306 results on '"Joan S, Hunt"'
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2. Contemporary comparative placenta research--an interview with Allen Enders. Interview by Kent L Thornburg and Joan S Hunt.
- Author
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Enders A
- Subjects
- History, 20th Century, United States, Developmental Biology history
- Abstract
This Special Issue of The International Journal of Developmental Biology is dedicated to Allen C. Enders. His accomplishments are being honored because of his outstanding scientific contributions to our understanding of the development and mature structure of the mammalian placenta. He has consistently focused his research efforts on cutting edge questions related to the interaction of maternal and fetal tissues. While he has officially retired, he continues to study implantation and comparative placentology at the ultrastructural level. Enders has served as a link between the early placentologists who struggled to resolve the thinnest of layers in the placenta using light microscopy and modern day placentologists who now take advantage of confocal microscopy and electron microscopy to elucidate pathways for nutritive molecules as they pass from maternal to fetal compartments. Enders ground breaking electron micrographs will stand the test of time in revealing the relationships between embryonic tissue and maternal structures as they are variously modified in different species. The symbiotic interactions of genetically distinct tissues that form the placenta are required to nourish the fetus and perpetuate the species. This mysterious process makes mammalian reproduction the most exciting field of research in all of biology. The story of Allen Enders success will be an inspiration to all up and coming placentologists who are destined to discover the exciting world of placental biology.
- Published
- 2010
- Full Text
- View/download PDF
3. In Memoriam: Joan S. Hunt.
- Published
- 2019
- Full Text
- View/download PDF
4. Structural and Functional Basis for LILRB Immune Checkpoint Receptor Recognition of HLA-G Isoforms
- Author
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Haruki Matsubara, Hideo Fukuhara, Yuko Fukunaga, Yuji Sugita, Mitsunori Shiroishi, Katsumi Maenaka, R. Kanda, Atsushi Fukunaga, Joan S. Hunt, Toyoyuki Ose, Shunsuke Kita, Naoyuki Miyashita, Kaoru Hirose, Jun Kamishikiryo, and Kimiko Kuroki
- Subjects
Models, Molecular ,Gene isoform ,Protein Conformation ,Immunology ,Plasma protein binding ,Human leukocyte antigen ,Molecular Dynamics Simulation ,Ligands ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,LILRB2 ,HLA-G ,Humans ,Protein Isoforms ,Immunology and Allergy ,Receptors, Immunologic ,Binding site ,Receptor ,HLA-G Antigens ,Binding Sites ,Chemistry ,Immune checkpoint ,Cell biology ,Multiprotein Complexes ,beta 2-Microglobulin ,Protein Binding ,030215 immunology - Abstract
Human leukocyte Ig-like receptors (LILR) LILRB1 and LILRB2 are immune checkpoint receptors that regulate a wide range of physiological responses by binding to diverse ligands, including HLA-G. HLA-G is exclusively expressed in the placenta, some immunoregulatory cells, and tumors and has several unique isoforms. However, the recognition of HLA-G isoforms by LILRs is poorly understood. In this study, we characterized LILR binding to the β2-microglobulin (β2m)-free HLA-G1 isoform, which is synthesized by placental trophoblast cells and tends to dimerize and multimerize. The multimerized β2m-free HLA-G1 dimer lacked detectable affinity for LILRB1, but bound strongly to LILRB2. We also determined the crystal structure of the LILRB1 and HLA-G1 complex, which adopted the typical structure of a classical HLA class I complex. LILRB1 exhibits flexible binding modes with the α3 domain, but maintains tight contacts with β2m, thus accounting for β2m-dependent binding. Notably, both LILRB1 and B2 are oriented at suitable angles to permit efficient signaling upon complex formation with HLA-G1 dimers. These structural and functional features of ligand recognition by LILRs provide novel insights into their important roles in the biological regulations.
- Published
- 2019
5. Michael J. Soares Joan S. Hunt Placenta and Trophoblast: Methods and Protocols vol. 2 2007 Humana Press Totowa NJ, USA 452 pp.
- Author
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Bischof, P.
- Published
- 2007
- Full Text
- View/download PDF
6. Michael J. Soares Joan S. Hunt Placenta and Trophoblast Methods and Protocols vol. I 2006 Humana Press Totowa NJ, USA 1-58829-404-8, eBook ISBN 1-58829-983-4 470 pp., Hardcover, HC: USD 135.00
- Author
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Bischof, Paul
- Published
- 2007
- Full Text
- View/download PDF
7. Hormonal Regulation of Uterine Macrophages
- Author
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Joan S. Hunt, Lance Miller, and Jeralyn Sue Platt
- Subjects
Estrogen ,interferon-γ ,macrophage ,mouse ,nitric oxide ,progesterone. ,Immunologic diseases. Allergy ,RC581-607 - Published
- 1998
- Full Text
- View/download PDF
8. HLA Expression at the Maternal-Fetal Interface
- Author
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Heinz Hutter, Astrid Hammer, Gottfried Dohr, and Joan S. Hunt
- Subjects
MHC ,HLA class l ,placenta ,trophoblast ,immunology. ,Immunologic diseases. Allergy ,RC581-607 - Published
- 1998
- Full Text
- View/download PDF
9. Transcriptional regulation of VEGF-A by the unfolded protein response pathway.
- Author
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Rajarshi Ghosh, Kathryn L Lipson, Karen E Sargent, Arthur M Mercurio, Joan S Hunt, David Ron, and Fumihiko Urano
- Subjects
Medicine ,Science - Abstract
Angiogenesis is crucial to many physiological and pathological processes including development and cancer cell survival. Vascular endothelial growth factor-A (VEGFA) is the predominant mediator of angiogenesis in the VEGF family. During development, adverse environmental conditions like nutrient deprivation, hypoxia and increased protein secretion occur. IRE1alpha, PERK, and ATF6alpha, master regulators of the unfolded protein response (UPR), are activated under these conditions and are proposed to have a role in mediating angiogenesis.Here we show that IRE1alpha, PERK, and ATF6alpha powerfully regulate VEGFA mRNA expression under various stress conditions. In Ire1alpha(-/-) and Perk(-/-) mouse embryonic fibroblasts and ATF6alpha-knockdown HepG2 cells, induction of VEGFA mRNA by endoplasmic reticulum stress is attenuated as compared to control cells. Embryonic lethality of Ire1alpha-/- mice is due to the lack of VEGFA induction in labyrinthine trophoblast cells of the developing placenta. Rescue of IRE1alpha and PERK in Ire1alpha(-/-) and Perk(-/-) cells respectively, prevents VEGFA mRNA attenuation. We further report that the induction of VEGFA by IRE1alpha, PERK and ATF6 involves activation of transcription factors, spliced-XBP-1, ATF4 and cleaved ATF6 respectively.Our results reveal that the IRE1alpha-XBP-1, PERK-ATF4, and ATF6alpha pathways constitute novel upstream regulatory pathways of angiogenesis by modulating VEGF transcription. Activation of these pathways helps the rapidly growing cells to obtain sufficient nutrients and growth factors for their survival under the prevailing hostile environmental conditions. These results establish an important role of the UPR in angiogenesis.
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- 2010
- Full Text
- View/download PDF
10. IFPA Senior Award Lecture: Reproductive immunology in perspective – Reprogramming at the maternal–fetal interface
- Author
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Margaret G. Petroff and Joan S. Hunt
- Subjects
Fetus ,Pregnancy ,education.field_of_study ,Reproductive immunology ,Population ,Obstetrics and Gynecology ,Trophoblast ,Biology ,medicine.disease ,Article ,medicine.anatomical_structure ,Immune system ,Reproductive Medicine ,Placenta ,HLA-G ,embryonic structures ,Immunology ,medicine ,education ,Developmental Biology - Abstract
Involvement of the maternal and fetal immune systems in the events of pregnancy was generally overlooked by reproductive biologists until the mid-twentieth century when many landmark explorations were reported. Now, more than half a century later, it is well understood that with the initiation of pregnancy, immune cells in mammalian uteri are reprogrammed, losing their cytotoxic potential and providing an immunosuppressive environment suitable for harboring the genetically different fetus. We propose that it is the placenta that is mainly responsible for this conversion and maintenance throughout pregnancy. Studies in our laboratory indicate that trophoblast-derived soluble HLA-G has a subtle but well defined role in programming uterine placental macrophages, a potentially destructive immune cell population. Thus, placental HLA-G plays a critical role in assuring that the developing fetus emerges unscathed at parturition.
- Published
- 2013
11. Immunomodulatory molecules are released from the first trimester and term placenta via exosomes
- Author
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Susmita Jasti, S.M. Alam, Joan S. Hunt, Traci L. Nauser, Margaret G. Petroff, Melissa Gilliam, Herbert C. Hodes, C. Billstrand, and Sarika Kshirsagar
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B7 Antigens ,Placenta ,Pregnancy Trimester, Third ,Biology ,Exosomes ,B7-H1 Antigen ,Article ,Tissue Culture Techniques ,Immune system ,Cell-Derived Microparticles ,Pregnancy ,HLA-G ,medicine ,Humans ,Immunologic Factors ,Protein Isoforms ,Secretion ,Cells, Cultured ,HLA-G Antigens ,Cytotrophoblast ,Tetraspanin 30 ,Lysosome-Associated Membrane Glycoproteins ,Obstetrics and Gynecology ,Trophoblast ,Placentation ,Molecular biology ,Microvesicles ,Trophoblasts ,Cell biology ,Pregnancy Trimester, First ,medicine.anatomical_structure ,Reproductive Medicine ,Female ,Biomarkers ,Developmental Biology - Abstract
The semiallogenic fetus is tolerated by the maternal immune system through control of innate and adaptive immune responses. Trophoblast cells secrete nanometer scale membranous particles called exosomes, which have been implicated in modulation of the local and systemic maternal immune system. Here we investigate the possibility that exosomes secreted from the first trimester and term placenta carry HLA-G and B7 family immunomodulators. Confocal microscopy of placental sections revealed intracellular colocalization of B7-H1 with CD63, suggesting that B7-H1 associates with subcellular vesicles that give rise to exosomes. First trimester and term placental explants were then cultured for 24 hours. B7H-1 (CD274), B7-H3 (CD276) and HLA-G5 were abundant in pelleted supernatants of these cultures that contained microparticles and exosomes; the latter, however, was observed only in first trimester pellets and was nearly undetectable in term explant-derived pellets. Further purification of exosomes by sucrose density fractionation confirmed the association of these proteins specifically with exosomes. Finally, culture of purified trophoblast cells in the presence or absence of EGF suggested that despite the absence of HLA-G5 association with term explant-derived exosomes, it is present in exosomes secreted from mononuclear cytotrophoblast cells. Further, differentiation of cytotrophoblast cells reduced the presence of HLA-G5 in secreted exosomes. Together, the results suggest that the immunomodulatory proteins HLA-G5, B7-H1 and B7-H3, are secreted from early and term placenta, and have important implications in the mechanisms by which trophoblast immunomodulators modify the maternal immunological environment.
