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Analysis of miRNA-mRNA expression profiles in maternal peripheral blood natural killer cells during pregnancy
Journal of Reproductive Immunology 112 (2015) 121–140
Contents lists available at ScienceDirect
Journal of Reproductive Immunology journal homepage: www.elsevier.com/locate/jreprimm
Organizing Secretariat of the 30th Annual Meeting of the Japan Society for Immunology of Reproduction – Oral Free Communication
01
02
Analysis of miRNA-mRNA expression profiles in maternal peripheral blood natural killer cells during pregnancy
Coexpression of TrkB and PD-L1 on human placenta
Yoichi Ishida 1,∗ , Dongwei Zhao 1 , Akihide Ohkuchi 1 , Tomoyuki Kuwata 1 , Shigeki Matsubara 1 , Shigeru Saito 3 , Toshihiro Takizawa 2 1
Department of Obstetrics and Gynecology, Jichi Medical University, Tochigi, Japan 2 Department of Molecular Medicine and Anatomy, Nippon Medical School, Tokyo, Japan 3 Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan Objective: We performed comprehensive miRNA and gene expression profiling of maternal peripheral blood natural killer (pNK) cells during pregnancy. Methods: Samples of pNK cells at different time points of pregnancy (first, second, and third trimester of gestation, and postdelivery) were obtained from healthy pregnant females with their informed consent. We performed real-time PCR-based array analysis to examine the expression levels of 756 miRNAs in maternal pNK cells using TaqMan Array MicroRNA Cards. Moreover, we performed DNA microarray analysis of gene expression levels in firstand third-trimester pNK cells using Agilent Microarray Kits. Results: PCR-based array analysis showed transfer of placentaderived miRNAs [chromosome 19 miRNA cluster (C19MC) miRNAs] into maternal pNK cells occurred during pregnancy. Rapid clearance of C19MC miRNAs also occurred in pNK cells after delivery. DNA microarray analysis identified NK cell function-related genes (e.g., KIR genes) that were differentially expressed between firstand third-trimester pNK cells. Conclusions: Maternal pNK cells may adjust to different gestational conditions via the uptake of exogenous miRNAs (e.g., C19MC miRNAs) during pregnancy. http://dx.doi.org/10.1016/j.jri.2015.09.006
Erina Nishigami 1,∗ , Rihito Kinami 1 , Hayato Terayama 2 , Kou Sakabe 2 , Hitoshi Ishimoto 3 , Mikio Mikami 3 , Yoshie Kametani 1 1 Department of Molecular Life Science, Tokai University School of Medicine, Kanagawa, Japan 2 Department of Anatomy and Histology, Division of Basic Medicine, Tokai University School of Medicine, Kanagawa, Japan 3 Department of Gynecology, Division of Specialized Medical Treatment, Tokai University School of Medicine, Kanagawa, Japan
Objective: Trophoblasts share characteristics of immunosuppression and invasion with solid tumors. However, whether invasion and immunosuppression is orchestrated is not clear. We examined the correlation between representative molecules, TrkB and PD-L1, together with the ligands and HLA-G. Materials and methods: Maternal blood (MB), fetal chorion and decidua were obtained from the full term placenta with the approval of the Tokai University Ethical Review Board. RNA was extracted and the molecular structure of TrkB was examined by RT-PCR. Localization of the cells expressing TrkB, BDNF, PD-L1 or PD-1 was analyzed by Immunohistchemistry (IHC). The expression was also analyzed by Flowcytemetry (FCM). Results and discussion: TrkB, BDNF and PD-L1 were highly expressed on syncytiotrophoblasts in the chorion. They were also expressed on the cells scattered in the decidua basialis and chorionic plate, although the types were not identified. TrkB molecules expressed on both chorion and decidua contained TrkB with intracellular tyrosine kinase (TK) domain, TrkB-T-Shc and TrkB-T1 without TK domain. PD-1 was expressed in MB and the blood vessels of the chorion. HLA-G was also expressed on 90% of macrophages in MB. The expression of TrkB and PD-L1 showed positive correlation in the HLA-G-positive macrophages. Both molecules on the placenta might be expressed in a coordinated manner. http://dx.doi.org/10.1016/j.jri.2015.09.007
