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生殖免疫学

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- 工作人员：杨颖
- 研究生：房文宁，宋志辉，孟朝阳，顾爱琴，杨丹丹，李丹丹

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　　生殖免疫学是由生殖生物学与免疫学两大学科 “ 融合 ” 的一门边缘学科。生殖免疫研究组是计划生育生殖生物学国家重点实验室所属的 18个研究组之一，主要从事生殖分子免疫学领域的研究工作。研究的兴趣集中在：(1)妊娠过程中免疫分子表达及调控机制。以生殖免疫调控基因和相关因子为切入点，探索母－胎免疫机制；(2)基因疫苗机理与应用的研究：以基因疫苗为技术平台，研究免疫控制生育、免疫抗生殖系统肿瘤的分子机理。本研究组近年来承担和已经完成的重要研究课题有：科技部九五攻关专题、 863 计划课题、中澳合作项目、中科院知识创新工程重要方向项目子课题、 科技部企业创新资助项目、滚动“ 863 ”计划课题、国家自然科学基金面上项目4项、973项目子课题和国家重大科学研究计划项目等，在母胎界面分子免疫调控机制、免疫控制生育和免疫抗生殖系统肿瘤的分子机理等方面取得了一些研究成果，近年来在国内外专业期刊发表了 SCI 论文 58 篇，申请和获得国家发明专利7项，建立了DNA 疫苗技术/免疫避孕/差异表达基因筛选与克隆等研究技术平台。



CX3CL1 facilitated peripheral NK cell migration. (a) Isolated splenic CD3？CD49b+ NK cells were spun onto slides, stained for CX3CR1 (green) or isotype IgG and counterstained with PI (red). Panels ii and vi show higher magnifications of the areas marked by the white rectangles in panels i and iii, respectively. Original magnification, × 63/1.40 (oil), zoom 1.00 (i and iii) and zoom 4.00 (ii and vi). Data shown are representative of two independent experiments from two mice. (b) To distinguish between chemotaxis and chemokinesis, NK cells were preincubated overnight with or without 500 ng/ml PTX before measuring cell migration. NK cells were gated on the basis of forward scatter and side scatter (FSC-SSC) and the numbers of NK cells were shown. Data show the mean±S.E.M. of three independent experiments. **P﹤0.01 by independent samples T-test. (c) After 12 h in culture, stromal cell CM (termed control CM), IFN-γ-treated stromal cell CM (termed IFN-γ CM), AG490-preincubated stromal cells before IFN-γ treatment CM (termed IFN-γ+AG490 CM) and AG490-treated stromal cell CM (termed AG490 CM) were collected and NK cell migration in response to them was measured as described above. For comparison, the chemotaxis index of NK cells toward control CM was set at 1. Data show the mean±S.E.M. of three independent experiments. *P﹤0.05 and **Po0.01 by one-way analysis of variance (ANOVA). (d) Five μg/ml anti-CX3CL1-neutralizing mAb or control rat IgG was added to the NK cell suspension and the migration of NK cells toward IFN-γ CM was measured as described above. The chemotaxis index of NK cells without any treatment toward IFN-γ CM was set at 1. Data show the mean ±S.E.M. of three independent experiments. **P﹤0.01 by one-way ANOVA