- Published
- 2012
12. Contemporary comparative placenta research - an interview with Allen Enders
- Author
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Joan S. Hunt and Kent L. Thornburg
- Subjects
Embryology ,medicine.anatomical_structure ,Evolutionary biology ,Electron micrographs ,Placenta ,Fetal tissue ,medicine ,Zoology ,Biology ,Developmental Biology - Abstract
This Special Issue of The International Journal of Developmental Biology is dedicated to Allen C. Enders. His accomplishments are being honored because of his outstanding scientific contributions to our understanding of the development and mature structure of the mammalian placenta. He has consistently focused his research efforts on cutting edge questions related to the interaction of maternal and fetal tissues. While he has officially retired, he continues to study implantation and comparative placentology at the ultrastructural level. Enders has served as a link between the early placentologists who struggled to resolve the thinnest of layers in the placenta using light microscopy and modern day placentologists who now take advantage of confocal microscopy and electron microscopy to elucidate pathways for nutritive molecules as they pass from maternal to fetal compartments. Enders’ ground breaking electron micrographs will stand the test of time in revealing the relationships between embryonic tissue and maternal structures as they are variously modified in different species. The symbiotic interactions of genetically distinct tissues that form the placenta are required to nourish the fetus and perpetuate the species. This mysterious process makes mammalian reproduction the most exciting field of research in all of biology. The story of Allen Enders’ success will be an inspiration to all up and coming placentologists who are destined to discover the exciting world of placental biology.
- Published
- 2010
13. The Role of Integrins in Reproduction
- Author
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Jeffery A. Bowen and Joan S. Hunt
- Subjects
General Biochemistry, Genetics and Molecular Biology - Published
- 2008
14. A Commentary on Gestational Programming and Functions of HLA-G in Pregnancy
- Author
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Asgerally T. Fazleabas, Judith L. Pace, Daudi K. Langat, Pedro J. Morales, and Joan S. Hunt
- Subjects
Gene isoform ,medicine.drug_class ,Placenta ,Context (language use) ,Human leukocyte antigen ,Biology ,Monoclonal antibody ,Article ,Antigen ,HLA Antigens ,Pregnancy ,HLA-G ,medicine ,Humans ,Protein Isoforms ,RNA, Messenger ,Receptors, Immunologic ,HLA-G Antigens ,Genetics ,Histocompatibility Antigens Class I ,Obstetrics and Gynecology ,Trophoblast ,Trophoblasts ,Blastocyst ,Receptors, KIR2DL5 ,medicine.anatomical_structure ,Reproductive Medicine ,Female ,Developmental Biology - Abstract
HLA-G is an HLA class Ib gene that is highly expressed in human trophoblast cells. The single HLA-G mRNA is alternatively spliced to generate at least seven transcripts, three of which encode soluble isoforms. Many studies have shown that high levels of soluble antigens are associated with successful implantation and graft acceptance. To study expression, regulation and functions of two of the soluble isoforms, HLA-G5 and HLA-G6, we generated recombinant proteins in eukaryotic cells and developed monoclonal antibodies specific for each of the two proteins. In addition, we investigated the olive baboon Paan-AG gene as a potential functional correlate of HLA-G. Here, we present summaries of the studies that have been conducted in our laboratory using these tools and discuss the results within the context of the research on this topic that is ongoing in ours and other laboratories worldwide. Collectively, the data indicate that soluble HLA-G is a critical contributor to immune privilege in pregnancy and imply that this placenta-derived substance may impact other pathways leading to successful reproduction.
- Published
- 2007
15. Contributors
- Author
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Kjersti M. Aagaard, David F. Albertini, Eugene D. Albrecht, Steven M. Anderson, William E. Armstrong, Richard J. Auchus, Susan P. Bagby, Jacques Balthazart, April K. Binder, Jeffrey D. Blaustein, S. Marc Breedlove, Paula J. Brunton, Jeeyeon Cha, Shawn L. Chavez, Paula E. Cohen, Gerard S. Conway, John F. Couse, Geert J. de Vries, Emily DeFranco, Francesco J. DeMayo, Sudhansu K. Dey, Juan M. Dominguez, Edward M. Eddy, Rafael A. Fissore, Loretta M. Flanagan-Cato, Harvey M. Florman, Nancy G. Forger, Douglas L. Foster, Stephen Franks, Jennifer R. Gardiner, Kathrin Gassei, George D. Giraud, Robert L. Goodman, Andrea C. Gore, David R. Grattan, Janet E. Hall, Kate Hardy, Frances J. Hayes, David Hazlerigg, Mark P. Hedger, Jon D. Hennebold, Allan E. Herbison, Sylvia C. Hewitt, Stanley M. Hileman, Barry T. Hinton, J. Kim Holloway, Elaine M. Hull, Joan S. Hunt, Mary Hunzicker-Dunn, E. Keith Inskeep, Thomas Jansson, Sherri L. Jones, Kenneth S. Korach, Michael J. Large, Jon E. Levine, Xiao-Feng Li, Hyunjung (Jade) Lim, Mark Lindgren, Zhilin Liu, Joseph S. Lonstein, Paul S. MacLean, Catherine A. Marler, Kelly Mayo, Erik C. Mazur, Craig A. McArdle, Margaret M. McCarthy, Neil J. McKenna, James L. McManaman, Sam Mesiano, Joan I. Morrell, Louis J. Muglia, Makoto C. Nagano, Margaret C. Neville, Kevin T. O’Byrne, Peter O’Shaughnessy, Kyle E. Orwig, Stephanie A. Pangas, Gerald J. Pepe, Mariana Pereira, Margaret G. Petroff, James G. Pfaus, Bart T. Phillips, Tony M. Plant, Vincent Prevot, Gail S. Prins, Aleksandar Rajkovic, Cyril Ramathal, Renee A. Reijo Pera, JoAnne S. Richards, Emilie F. Rissman, Bernard Robaire, Mark S. Roberson, Sarah A. Robertson, John A. Russell, Yoel Sadovsky, Masayuki Shimada, Valerie Simonneaux, Lee B. Smith, Richard L. Stouffer, Susan S. Suarez, Melissa A. Suter, Amanda Swain, Manuel Tena-Sempere, Ei Terasawa, Kent L. Thornburg, Paul Le Tissier, Kiyotaka Toshimori, Hanna Valli, William H. Walker, Wipawee Winuthayanon, Selma Feldman Witchel, Larry J. Young, and Anthony J. Zeleznik
- Published
- 2015
16. Immunology of Pregnancy
- Author
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Margaret G. Petroff, Joan S. Hunt, and Sarah A. Robertson
- Subjects
biology ,Regulatory T cell ,Human leukocyte antigen ,Dendritic cell ,biochemical phenomena, metabolism, and nutrition ,Major histocompatibility complex ,Acquired immune system ,Natural killer cell ,Immune tolerance ,medicine.anatomical_structure ,Immune system ,embryonic structures ,Immunology ,medicine ,biology.protein ,bacteria - Abstract
The principal role of the immune system is to defend the integrity of the host by preventing infectious disease, tumor development, and contamination of the host genome. Mammalian pregnancy, which features gametes and embryos expressing proteins derived from disparate maternal and paternal genes, presents a serious challenge to the maternal immune system. A combination of strategies is involved in negotiating and preventing fetal immune rejection. Specific aspects of the immune response are circumvented, while others are engaged and persuaded to permit the intimate associations between maternal and fetal cells that characterize embryo development, implantation, and placental development. In this chapter, features of the female reproductive organs and specific strategies that protect against immune-mediated embryonic loss are described. Key concepts are that both the mother and the fetus contribute to successful pregnancy, that multiple immune evasion and tolerogenic mechanisms are deployed, and that immune adaptations to permit internalization of the fetus depend on ovarian steroid hormones and are initiated from the time of conception. Major features include (1) specific structural elements of the reproductive tissues that support immune accommodation, (2) characteristic features of resident leukocyte subpopulations, (3) immune suppressive and pro-tolerogenic molecules produced by maternal and fetal cells, (4) restricted fetal trophoblast expression of major histocompatibility molecules, and (5) a critical role for T-regulatory cells which suppress inflammation and inhibit maternal immunity to the fetus.