0165-0378/$ – see front matter
Contents lists available at ScienceDirect
Journal of Reproductive Immunology journal homepage: www.elsevier.com/locate/jreprimm
Organizing Secretariat of the 30th Annual Meeting of the Japan Society for Immunology of Reproduction – Oral Free Communication
01
02
Analysis of miRNA-mRNA expression profiles in maternal peripheral blood natural killer cells during pregnancy
Coexpression of TrkB and PD-L1 on human placenta
Yoichi Ishida 1,∗ , Dongwei Zhao 1 , Akihide Ohkuchi 1 , Tomoyuki Kuwata 1 , Shigeki Matsubara 1 , Shigeru Saito 3 , Toshihiro Takizawa 2 1
Department of Obstetrics and Gynecology, Jichi Medical University, Tochigi, Japan 2 Department of Molecular Medicine and Anatomy, Nippon Medical School, Tokyo, Japan 3 Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan Objective: We performed comprehensive miRNA and gene expression profiling of maternal peripheral blood natural killer (pNK) cells during pregnancy. Methods: Samples of pNK cells at different time points of pregnancy (first, second, and third trimester of gestation, and postdelivery) were obtained from healthy pregnant females with their informed consent. We performed real-time PCR-based array analysis to examine the expression levels of 756 miRNAs in maternal pNK cells using TaqMan Array MicroRNA Cards. Moreover, we performed DNA microarray analysis of gene expression levels in firstand third-trimester pNK cells using Agilent Microarray Kits. Results: PCR-based array analysis showed transfer of placentaderived miRNAs [chromosome 19 miRNA cluster (C19MC) miRNAs] into maternal pNK cells occurred during pregnancy. Rapid clearance of C19MC miRNAs also occurred in pNK cells after delivery. DNA microarray analysis identified NK cell function-related genes (e.g., KIR genes) that were differentially expressed between firstand third-trimester pNK cells. Conclusions: Maternal pNK cells may adjust to different gestational conditions via the uptake of exogenous miRNAs (e.g., C19MC miRNAs) during pregnancy. http://dx.doi.org/10.1016/j.jri.2015.09.006
Erina Nishigami 1,∗ , Rihito Kinami 1 , Hayato Terayama 2 , Kou Sakabe 2 , Hitoshi Ishimoto 3 , Mikio Mikami 3 , Yoshie Kametani 1 1 Department of Molecular Life Science, Tokai University School of Medicine, Kanagawa, Japan 2 Department of Anatomy and Histology, Division of Basic Medicine, Tokai University School of Medicine, Kanagawa, Japan 3 Department of Gynecology, Division of Specialized Medical Treatment, Tokai University School of Medicine, Kanagawa, Japan
Objective: Trophoblasts share characteristics of immunosuppression and invasion with solid tumors. However, whether invasion and immunosuppression is orchestrated is not clear. We examined the correlation between representative molecules, TrkB and PD-L1, together with the ligands and HLA-G. Materials and methods: Maternal blood (MB), fetal chorion and decidua were obtained from the full term placenta with the approval of the Tokai University Ethical Review Board. RNA was extracted and the molecular structure of TrkB was examined by RT-PCR. Localization of the cells expressing TrkB, BDNF, PD-L1 or PD-1 was analyzed by Immunohistchemistry (IHC). The expression was also analyzed by Flowcytemetry (FCM). Results and discussion: TrkB, BDNF and PD-L1 were highly expressed on syncytiotrophoblasts in the chorion. They were also expressed on the cells scattered in the decidua basialis and chorionic plate, although the types were not identified. TrkB molecules expressed on both chorion and decidua contained TrkB with intracellular tyrosine kinase (TK) domain, TrkB-T-Shc and TrkB-T1 without TK domain. PD-1 was expressed in MB and the blood vessels of the chorion. HLA-G was also expressed on 90% of macrophages in MB. The expression of TrkB and PD-L1 showed positive correlation in the HLA-G-positive macrophages. Both molecules on the placenta might be expressed in a coordinated manner. http://dx.doi.org/10.1016/j.jri.2015.09.007
0165-0378/$ – see front matter