Further certainty, the regulation of cyp26a1 on Th17 cells at uterine implantation sites by uterine horn injection of anti-cyp26a1 antibody during mice peri-implantation. (A) Decrease in cyp26a1 levels at D5 uterine implantation sites following uterine horn injection of cyp26a1 antibody on D3 of pregnancy. Cyp26a1 levels were reduced at uterine implantation sites based on Western blotting and immunohistochemistry. b-actin was used as a loading control. Pairwise comparisons between each treatment group, *P < 0.05, **P < 0.01; bars = 50 lm. The data were derived from three separate samples from pregnant mice. (B) Th17cell increased in the peripheral blood and the spleen following uterine horn injection of cyp26a1 antibody. Flow cytometry histogram of Th17 cells is on the left side. Th17 cells are CD4+ RORct+. Th17 cell ratio was evaluated by flow cytometry analysis. The data are expressed as the% of CD4+ RORct+ double positive cells. The bars represent the SD of the mean. A paired t-test was used to assess the significance of differences. Pairwise comparisons between each treatment group, *P < 0.05. Three independent experiments were repeated for this time-point. A total of nine samples from pregnant mice were assessed. (C) Th17 levels were significantly reduced at uterine implantation sites based on Western blotting of pregnancy D5. b-actin was used as a loading control. The bars represent the SD of the mean of the relative value in grey (RORct/b-actin or IL-17/b-actin). A paired t-test was used to assess the significance of differences. Pairwise comparisons between each treatment group, *P < 0.05, **P < 0.01. The data were derived from three separate samples from pregnant mice. Three independent experiments were repeated for this time-point. A total of nine samples from pregnant mice were assessed. (D) Th17 cell levels were obviously reduced at implantation sites of D5 pregnancy based on immunohistochemistry of RORct and IL-17; bars = 200 and 25 lm. The data were derived from three separate samples from pregnant mice. At least, three independent experiments were repeated for this time-point. A total of nine samples from pregnant mice were assessed. E, embryo; S, uterine stroma; LE, Luminal epithelium; GE, Glandular epithelium.

 

 

研究内容和目标:
1. 母胎界面分子免疫调控机制
2. 免疫避孕研究
3. 生殖系统相关肿瘤侵润、转移等机理的研究

 

代表性发表论文:

  1. Quan-Hong Sun, Jing-Pian Peng*, Hong-Fei Xia, (2005)  Effect on expression of RT1-A and RT1-DM molecules treatment with interferon gamma at the maternal-fetal interface of pregnant rat. Hum. Reprod., 20(9):2639-2647.
  2. Shu-Qun Shi, Li Xu, Gang Zhao, Yang Ying, Jing-Pian Peng*,(2006) Apoptosis and tumor inhibition induced by human chorionic gonadotropin beta in mouse breast carcinoma. J Mol Med. 84:933–941.
  3. Hong-Fei Xia, Jing-Jing Ma, Jing Sun, Ying Yang, Jing-Pian Peng*(2010). Retinoic acid metabolizing enzyme CYP26A1 is implicated in rat embryo implantation Hum Reprod, 25(12):2985-2998.
  4. Bing-Chen Han, Hong-Fei Xia, Jing Sun, Ying Yang, Jing-Pian Peng*, (2010) Retinoic Acid-metabolizing Enzyme Cytochrome P450 26a1 (Cyp26a1) Is Essential for Implantation: Functional Study of Its Role in Early Pregnancy . J cell physiol.  223: 471-479.
  5. Zhong-Yin Li, Hu-He Chao, Hai-Yan Liu, Zhi-Hui Song, Li-Li Li, Yu-Jie Zhang, Ying Yang, Jing-Pian Peng,*(2014) IFN-γ induces aberrant CD49b+ NK cell recruitment through regulating CX3CL1: a novel mechanism by which IFN-γ provokes pregnancy failure, Cell Death Dis. 5, e1512; doi:10.1038/cddis.2014.470

 

 



IFN-γ treatment enhanced the accumulation of the CD49b+ NK cell subset. Syngeneically mated BALB/c female mice were injected with solvent or IFN-γ intraperitoneally on GD6 and killed on GD8. (a) Analysis of DBA lectin-stained uNK cells in the uteri by immunohistochemistry. Arrows indicate DBA lectin-positive cells. Photomicrographs are representative of five mice in each group. Panels iii and iv are higher magnifications of areas marked by the black rectangles in panels i and ii, respectively. Scale bar: 500 μm (i and ii) and 25 μm (iii and iv). (b) CD49b expression in uteri was analysed by quantitative PCR (top panel) and western blotting (bottom panel). Data show the mean± S.E.M. of four independent experiments and are obtained from four mice of each group, respectively. *Po0.05 by independent samples T-test. Flow cytometric analysis of cell suspensions from uteri (c) and peripheral blood (d). See Supplementary Figures 1B and C for the gating strategy and the percentages of CD3？CD49b+ NK cells (lowerright quadrant). Data show the mean±S.E.M. of four (uteri) or five (blood) independent experiments and are obtained from four (uteri) or five (blood) mice of each group, respectively. **Po0.01 by independent samples T-test. DB, decidua basalis; E, embryo