- Published
- 2015
17. Differential expression of human leukocyte antigen-G (HLA-G) messenger RNAs and proteins in normal human prostate and prostatic adenocarcinoma
- Author
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Asgerally T. Fazleabas, J. Sue Platt, Ossama Tawfik, Joan S. Hunt, and Daudi K. Langat
- Subjects
Male ,PCA3 ,Pathology ,medicine.medical_specialty ,Immunology ,Human leukocyte antigen ,Adenocarcinoma ,Biology ,HLA Antigens ,Prostate ,HLA-G ,Leukocytes ,medicine ,Humans ,Immunology and Allergy ,RNA, Messenger ,Cellular localization ,HLA-G Antigens ,Histocompatibility Antigens Class I ,Prostatic Neoplasms ,Obstetrics and Gynecology ,Hyperplasia ,medicine.disease ,Immunohistochemistry ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Reproductive Medicine ,Health - Abstract
Human leukocyte antigen-G (HLA-G) is a major histocompatibility complex class Ib gene expressed in normal organs and in some tumors. The glycoproteins encoded by this gene are best known for their immunosuppressive properties. Because isoform-specific expression of HLA-G in male reproductive organs has not been reported, we investigated HLA-G1, -G2, -G5, -G6 mRNAs and proteins in four-to-five samples of normal prostate glands, prostates with benign prostatic hyperplasia and prostate adenocarcinomas using RT-PCR and immunohistochemistry. All tissues contained HLA-G1, -G2, -G5 and -G6 specific mRNAs, but only HLA-G5 protein was detectable. In normal prostate glands, HLA-G5 protein was prominent in the cytoplasm of tubuloglandular epithelia and in glandular secretions. Staining was reduced in samples of benign prostatic hyperplasia but remained localized to the cytoplasm of glandular epithelia and secretions. In prostatic adenocarcinomas, HLA-G5 protein was detectable mainly in the secretions. Thus, HLA-G5 but not HLA-G1, -G2 or -G6 is produced in the normal prostate and is present in prostatic secretions. In addition, normal cellular localization is disturbed in benign and malignant prostatic adenocarcinomas. The results are consistent with this molecule may influencing female immune receptivity to sperm and suggest that such immunosuppression could be disturbed in men with prostatic adenocarcinomas.
- Published
- 2006
18. FLICE-inhibitory Protein: Expression in Early and Late Gestation Human Placentas
- Author
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Joan S. Hunt and H. Ka
- Subjects
Adult ,Gene isoform ,Programmed cell death ,Term Birth ,Cellular differentiation ,CASP8 and FADD-Like Apoptosis Regulating Protein ,Extraembryonic Membranes ,Gestational Age ,Biology ,Immunoenzyme Techniques ,Pregnancy ,Cell Line, Tumor ,Placenta ,Gene expression ,medicine ,Humans ,Protein Isoforms ,Choriocarcinoma ,RNA, Messenger ,Caspase 8 ,Cytotrophoblast ,Reverse Transcriptase Polymerase Chain Reaction ,Intracellular Signaling Peptides and Proteins ,Gene Expression Regulation, Developmental ,Obstetrics and Gynecology ,Trophoblast ,Trophoblasts ,Cell biology ,Pregnancy Trimester, First ,medicine.anatomical_structure ,Reproductive Medicine ,Flip ,Caspases ,embryonic structures ,Immunology ,Female ,Developmental Biology - Abstract
The apoptosis cascade that plays a central role in normal and pathological processes is strictly controlled, in part by FLIP (Fas-associated death domain-like interleukin-1beta-converting enzyme-inhibitory protein), an inhibitor of caspase-8. Here, we report the expression of long and short isoforms of FLIP mRNAs and proteins in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the FLIP gene in all samples. Analysis by immunoblotting revealed that both long and short forms of FLIP proteins are present in early and late gestation human placentas with increasing levels over gestation and that FLIP proteins are present in normal and transformed trophoblast cells. Immunohistochemical experiments performed on paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy demonstrated that FLIP proteins are present in caspase-8-expressing cells and that expression patterns of FLIP differed according to cell lineage and stage of cell differentiation. The results of this study are consistent with the postulate that FLIP proteins have critical roles in placental cell survival and suggest that FLIP may protect normal and transformed trophoblast cells from cell death.
- Published
- 2006
19. Trophoblast CD274 (B7-H1) Is Differentially Expressed Across Gestation: Influence of Oxygen Concentration1
- Author
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Lesya M. Holets, Margaret G. Petroff, and Joan S. Hunt
- Subjects
Fetus ,Trophoblast ,Cell Biology ,General Medicine ,Biology ,medicine.disease ,Preeclampsia ,Oxygen tension ,Andrology ,medicine.anatomical_structure ,Syncytiotrophoblast ,Reproductive Medicine ,Downregulation and upregulation ,Placenta ,embryonic structures ,Immunology ,medicine ,Limiting oxygen concentration - Abstract
Modulation of the maternal immune system by the placenta is a mechanism by which the fetus ensures its own survival in a genetically foreign environment. The immunoinhibitor CD274 (also called B7-H1 or PD-L1) is highly expressed in the placenta, positioned to interact with maternal leukocytes. Further, immunoblot analysis of first- and second-trimester placental lysates showed that CD274 expression is low in the first trimester but dramatically rises around the onset of the second trimester. As this coincides with the expected onset of maternal blood flow to the placenta and a corresponding rise in local oxygen tension, we explored the possibility that oxygen regulates CD274 expression in trophoblast cells by culturing term trophoblast cells under oxygen concentrations similar to those found in vivo. Indeed, CD274 protein levels paralleled the in vivo situation: expression increased with rising oxygen concentrations. Furthermore, downregulation of CD274 mRNA by low oxygen was rapid, occurring within 4–12 h. We conclude that oxygen is a potential mediator of CD274 expression in vivo such that it is induced coincidentally on exposure of fetal tissues to maternal blood. Further, the regulation of this immunomodulator by oxygen may implicate its alteration during and involvement in the pathogenesis of complications of pregnancy such as preeclampsia.
- Published
- 2006
20. Antigen presenting cells and HLA-G – a review
- Author
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Joan S. Hunt and R.H. McIntire
- Subjects
Stromal cell ,Antigen presentation ,Antigen-Presenting Cells ,Apoptosis ,HLA Antigens ,Pregnancy ,Decidua ,Immune Tolerance ,Humans ,Receptors, Immunologic ,Antigen-presenting cell ,Maternal-Fetal Exchange ,HLA-G Antigens ,Innate immune system ,CD40 ,Follicular dendritic cells ,biology ,Macrophages ,Histocompatibility Antigens Class I ,Models, Immunological ,Obstetrics and Gynecology ,Dendritic Cells ,Dendritic cell ,B-1 cell ,Phenotype ,Reproductive Medicine ,Immunology ,biology.protein ,Cytokines ,Female ,Developmental Biology - Abstract
Maternal antigen presenting cells, which are macrophages and dendritic cells, are scattered throughout human decidualized endometrium during all stages of pregnancy. These powerful, multi-functional leukocytes reside in close proximity to uterine glandular epithelium, uterine blood vessels, and HLA-G-producing invasive cytotrophoblast cells. Macrophages and dendritic cells, which express the HLA-G receptors, ILT2 and ILT4, play major roles in driving innate and adaptive immune responses, altering the behavior of local stromal cells, shaping the cytokine microenvironment, and protecting the tissue from infection. Therefore, encounters between decidual antigen presenting cells and HLA-G molecules are likely to influence uterine and placental homeostasis as well as local maternal immune responses to the fetus during pregnancy.
- Published
- 2005
21. B7 Family Molecules Are Favorably Positioned at the Human Maternal-Fetal Interface1
- Author
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Joan S. Hunt, Margaret G. Petroff, Peter Sedlmayr, Teresa A. Phillips, Lieping Chen, and Dagmar Azzola
- Subjects
medicine.medical_specialty ,Cytotrophoblast ,Stromal cell ,Antigen presentation ,Trophoblast ,Cell Biology ,General Medicine ,Biology ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,Syncytiotrophoblast ,Reproductive Medicine ,Placenta ,Internal medicine ,embryonic structures ,medicine ,Cytotrophoblasts ,Northern blot - Abstract
The human placenta utilizes both active and passive mechanisms to evade rejection by the maternal immune system. We investigated the pattern of expression of the B7 family of immunomodulatory molecules B7-H1 (PD-L1), B7-2 (CD86), and B7-1 (CD80) at the term maternal-fetal interface. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that B7-H1 mRNA is abundant in term placenta and that cytotrophoblasts are sources of this message. Immunohistochemistry demonstrated that B7-H1 is constitutively expressed by the syncytiotrophoblast and by extravillous cytotrophoblasts, both of which are juxtaposed to maternal blood and tissue. By contrast, placental stromal cells, including macrophages, lacked the protein. Expression of B7-H1 protein was low in first-trimester placenta compared to second- and third-trimester tissue (P < 0.05) and was enhanced in cultured cytotrophoblasts by treatment with either interferon-gamma or epidermal growth factor (P < 0.05), suggesting that one or both of these mediators regulates B7-H1 expression in the placenta. RT-PCR and immunofluorescence analysis of term placental tissue revealed different patterns of expression of the immunostimulatory protein, B7-2. In contrast to B7-H1, B7-2 mRNA and protein were absent in cytotrophoblast cells but present in maternal macrophages and some fetal macrophages. The B7-1 mRNA and protein were absent at the maternal-fetal interface. These studies document expression of the B7 family proteins at the maternal-fetal interface and demonstrate that B7-H1 is positioned such that it could facilitate protection of fetal cells against activated maternal leukocytes. Conversely, B7-2 was absent on trophoblasts and was appropriately localized to fetal and maternal macrophages, which may participate in antigen presentation.
- Published
- 2003
22. Do Nonhuman Primates Comprise Appropriate Experimental Models for Studying the Function of Human Leukocyte Antigen-G?1
- Author
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Daudi K. Langat and Joan S. Hunt
- Subjects
Genetics ,biology ,Trophoblast ,Cell Biology ,General Medicine ,Human leukocyte antigen ,Major histocompatibility complex ,In vitro ,Cell biology ,medicine.anatomical_structure ,Reproductive Medicine ,MHC class I ,Gene expression ,medicine ,biology.protein ,Gene ,CD8 - Abstract
The expression and function of the human major histocompatibility complex (MHC) class Ia genes, human leukocyte antigen (HLA)-A, -B, and -C, is well-established; they are expressed in most nucleated cells and present endogenous peptides to CD8+ T cells. However, MHC class Ib genes are poorly characterized and have unknown functions. In humans, the best-characterized class Ib gene is HLA-G. This gene has a restricted tissue expression of the mRNA and a unique pattern of protein expression; it is expressed mainly in the extravillous cytotrophoblast cells in the placenta. The function of HLA-G is not clear, but its presence at the maternal-fetal interface suggests a role in protection of the semiallogeneic fetus. Whereas functional studies using in vitro models and transgenic mice provide useful insights regarding the potential function of this molecule, in vivo studies cannot be performed in humans. Nonhuman primates that are closely related to humans phylogenetically contain homologues of HLA-G. T...
