of IFNTs’ downstream transcription factors.
<p>EECs were incubated without (Ctrl) or with IFNT2 or IFNTc1 (2 x 10<sup>5</sup> cells/5000 IU/well) for 24 h. RNA was extracted from the EECs and subjected to real-time PCR analysis to determine the expression of transcription factors in Ctrl, IFNT2-, or IFNTc1-treated EECs (n = 3 each group). <i>GAPDH</i> mRNA was used as an internal control for RNA integrity. <sup>a</sup>P < 0.01, <sup>b</sup>P<0.05 vs. Ctrl. Value represent the mean ± SEM from three independent experiments in each treatment.</p
2012年から2016年にかけてナイジェリア南東部で検出されたラッサウイルスの遺伝学的解析
Lassa virus (LASV) is endemic in parts of West Africa where it causes Lassa fever (LF), a viral hemorrhagic fever with frequent fatal outcomes. The diverse LASV strains are grouped into six major lineages based on the geographical location of the isolated strains. In this study, we have focused on the lineage II strains from southern Nigeria. We determined the viral sequences from positive cases of LF reported at tertiary hospitals in Ebonyi and Enugu between 2012 and 2016. Reverse transcription-polymerase chain reaction (RT-PCR) showed that 29 out of 123 suspected cases were positive for the virus among which 11 viral gene sequences were determined. Phylogenetic analysis of the complete coding sequences of the four viral proteins revealed that lineage II strains are broadly divided into two genetic clades that diverged from a common ancestor 195 years ago. One clade, consisting of strains from Ebonyi and Enugu, was more conserved than the other from Irrua, although the four viral proteins were evolving at similar rates in both clades. These results suggested that the viruses of these clades have been distinctively evolving in geographically separate parts of southern Nigeria. Furthermore, the epidemiological data of the 2014 outbreak highlighted the role of human-to-human transmission in this outbreak, which was supported by phylogenetic analysis showing that 13 of the 16 sequences clustered together. These results provide new insights into the evolution of LASV in southern Nigeria and have important implications for vaccine development, diagnostic assay design, and LF outbreak management.長崎大学学位論文 学位記番号:博(医歯薬)甲第1107号 学位授与年月日:平成30年12月5日Author: Olamide K. Oloniniyi, Uche S. Unigwe, Sayaka Okada, Mayuko Kimura, Shota Koyano, Yukiko Miyazaki, Michael O. Iroezindu, Nnenna A. Ajayi, Chinedu M. Chukwubike, Nneka M. Chika-Igwenyi, Anne C. Ndu, Damian U. Nwidi, Haruka Abe, Shuzo Urata, Yohei Kurosaki, Jiro YasudaCitation: PLoS Neglected Tropical Diseases, 12(11), e0006971; 2018Nagasaki University (長崎大学)課程博