- Published
- 2002
23. Baboon placentas express soluble and membrane-bound Paan-AG proteins encoded by alternatively spliced transcripts of the class Ib major histocompatibility complex gene, Paan-AG
- Author
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Asgerally T. Fazleabas, Daudi K. Langat, Jason M. Mwenda, Pedro J. Morales, and Joan S. Hunt
- Subjects
Transcription, Genetic ,Placenta ,Molecular Sequence Data ,Immunology ,Genes, MHC Class I ,Human leukocyte antigen ,Major histocompatibility complex ,Exon ,HLA Antigens ,Genetics ,Animals ,Protein Isoforms ,Tissue Distribution ,Amino Acid Sequence ,RNA, Messenger ,Gene ,Sequence Deletion ,HLA-G Antigens ,Base Sequence ,biology ,Histocompatibility Antigens Class I ,Alternative splicing ,Intron ,Molecular biology ,Stop codon ,Alternative Splicing ,RNA splicing ,biology.protein ,Sequence Alignment ,Papio - Abstract
The human class Ib major histocompatibility complex (MHC) molecule, HLA-G, is unique in its limited polymorphism, high expression in the placenta and generation of multiple transcripts by alternative splicing. The proteins encoded by these transcripts are believed to modulate maternal-fetal immunological relationships during pregnancy. The baboon placenta contains messages encoded by a novel MHC gene, Paan-AG, which is evolutionarily related to the HLA-A locus, but shares unique characteristics with HLA-G. In this study, we show that the Paan-AG message is alternatively spliced to generate at least seven transcripts. One of these transcripts retains intron 4 and encodes a soluble glycoprotein with three external domains and a unique 21-amino-acid sequence at the carboxyl terminus, similar to soluble HLA-G1. This glycoprotein was detected in first trimester placental villous cytotrophoblast and syncytiotrophoblast, and in extravillous cytotrophoblast cells in the basal plate in term placenta. Four of the transcripts ( Paan-AG1, Paan-AG2, Paan-AG3, Paan-AG4) encode membrane-bound class Ib MHC glycoprotein isoforms. Paan-AG1 protein expression was similar to that of sPaan-AG, while Paan-AG2 protein was not detected in these tissues. The other two transcripts ( Paan-AGx and Paan-AGxi) contain a truncated exon 3 and multiple stop codons. Paan-AG1 and Paan-AGx transcripts were detected in a number of non-placental tissues, but these transcripts contained multiple stop codons. Because of the structural similarities and common features of organ-specific expression and splicing of the message, studies on Paan-AG may be of value in dissecting the functions of the class Ib proteins in human pregnancy.
- Published
- 2002
24. B7 Family Molecules: Novel Immunomodulators at the Maternal-Fetal Interface
- Author
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Teresa A. Phillips, Joan S. Hunt, Margaret G. Petroff, and Lieping Chen
- Subjects
Adult ,Gestational Age ,Biology ,Response Elements ,B7-H1 Antigen ,law.invention ,Interferon-gamma ,Immune system ,Syncytiotrophoblast ,Adjuvants, Immunologic ,Antigens, CD ,Pregnancy ,law ,Interferon ,Placenta ,Tumor Cells, Cultured ,medicine ,Humans ,Maternal fetal ,Choriocarcinoma ,RNA, Messenger ,Maternal-Fetal Exchange ,reproductive and urinary physiology ,Messenger RNA ,Membrane Glycoproteins ,Dose-Response Relationship, Drug ,Obstetrics and Gynecology ,Trophoblast ,Blood Proteins ,Recombinant Proteins ,Trophoblasts ,Disease Models, Animal ,medicine.anatomical_structure ,Reproductive Medicine ,embryonic structures ,Immunology ,B7-1 Antigen ,Recombinant DNA ,Female ,Peptides ,Developmental Biology ,medicine.drug - Abstract
The placenta utilizes both active and passive mechanisms to evade rejection by the maternal immune system. Recently, the mRNA for two newly cloned members of the B7 family of immunomodulatory cell-associated proteins have been identified in the human term placenta. In this article, we review the current knowledge of the B7 family member B7-H1, and discuss how it may participate in modulation of the maternal immune system at the maternal-fetal interface. B7-H1 has been found to possess immunostimulatory or immunoinhibitory properties, and immunohistological examination of first trimester and term placenta has revealed that this protein is abundant in the placenta. B7-H1 is highly expressed by both the syncytiotrophoblast and extravillous cytotrophoblast, both of which lie in direct contact with maternal blood and tissue. Further, treatment of the choriocarcinoma cell line, JEG-3, with recombinant human interferon (IFN)-γ resulted in a dose-dependent increase in the abundance of the message for B7-H1, suggesting that IFN-γ could regulate expression of B7-H1 by the trophoblast. These studies document that the positioning of B7-H1 at the maternal-fetal interface is such that it could participate in suppression of activated maternal leukocytes.
- Published
- 2002
25. Placenta
- Author
-
Joan S. Hunt and Kent L. Thornburg
- Subjects
Embryology ,Developmental Biology - Published
- 2010
26. Eosinophils are cellular targets of the novel uteroplacental heparin-binding cytokine decidual/trophoblast prolactin-related protein
- Author
-
Ryuta Ishimura, Guoli Dai, N A Lee, Joan S. Hunt, D S Walia, Michael J. Soares, J J Lee, Heiner Müller, and D Wang
- Subjects
medicine.medical_specialty ,Placenta ,Recombinant Fusion Proteins ,Endocrinology, Diabetes and Metabolism ,Cell Culture Techniques ,Mice, Inbred Strains ,CHO Cells ,Pregnancy Proteins ,Biology ,Mice ,Endocrinology ,Pregnancy ,Fetal membrane ,Cricetinae ,Internal medicine ,medicine ,Animals ,Decidual cells ,Heparin ,Uterus ,Decidua ,Trophoblast ,Estrogens ,Eosinophil ,Prolactin ,Rats ,Eosinophils ,Chorioallantoic membrane ,medicine.anatomical_structure ,Cell culture ,Female - Abstract
The uterus and placenta of the mouse and rat produce a member of the prolactin (PRL) family referred to as decidual/trophoblast PRL-related protein (d/tPRP). This cytokine/hormone has been hypothesized to regulate decidual cell activities needed for the establishment and maintenance of gestation. An alkaline phosphatase (AP)-tagging strategy was used to identify d/tPRP target cells. AP-d/tPRP bound to virtually all cells and tissues to which it was exposed, consistent with our earlier evidence that d/tPRP binds to heparin-containing molecules. Moreover, we found that co-incubation with heparin or pretreatment with heparitinase greatly decreased the binding of AP-d/tPRP to tissue sections. In addition, we observed that the AP-d/tPRP probe bound to the surface of Chinese hamster ovary (CHO) cells but not to heparan sulfate-deficient CHO-pgsD-677 cells. Potential unique non-heparin d/tPRP binding sites within mouse and rat uteroplacental tissues were identified by consecutively incubating sections with AP-d/tPRP followed by heparin. This strategy led to the identification of d/tPRP target cells associated with the uterus and the labyrinth zone of the chorioallantoic placenta. Within the uterus, d/tPRP specifically bound to eosinophils. d/tPRP-binding and eosinophil peroxidase activity were co-localized and showed similar patterns of distribution during the estrous cycle, pregnancy, and following hormonal manipulation. d/tPRP interactions with eosinophils were further demonstrated in the lung and intestine, with eosinophils isolated from the peritoneum, and in mice with generalized tissue eosinophilia. Collectively, these findings suggest that intercellular d/tPRP targeting is mediated through associations with heparin-containing molecules which help direct d/tPRP to specific interactions with eosinophils within the uterus and with the labyrinthine compartment of the chorioallantoic placenta.
- Published
- 2000
27. Nitric Oxide Synthase-2 and Expression of Perforin in Uterine NK Cells
- Author
-
Joan S. Hunt and Tim G. Burnett
- Subjects
Pore Forming Cytotoxic Proteins ,Pathology ,medicine.medical_specialty ,Immunology ,Uterus ,Down-Regulation ,Nitric Oxide Synthase Type II ,Mice ,Pregnancy ,medicine ,Animals ,Immunology and Allergy ,Embryo Implantation ,Lymphocyte Count ,RNA, Messenger ,Northern blot ,Mice, Knockout ,Messenger RNA ,Membrane Glycoproteins ,biology ,Perforin ,Nitric oxide synthase 2 ,Molecular biology ,Killer Cells, Natural ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Granzyme ,embryonic structures ,biology.protein ,Immunohistochemistry ,Female ,Metrial Gland ,Nitric Oxide Synthase - Abstract
In human, mouse, and rat pregnancy, maternal NK cells accumulate and differentiate at implantation sites. These cells, termed uterine NK (uNK) cells, express NO synthase (NOS)-2 and develop cytolytic molecules such as perforin and granzymes during differentiation in situ. In this study, relationships between expression of the NOS-2 gene, uNK cell population density and tissue distribution, and synthesis of perforin were investigated. Uteri from wild-type (WT) and NOS-2−/− mice were collected at gestation days (g.d.) 8, 10, 12, 14, and 16 (n, >2/g.d.). Histochemical staining failed to reveal any differences between the population densities or tissue distributions of uNK cells in WT and NOS-2−/− uteri at any stage of gestation. By contrast, immunohistochemical staining with anti-perforin Abs demonstrated significantly fewer perforin-positive uNK cells in two uterine compartments of NOS-2−/− mice in comparison to the same compartments in WT mouse uteri. Perforin-positive uNK cells were reduced in NOS-2−/− metrial glands at g.d. 8, 10, and 12 and in decidua basalis at g.d. 12 (p < 0.05). Analysis of perforin protein by immunoblotting confirmed this observation. Northern blot hybridization studies showed that loss of perforin protein in NOS-2−/− mice was accompanied by decreased steady-state levels of perforin mRNA. These results demonstrate that migration of uNK cells into the uterus, selection of residency sites, and proliferation in situ are independent of NOS-2. By contrast, their differentiation, including transcription and translation of the cytotoxic molecule perforin, was shown to rely on normal expression of the NOS-2 gene.
- Published
- 2000
28. The Role of Integrins in Reproduction
- Author
-
Jeffery A. Bowen and Joan S. Hunt
- Subjects
Integrins ,media_common.quotation_subject ,Integrin ,Uterus ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Pregnancy ,Placenta ,medicine ,Animals ,Humans ,Embryo Implantation ,media_common ,Reproduction ,Placentation ,Embryo ,medicine.disease ,Cell biology ,medicine.anatomical_structure ,Fertilization ,Immunology ,biology.protein ,Female ,Function (biology) - Abstract
Fertilization, implantation, and placentation are dynamic cellular events that require not only synchrony between the maternal environment and the embryo, but also complex cell-to-cell communication. This communication involves integrins, a large family of proteins involved in the attachment, migration, invasion, and control of cellular function. Over the past decade, investigators have learned that integrins participate in multiple reproductive events including fertilization, implantation, and placentation in many species. This review will describe: (i) the expression of integrins on gametes and during the establishment and development of the placenta; (ii) regulatory pathways for controlling expression of integrins in the uterus and developing placenta; (iii) the function of integrins as determined by null-mutations; and (iv) reproductive dysfunction in women related to inappropriate integrin expression in the uterus and/or placenta.
- Published
- 2000
29. Unique epitopes of lactoferrin expressed in human cytotrophoblasts involved in immunologic reactions
- Author
-
W. Page Faulk, John A. McIntyre, Christian J. Thaler, Joan S. Hunt, and Carlos A. Labarrere
- Subjects
medicine.drug_class ,Inflammation ,Human leukocyte antigen ,Monoclonal antibody ,Epitope ,Amidohydrolases ,Epitopes ,Mice ,Pregnancy ,Placenta ,medicine ,Animals ,Humans ,Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase ,Trypsin ,Mice, Inbred BALB C ,biology ,Lactoferrin ,Macrophages ,Antibodies, Monoclonal ,Obstetrics and Gynecology ,HLA-DR Antigens ,Molecular biology ,Peptide Fragments ,Trophoblasts ,medicine.anatomical_structure ,embryonic structures ,biology.protein ,Immunohistochemistry ,Female ,Cytotrophoblasts ,Chorionic Villi ,medicine.symptom - Abstract
Objective: Lactoferrin is an iron-binding protein that has been implicated in protection against infections and allogeneic recognition reactions and in the control of cell growth. We studied the biochemical characteristics and expression of the unique lactoferrin epitopes (LF 1 ) in human placentas. Study Design: Immunohistologic studies of normal human term placentas were done by using monoclonal antibodies to LF 1 . Double-antibody experiments were done by using monoclonal antibodies to markers of inflammation (macrophages, human leukocyte antigen [HLA-DR]). LF 1 was studied immunochemically by using lactoferrin fragments produced by the reaction of lactoferrin with trypsin or N -glycanase. Results: Anti-LF 1 monoclonal antibodies reacted with most interstitial cytotrophoblasts in the basal plate and with villous cytotrophoblasts of some but not all chorionic villi. Cytotrophoblasts expressing LF 1 were associated with large numbers of HLA-DR–reactive macrophages. Anti-LF 1 monoclonal antibodies reacted with 2 distinct tryptic fragments of lactoferrin, and these reactivities were not affected by treatment with N -glycanase. Conclusion: Placental cytotrophoblasts express unique epitopes of lactoferrin (LF 1 ). Such expression is increased in the presence of activated macrophages. This expression could be an extraembryonic response to inflammation and maternal allogeneic recognition as an effort to protect trophoblastic cells. The LF 1 epitopes might represent conserved polypeptide epitopes on 2 homologous lobes of lactoferrin. (Am J Obstet Gynecol 1999;181:460-7.)
- Published
- 1999
30. Uterine Natural Killer Cells Are Targets for a Trophoblast Cell-Specific Cytokine, Prolactin-Like Protein A*
- Author
-
Bing Liu, Judith R. Head, Michael J. Soares, Heiner Müller, Joan S. Hunt, B. Anne Croy, and Guoli Dai
- Subjects
Cytotoxicity, Immunologic ,medicine.medical_specialty ,Recombinant Fusion Proteins ,medicine.medical_treatment ,Population ,chemical and pharmacologic phenomena ,Pregnancy Proteins ,Biology ,Mice ,Interleukin 21 ,Endocrinology ,Pregnancy ,immune system diseases ,Internal medicine ,medicine ,Animals ,education ,Cells, Cultured ,education.field_of_study ,Lymphokine-activated killer cell ,Uterus ,Decidua ,Trophoblast ,Alkaline Phosphatase ,Natural killer T cell ,Rats ,nervous system diseases ,Killer Cells, Natural ,medicine.anatomical_structure ,Cytokine ,Interleukin 12 ,Female ,lipids (amino acids, peptides, and proteins) - Abstract
PRL-like protein A (PLP-A) is a member of the PRL family expressed in trophoblast cells coincident with establishment of the chorioallantoic placenta. The purpose of this investigation was to identify targets for PLP-A. Using an alkaline phosphatase-tagging strategy, we show that PLP-A specifically interacts with a population of natural killer (NK) lymphocytes within the mesometrial compartment of decidua from pregnant and pseudopregnant rats. These observations are supported by the codistribution of PLP-A targets with cells expressing the rat NK cell surface marker, gp42, the absence of PLP-A binding in conceptuses from NK cell-deficient tgε26 mice, and the specific interaction of PLP-A with a rat NK cell line, RNK-16. We have further demonstrated that PLP-A effectively suppresses RNK-16 cell cytolytic activities. Our results provide evidence for a new paradigm of embryonic-maternal communication involving a PLP-A signaling pathway between trophoblast cells and uterine NK lymphocytes.
- Published
- 1999
31. TRAIL (Apo-2L) and TRAIL Receptors in Human Placentas: Implications for Immune Privilege
- Author
-
Teresa A. Phillips, Jian Ni, Guohua Pan, Steven M. Ruben, Ying-Fei Wei, Judith L. Pace, and Joan S. Hunt
- Subjects
Immunology ,Immunology and Allergy - Abstract
Mechanisms accounting for protection of the fetal semiallograft from maternal immune cells remain incompletely understood. In other contexts, interactions between TRAIL (TNF-related apoptosis-inducing ligand/Apo-2L) and its receptors kill activated lymphocytes. The purpose of this study was therefore to investigate the potential of the TRAIL/TRAIL-R system to protect the placenta against immune cell attack. Analysis by Northern blotting demonstrated mRNAs encoding TRAIL as well as the four TRAIL receptors (DR4, DR5, DcR1/TRID, DcR2/TRUNDD) in human placentas. Immunohistochemical experiments demonstrated that TRAIL protein is prominent in syncytiotrophoblast, an uninterrupted placental cell layer that is continuously exposed to maternal blood, as well as in macrophage-like placental mesenchymal cells (Hofbauer cells). Studies on cell lines representing trophoblasts (Jar, JEG-3 cells) and macrophages (U937, THP-1 cells) showed that both lineages contained TRAIL mRNA and that steady state levels of transcripts were increased 2- to 11-fold by IFN-γ. By contrast, cell lineage-specific differences were observed in expression of the TRAIL-R genes. Although all four lines contained mRNA encoding the apoptosis-inducing DR5 receptor, only trophoblast cells contained mRNA encoding the DcR1 decoy receptor and only macrophages contained DcR2 decoy receptor transcripts. DR4 mRNA was present only in THP-1 cells and was the only TRAIL-R transcript increased by IFN-γ. Cytotoxicity assays revealed that the two trophoblast cell lines were resistant, whereas the two macrophage lines were partially susceptible to killing by rTRAIL. Collectively, the results are consistent with a role for the TRAIL/TRAIL-R system in the establishment of placental immune privilege.
- Published
- 1999
32. Expression of Cell Adhesion Molecules in Murine Placentas and a Placental Cell Line1
- Author
-
Joan S. Hunt and Jeffery A. Bowen
- Subjects
Cell adhesion molecule ,Integrin ,Trophoblast ,Cell Biology ,General Medicine ,Adhesion ,Biology ,Molecular biology ,medicine.anatomical_structure ,Reproductive Medicine ,Cell culture ,Giant cell ,Placenta ,embryonic structures ,medicine ,biology.protein ,Cell adhesion ,reproductive and urinary physiology - Abstract
Integrins and vascular cell adhesion molecule-1 (VCAM-1) are required for normal placental development. In this study, integrin subunits a4, av, b1, and b3, and VCAM-1 were investigated for expression in uteroplacental units (gestation day [g.d.] 6 and 8) and placentas (g.d. 10, 12, 14, 16, and 18) of Swiss-Webster mice. All subunits and VCAM-1 mRNA (identified by reverse transcriptase polymerase chain reaction [RT-PCR]) and protein (detected by immunofluorescence) were present in all tissues throughout gestation. VCAM-1 was expressed strongly in the ectoplacental cone and trophoblast giant cells, a 4w as expressed strongly by trophoblast giant cells and moderately by spongiotrophoblast and labyrinthine trophoblast, and av was expressed more strongly in the spongiotrophoblast than in the labyrinthine zone. The b1 was more strongly expressed in the labyrinthine than the spongiotrophoblast zone, while b3 and VCAM-1 were essentially equal in the two zones. Trophoblastlike SM9‐1 cells were positive for all of the adhesion molecules when tested by RT-PCR and immunocytochemistry. Adhesion molecule expression in SM9‐1 cells was consistent with expression in the labyrinthine zone. Collectively, the results of this study demonstrate that murine placentas contain mRNA and protein for a4, av, b1, b3, and VCAM-1, and that expression is cell-specific. These results and the identification of an adhesion molecule-expressing trophoblastic cell line should facilitate future studies on the function of adhesion molecules in placental development.
- Published
- 1999
33. Apoptosis-inducing members of the tumor necrosis factor supergene family: Potential functions in placentae
- Author
-
Horst Bluethmann, Jeffery A. Bowen, Christine A. Rasmussen, Joan S. Hunt, and Teresa A. Phillips
- Subjects
Infertility ,Obstetrics and Gynecology ,Biology ,Bioinformatics ,medicine.disease ,Fas ligand ,Reproductive Medicine ,Apoptosis ,Immunology ,medicine ,Gene family ,Tumor necrosis factor alpha ,Receptor ,Function (biology) ,Developmental Biology ,Supergene - Abstract
Summary Apoptosis-inducing members of the TNF supergene family are clearly involved in local maternal-fetal immunological relationships as well as placental development and function. Although significant progress has been made in mapping out the temporal and spatial distribution of the best known members of this powerful gene family, TNFα, LTα and FasL, many newly reported ligands and receptors have yet to be examined. Furthermore, it is not yet clear how broad the range of functions of these cytokines may be or whether their death-dealing property is the most important. Nor is it certain whether each factor has an individual, specific role or whether overlapping functions will be the rule. Although developing a full understanding of the cytokines that comprise this family is a daunting task as greater and greater complexities are revealed, sorting out the expression, regulation and activities of each could yield major benefits. Infertility in one in six couples is now a widely quoted figure ( ISLAT Working Group, 1998 ) and it is likely that some portion of reproductive failure is due to inappropriate PCD and ACD mediated by members of the TNF supergene family.
- Published
- 1999
34. Immunobiology of Reproduction
- Author
-
Joan S. Hunt and Joan S. Hunt
- Subjects
- Reproduction--Immunological aspects--Congresse, Human reproduction--Immunological aspects--Con, Reproduction--immunology--congresses, Growth Substances--immunology--congresses
- Abstract
Over the last several decades, many biologists have been intrigued with the'immunological paradox'of pregnancy, where maternal and fetal tissues peaceably coexist despite their genetic differences. With the development of new insights on the interactive components of the immune system as well as the generation of powerful new molecular and cellular tools, singular progress has been made in understanding immunological events that culminate in successful pregnancy. This volume contains the proceedings for the first US symposium on the'Immunobiology of Reproduction'held August 26 - 29, 1993 in Boston, Massachusetts. Papers address hemotopoietic cells in reproductive tissues; growth factors/cytokines in the female reproductive tract and placenta; growth factor networks in pregnancy loss and cancer; placental expression of major histocompatibility complex and associated genes; experimental models of MHC gene expression; and immunological aspects of human infertility.
- Published
- 2012
35. Interferon-γ gene expression in cycling and pregnant mouse uterus: temporal aspects and cellular localization
- Author
-
J S Platt and Joan S. Hunt
- Subjects
medicine.medical_specialty ,Placenta ,medicine.medical_treatment ,Immunology ,Gene Expression ,In situ hybridization ,Biology ,Andrology ,Interferon-gamma ,Mice ,Paracrine signalling ,Estrus ,Pregnancy ,Interferon ,Internal medicine ,medicine ,Animals ,Immunology and Allergy ,Interferon gamma ,Autocrine signalling ,In Situ Hybridization ,reproductive and urinary physiology ,Cellular localization ,Uterus ,Trophoblast ,Cell Biology ,Immunohistochemistry ,medicine.anatomical_structure ,Endocrinology ,Cytokine ,Pregnancy, Animal ,Female ,medicine.drug - Abstract
Interferon-γ (IFN-γ) is a potent proinflammatory cytokine that modulates hematopoietic cell maturation, differentiation, activation, and apoptosis. To evaluate the postulate that locally produced IFN-γ could influence uterine hematopoietic cells, specific protein was detected by immunohistochemistry and messenger RNA (mRNA) was identified by in situ hybridization in cycling and pregnant mouse uteri. In cycling uteri, IFN-γ was limited to luminal and glandular epithelial cells during the estrus phase of the cycle. In pregnant uteri, IFN-γ was prominent at early (gestation day 6–10) and late (gestation day 18) stages. IFN-γ-producing cells identified by in situ hybridization included uterine epithelial cells, natural killer cells, macrophages, placental trophoblast cells, and cells in the degenerating metrial gland. Collectively, the data indicate that programming of immune and other cells via autocrine and paracrine pathways could be achieved by locally produced IFN-γ, and suggest that uteroplacental IFN-γ may be most influential during early and late stages of pregnancy. J. Leukoc. Biol. 64: 393–400; 1998.
- Published
- 1998
36. Differential expression and regulation of a human transgene, HLA-B27, in mouse placental and embryonic cell lines
- Author
-
Judith L. Pace, Joan S. Hunt, and Snigdha Banerjee
- Subjects
Embryology ,Somatic cell ,Placenta ,Mice, Transgenic ,Human leukocyte antigen ,Biology ,Cell Line ,Interferon-gamma ,Mice ,Antigen ,Pregnancy ,Genetics ,HLA-B Antigens ,medicine ,Animals ,Humans ,RNA, Messenger ,Transgenes ,Histocompatibility Antigen H-2D ,Molecular Biology ,HLA-B27 Antigen ,reproductive and urinary physiology ,Homozygote ,H-2 Antigens ,Gene Expression Regulation, Developmental ,Obstetrics and Gynecology ,Trophoblast ,Cell Biology ,Fibroblasts ,Embryo, Mammalian ,Embryonic stem cell ,Molecular biology ,Trophoblasts ,Mice, Inbred C57BL ,Reverse transcription polymerase chain reaction ,medicine.anatomical_structure ,Reproductive Medicine ,Cell culture ,embryonic structures ,Female ,Developmental Biology - Abstract
Unlike other somatic cells, human placental trophoblast cells do not express the highly polymorphic HLA-A and HLA-B human leukocyte major histocompatibility antigens that would stimulate maternal immunological rejection of the fetus. To investigate mechanisms underlying cell lineage-specific expression, cell lines were generated from homozygous matings of HLA-B27 transgenic mice. Trophoblast cell lines were generated from gestation day 10 placentas and fibroblasts were cultured from gestation day 13/14 embryos. Polymerase chain reaction (PCR) readily identified HLA-B DNA in transgenic trophoblastic cells but specific mRNA was of low abundance, being detectable by reverse transcriptase PCR but not by Northern blot hybridization. HLA-B-specific protein in/on the trophoblast cells was undetectable by cell enzyme-linked immunosorbent assay and the protein was not induced by exposing the trophoblastic cells to interferon-gamma (IFN-gamma). Restricted expression was specific for the HLA-B transgene and its antigen; IFN-gamma-inducible endogenous H-2Db class I antigens were detectable on the trophoblast cells. In contrast to the trophoblastic cells, HLA-B27 transgenic fibroblasts expressed IFN-gamma-inducible HLA class I antigens as well as H-2Db antigens. Thus, the mechanism(s) regulating expression of the polymorphic HLA-B antigen in trophoblastic cells is gene-specific, IFN-gamma-resistant and operative at the level of transcription or immediate post-transcription.
- Published
- 1998
37. Regulation of TNF-α Production in Activated Mouse Macrophages by Progesterone
- Author
-
Lance Miller and Joan S. Hunt
- Subjects
Immunology ,Immunology and Allergy - Abstract
The purpose of this study was to investigate the relationships between macrophage production of TNF-α and female hormones. Northern blot hybridization experiments showed that the female sex steroid hormone, progesterone, decreases steady state levels of TNF-α mRNA in LPS-activated mouse macrophages (RAW 264.7 and ANA-1 cells) in vitro. The production of intracellular and secreted TNF-α protein, as determined by ELISA, was decreased in both progesterone- and dexamethasone-treated, LPS-stimulated macrophages. Estrogen had no effect on expression of the TNF-α gene in mouse macrophages and did not alter progesterone-mediated suppression. Additional experiments conducted to investigate the mechanism of action of progesterone showed that this hormone, like dexamethasone, elevates steady state mRNA levels of IκBα and increases the levels of IκBα protein that are translocated from the cytoplasm to the nucleus. Thus, progesterone is a potent inhibitor of steady state levels TNF-α mRNA and TNF-α protein production in activated macrophages and may achieve this result through effects on an inhibitor of NF-κB.
- Published
- 1998
38. The tumor necrosis factor gene family in pregnancy
- Author
-
Christine A. Rasmussen and Joan S. Hunt
- Subjects
Genetically modified mouse ,Immune system ,Reproductive Medicine ,CD30 ,Cell culture ,Immunology ,Obstetrics and Gynecology ,Gene targeting ,Tumor necrosis factor alpha ,Biology ,Receptor ,Embryonic stem cell ,Developmental Biology - Abstract
Summary Even though many cytokines and growth factors first identified in the immune system are now known to participate in reproductive processes, investigators have been slow to recognize and evaluate the potential involvement of TNF family members. Yet recent studies illustrate the probability that several members of this family are important, particularly TNFα, LTα, CD30 and LTβ. The amn, gld and lpr mutant mouse strains, transgenic mice made deficient in TNF superfamily members and their receptors by gene targeting in embryonic stem cells as well cell lines developed from these mice should be extremely useful in learning how these potent, pleiotrophic cytokines contribute to successful pregnancy.
- Published
- 1998
39. Nonspecific immunity in the uterus and placenta
- Author
-
Joan S. Hunt, Lance Miller, and Wenjiang Chu
- Subjects
Fetus ,Pregnancy ,Innate immune system ,Obstetrics and Gynecology ,Biology ,medicine.disease ,Immune system ,medicine.anatomical_structure ,Reproductive Medicine ,Immune privilege ,Placenta ,Immunology ,medicine ,Tumor necrosis factor alpha ,Cell activation ,Developmental Biology - Abstract
Summary The studies described above clearly illustrate the principle that preventing the mother from rejecting her “foreign” fetus takes precedence over protecting the pregnancy from infection or invasion by foreign cells. In a recent commentary on the conflicting needs of pregnancy, Nahmias and Kourtis (1997) concluded that this causes no unacceptable risk to perpetuation of the species. Our experiments would support this conclusion; the pathways uncovered thus far are cleverly designed so that the negative effects are likely to be comparatively inconsequential. Regarding production of soluble HLA-G in macrophages, synthesis is linked to cell activation, a state wherein the phagocytic and degradative functions of the macrophages are improved. No conflict thus ensues; innate immunity provided by the macrophages would be enhanced rather than impaired by conditions associated with successful pregnancy. It will be extremely interesting to learn whether soluble HLA-G does stimulate programmed cell death in T lymphocytes and what concentrations of soluble HLA-G are required to inhibit NK cells. As for reduced production of NO and TNF, tight control is highly desirable; excessive production of either substance could interrupt pregnancy. It is therefore satisfying to find that progesterone, the dominant hormone of pregnancy, provides the control. Furthermore, the elegant dose-dependent system uncovered in our experiments, where only high concentrations of progesterone inhibit NO and TNF production, assures that synthesis of these two powerful molecules will be unimpaired in macrophages residing in tissues and organs distant from the progesterone-producing ovaries and placeta. As illustrated in the studies described above and recently summarized ( Hunt and Hutter, 1996 ), the mechanisms protecting the fetus from destruction by the maternal immune system are redundant and complex. Significant progress is being made in identifying and elucidating the specific features of each system. Concurrently, related issues such as the risks of chimerism in the mother and fetus and mechanisms underlying immune privilege are under intensive investigation ( Nelson, 1996 ; Runic et al., 1996 ; Green and Ware, 1997 ; Hunt et al., 1997b ).
- Published
- 1998
40. HLA Expression at the Maternal-Fetal Interface
- Author
-
Astrid Hammer, Gottfried Dohr, Joan S. Hunt, and Heinz Hutter
- Subjects
lcsh:Immunologic diseases. Allergy ,placenta ,HLA class l ,Genes, MHC Class I ,Human leukocyte antigen ,Biology ,immunology ,Immune system ,Antigen ,Pregnancy ,Placenta ,MHC class I ,medicine ,Humans ,Gene ,Maternal-Fetal Exchange ,Fetus ,Histocompatibility Antigens Class I ,Trophoblast ,trophoblast ,Trophoblasts ,medicine.anatomical_structure ,Immunology ,biology.protein ,Female ,MHC ,lcsh:RC581-607 ,Developmental Biology ,Research Article - Abstract
Pregancy in the human presents an “immunological paradox,” because of the unexpected willingness of mothers to accept genetically disparate tissues. The fact that the fetus can develop unharmed for nine months shows that protective mechanisms must exist to permit its survival. The conditions that permit the genetically dissimilar human fetus to evade rejection by its mother's immune system have been the subject of intense interest for several decades. As the placental cells, which are in contact with maternal blood or tissue, are devoid of HLA class II antigens, interest has focused on the expression of HLA class molecules. Recent developments in the constitutive, transcriptional, and translational expression of HLA class I molecules on anatomically and morphologically different subpopulations of trophoblast cells will form the basis of this short review.
- Published
- 1998
41. Expression of the Inducible Nitric Oxide Synthase Gene in Mouse Uterine Leukocytes and Potential Relationships with Uterine Function during Pregnancy1
- Author
-
Joan S. Hunt, B A Croy, Lance Miller, and Dianne Vassmer
- Subjects
Genetically modified mouse ,medicine.medical_specialty ,biology ,Uterus ,Trophoblast ,Cell Biology ,General Medicine ,Molecular biology ,Nitric oxide synthase ,Endocrinology ,medicine.anatomical_structure ,Reproductive Medicine ,Internal medicine ,Placenta ,embryonic structures ,Gene expression ,medicine ,biology.protein ,Metrial Gland ,Antibody - Abstract
Nitric oxide (NO), a potent and versatile free radical, is synthesized in leukocytes by the inducible form of NO synthase (iNOS). In this study, leukocytes in pregnant mouse uterus were investigated for expression of the iNOS gene. Inducible NOS mRNA, which was identified by reverse transcriptase polymerase chain reaction, was high relative to an invariant mRNA, glyceraldehyde-3-phosphate dehydrogenase, in midgestation uteri (gestation days [g.d.] 10, 12, and 14) but was low in late-gestation uteri (g.d. 16 and 18). Inducible NOS protein, identified immunohistochemically in paraformaldehydefixed uteri taken from g.d. 6 through 18 using rabbit antibodies generated to mouse carboxyl terminus iNOS peptides, was prominent in a few myometrial mast cells at early stages and was strongly expressed from g.d. 6 through g.d. 14 in myometrial macrophage-like cells. Inducible NOS protein was first detected in uterine (u) natural killer (NK) cells at g.d. 8. Signals peaked in this lineage at g.d. 10 and declined thereafter. Uterine leukocytes cultured in vitro expressed the iNOS gene; a hybridoma cell line derived from mouse uNK cells (GWM1-2) contained iNOS mRNA, and cells migrating from mouse metrial gland explants included iNOS+ leukocytes. Large, granular iNOS+ uNK cells were absent from the uteri of homologously mated pregnant TgE26 mice, an NK cell-deficient transgenic mouse strain, but immunoreactive iNOS was detectable in trophoblast, a cell lineage that did not contain immunoreactive iNOS in NK cell-competent Swiss-Webster mice. In TgE26 mothers gestating normal embryos, the same pattern was observed. Collectively, the results of this study demonstrate that iNOS is present in mouse uterine leukocytes including mast cells, macrophage-like cells, and uNK cells, and suggest that in the absence of uNK cells, the placenta synthesizes iNOS. These findings are consistent with the postulate that leukocyte NO contributes importantly to events associated with successful pregnancy that are likely to include relaxation of vascular smooth muscle.
- Published
- 1997
42. Soluble HLA-G isoforms: technical deficiencies lead to misinterpretations
- Author
-
Joan S. Hunt and Daniel E. Geraghty
- Subjects
Gene isoform ,Embryology ,Messenger RNA ,Cytotrophoblast ,medicine.drug_class ,Obstetrics and Gynecology ,Trophoblast ,Cell Biology ,Transfection ,Human leukocyte antigen ,Biology ,Monoclonal antibody ,Molecular biology ,medicine.anatomical_structure ,Syncytiotrophoblast ,Reproductive Medicine ,embryonic structures ,Genetics ,medicine ,Molecular Biology ,Developmental Biology - Abstract
The recent report authored by Blaschitz et al. (in press) would lead the reader to believe that human trophoblast cells cannot synthesize or secrete any of the soluble isoforms derived from splice variants of the HLA-G message. They propose that all of the soluble HLA-G presumed to be produced in trophoblast cells is, in fact, cleaved membrane HLA-G1. An interesting idea. Yet several published works have presented solid, well documented evidence that argues against this position. The original article describing the molecular structure of soluble HLA-G (Fujii et al., 1994) showed that soluble protein encoded by the intron 4-containing mRNA could be clearly distinguished from membrane HLA-G1 by isoelectric focusing (IEF). Further, a class I protein (reactive with W6/32) with an IEF pattern completely overlapping with soluble HLA-G from transfected cells was demonstrated in JEG-3 choriocarcinoma cells. Also in that study, PCR primers specific for soluble HLA-G-encoding mRNA were clearly described and used to confirm the protein work. In 2002, Solier and colleagues showed that soluble, intron-4 retaining HLA-G1 (now known as HLA-G5) mRNA is present in villous cytotrophoblast cells and that the HLA-G5 protein is secreted by the cells (Solier et al., 2002). In 2003, a team working in the Geraghty laboratory reported that membrane HLA-G1 is essentially absent in placentas, whereas HLA-G5 is abundant in both villous cytotrophoblast cells and syncytiotrophoblast (Ishitani et al., 2003). HLA-G1 was identified only in migrating extravillous cytotrophoblast cells using the highly specific o1G monoclonal antibody. Also in that study, the soluble HLA-G1 specific antibody 16G1 was used to isolate protein directly from term placentas. Bound peptide was isolated from this complex and shown to have a nearly identical profile to peptide isolated from membrane HLA-G1, which was also isolated from placentas. These peptides collectively matched very precisely the peptide binding motif previously described for HLA-G (some peptides were identical), and that work left no doubt as to the source of that peptide by N-terminal sequencing of the heavy chain (Lee et al., 1995). Taken together, these facts leave no doubt that 16G1 does bind HLA-G produced in vivo, and can only be reconciled with the conclusions made by Blaschitz study if one posits that 16G1 instead binds membrane HLA-G1. That conclusion in turn contradicts several other studies including some of the results presented in the Blaschitz et al. paper (Blaschitz et al., in press). At the next level of study alternative HLA-G protein forms were investigated. Newly generated, fully characterized, isoform-specific monoclonal antibodies developed in the Hunt laboratory demonstrated that villous cytotrophoblast cells contain HLA-G5 and HLA-G6 mRNA but express only HLA-G5 protein (Morales et al., 2003). By contrast, expression of HLA-G2/G6 protein was identified exclusively in extravillous cytotrophoblast cells. Despite this strong and consistent evidence put forth by three independent laboratories, it is always important to critically examine new evidence contradicting previously held conclusions. Taking the major features of the Blaschitz paper one at a time, let us look at the evidence they present to support their conclusions.
- Published
- 2005
43. Tumor Necrosis Factors: Pivotal Components of Pregnancy?1
- Author
-
Lance Miller, Joan S. Hunt, and Hua-Lin Chen
- Subjects
Messenger RNA ,medicine.medical_specialty ,Pregnancy ,Necrosis ,medicine.medical_treatment ,Embryogenesis ,Alpha (ethology) ,Cell Biology ,General Medicine ,Biology ,medicine.disease ,Endocrinology ,Cytokine ,Reproductive Medicine ,Internal medicine ,medicine ,Cancer research ,Gestation ,Tumor necrosis factor alpha ,medicine.symptom - Abstract
Tumor necrosis factor alpha is a major component of the infection-stimulated cytokine milieux associated with preterm labor and pregnancy termination. As a consequence, this potent factor is thought by some to have exclusively negative effects on the course of pregnancy. Yet in humans and other mammals, messenger RNA hybridizing with tumor necrosis factor alpha cRNA probes and protein detected by anti-tumor necrosis factor alpha have been identified in normal cycling and pregnant uteri, placentas, and embryos, consistent with an important role for tumor necrosis factor in female reproduction and embryonic development. Here, evidence for this intriguing dichotomy is presented, unresolved aspects are discussed, and potential roles for uteroplacental and embryonic tumor necrosis factor are proposed.
- Published
- 1996
44. Cytokine regulation of HLA-G expression in human trophoblast cell lines
- Author
-
Daniel E. Geraghty, Joan S. Hunt, and Yaping Yang
- Subjects
medicine.drug_class ,medicine.medical_treatment ,Immunology ,Human leukocyte antigen ,Biology ,Monoclonal antibody ,Cell Line ,Interferon-gamma ,HLA-C ,Antigen ,HLA Antigens ,Pregnancy ,HLA-G ,medicine ,Humans ,Immunology and Allergy ,RNA, Messenger ,HLA-G Antigens ,Tumor Necrosis Factor-alpha ,Histocompatibility Antigens Class I ,Obstetrics and Gynecology ,Trophoblast ,Flow Cytometry ,Molecular biology ,Trophoblasts ,medicine.anatomical_structure ,Cytokine ,Reproductive Medicine ,embryonic structures ,biology.protein ,Cytokines ,Female ,Antibody - Abstract
HLA class I genes are differentially expressed among subpopulations of cells in first trimester human placentas. In this study, HLA class I protein was detected in extravillous cytotrophoblast cells by immunohistochemistry using the monoclonal antibody W6/32. In the same trophoblast subpopulation, class Ib proteins were identified with two monoclonal antibodies, 87G (anti-HLA-G) and 131 ( anti-HLA- A G ) and class Ia protein was detected with the monoclonal antibody, 4E ( anti-HLA- B C ). All of the antibodies also identified antigens on the human trophoblast-derived choriocarcinoma cell line, JEG-3. Therfore, the JEG-3 cells were used as a model system to study cytokine regulation of HLA-G in trophoblast cells. Northern blot hybridization studies showed that interferons (IFN-α, IFN-β, IFN-γ) and tumor necrosis factor-α (TNF-α) modestly enhanced steady state levels of HLA-G mRNA. Yet analysis of HLA-G protein by immunocytochemistry and flow cytometry failed to identify any changes in intracellular or membrane expression of HLA-G protein following cytokine treatment. Resistance to upregulation of HLA class I antigens was not a general feature of JEG-3 cells; IFNs enhanced expression of HLA- B C as well as HLA class I light chain, β2-microglobulin. HLA null Jar choriocarcinoma cells did not contain HLA-G mRNA or antigen and exposure to cytokines had no effect on HLA-G. The results of this study are consistent with the postulate that trophoblast cell expression of HLA-G is stringently regulated and is controlled in part by post-transcriptional mechanisms.
- Published
- 1995
45. Tumor Necrosis Factor- in the Female Reproductive Tract
- Author
-
Katherine F. Roby, Verda J. Hunter, Joan S. Hunt, and Paul F. Terranova
- Subjects
medicine.medical_specialty ,Placenta ,medicine.medical_treatment ,Biology ,Receptors, Tumor Necrosis Factor ,General Biochemistry, Genetics and Molecular Biology ,Paracrine signalling ,Pregnancy ,Internal medicine ,medicine ,Animals ,Humans ,Autocrine signalling ,Receptor ,Regulation of gene expression ,Tumor Necrosis Factor-alpha ,Reproduction ,Ovary ,Uterus ,Embryo ,Genitalia, Female ,Embryo, Mammalian ,Endocrinology ,Cytokine ,Gene Expression Regulation ,Female ,Tumor necrosis factor alpha ,Hormone - Abstract
Tumor necrosis factor-alpha (TNF), originally identified as an inflammation-associated cytokine, is synthesized throughout the female reproductive tract as well as in placentas and embryos. Development, female sex steroid hormones, and lipopolysaccharide influence expression of this gene. The functions of TNF may be determined in part by differential expression of the two species of TNF receptors, both of which seem to be regulated by female sex steroid hormones. Evidence has accumulated that supports a role for this potent, pleiotropic cytokine in autocrine and paracrine processes central to reproduction, including gamete and follicle development, steroidogenesis, uterine cyclicity, placental differentiation, development of the embryo, and parturition.
- Published
- 1995
46. Expression and localization of messenger RNA for tumor necrosis factor receptor (TNF-R) I and TNF-RII in pregnant mouse uterus and placenta
- Author
-
Paul F. Terranova, Nihay Laham, Joan S. Hunt, Hendrick Kröning, and Katherine F. Roby
- Subjects
medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Uterus ,Trophoblast ,In situ hybridization ,Biology ,Endometrium ,Endocrinology ,medicine.anatomical_structure ,Internal medicine ,Placenta ,medicine ,Decidual cells ,Tumor necrosis factor alpha ,Northern blot - Abstract
The temporal and cell-specific localization of tumor necrosis factor (TNF) receptor mRNAs in the uterus and placenta during pregnancy in the mouse was investigated. Messenger RNA for TNF and the TNF receptors (TNF-RI, p55/p60 and TNF-RII, p75/p80) was assessed by northern blot andin situ hybridization. TNF, TNF-RI and TNF-RII specific transcripts were present on days 7 through 18 of pregnancy. Relative concentrations of TNF mRNA decreased from days 7 to 18 with levels being higher in the uterus than the placenta. In contrast TNF-RI mRNA levels were constant throughout gestation and no differences were seen between steady state levels in the uterus and placenta. Two transcripts for TNF-RII (3.6 and 4.5 kb) were identified in all tissues. Steady state levels of TNF-RII mRNA increased throughout gestation and levels were higher in the placenta than in the uterus. On day 9 of gestation, TNF-RI and TNF-RII mRNAs were localized to undecidualized endometrium, mesometrial decidual cells, and the developing placenta. In addition, muscle cells contained TNF-RI but not TNF-RII mRNA. By day 15 of gestation, TNF-RI and TNF-RII transcripts were primarily localized to the uterine epithelium and trophoblast giant cells and spongiotrophoblast cells in the placenta. The results of these studies reveal the uterine and placental cell-specific expression of TNF receptor mRNAs during pregnancy in the mouse and provide insight into the cellular targets of TNF action.
- Published
- 1995
47. Myometrial Tumor Necrosis Factor α: Cellular Localization and Regulation by Estradiol and Progesterone in the Mouse1
- Author
-
Katherine F. Roby and Joan S. Hunt
- Subjects
Estrous cycle ,medicine.medical_specialty ,medicine.medical_treatment ,Immunocytochemistry ,Myometrium ,Cell Biology ,General Medicine ,Biology ,Mast cell ,Endocrinology ,Cytokine ,medicine.anatomical_structure ,Reproductive Medicine ,Internal medicine ,medicine ,Tumor necrosis factor alpha ,Cellular localization ,Immunostaining - Abstract
The expression of tumor necrosis factor α (TNF) protein in mouse myometrial cells on each day of the estrous cycle and after ovariectomy, with and without replacement of estradiol (E 2 ), progesterone (P 4 ), or E 2 +P 4 , was investigated. TNF protein was assessed by immunocytochemistry. In addition, the numbers of mast cells and the numbers of TNF-containing mast cells were determined under each condition. The total number of mast cells fluctuated during the estrous cycle; and at each stage, essentially 100% of the mast cells exhibited TNF immunoreactivity. In contrast, after ovariectomy and after ovariectomy with E 2 replacement, only ∼50% of the mast cells contained immunoreactive TNF. P 4 treatment resulted in further decline in mast cell TNF whereby after 24 h of P 4 , only 3% of the mast cells were TNF-positive and by 72 h of treatment, no TNF-positive mast cells could be detected. E 2 and P 4 in combination increased the total number of mast cells, and in a pattern reminiscent of normal cycling myometrium, over 90% of the mast cells exhibited TNF immunoreactivity after 24 h of treatment. TNF protein was detectable in muscle cells throughout the estrous cycle, with weak immunostaining observed on proestrus and more intense immunostaining on estrus, diestrus-I, and diestrus-II. After ovariectomy, only light immunostaining was observed in the muscle cells. Immunoreactive TNF was detected in muscle cells after each type of steroid treatment. E 2 and E 2 +P 4 treatments resulted in a biphasic pattern of immunoreactive TNF expression. Immunostaining intensity was increased at 6 h after treatment, decreased after 24 h, and increased again after 72 h. In contrast, the greatest intensity of TNF immunostaining was observed 1 h after P 4 administration and the staining intensity decreased thereafter. These studies reveal 1) the cellular localization of TNF protein in the mouse myometrium, 2) the identification of the mast cell as a major source of uterine TNF, and 2) regulation of myometrial TNF by E 2 and P 4
- Published
- 1995
48. Immunologically Relevant Cells in the Uterus1
- Author
-
Joan S. Hunt
- Subjects
Lymphocyte ,Cell ,Uterus ,Cell Biology ,General Medicine ,Biology ,Natural killer cell ,Cell biology ,Haematopoiesis ,medicine.anatomical_structure ,Reproductive Medicine ,Immunology ,medicine ,Interleukin 12 ,Macrophage ,Cell activation - Abstract
Macrophages and a special subset of lymphocyte natural killer (NK) cells populate the uteri of cycling humans, mice, and rats. After implantation, major changes take place that have important functional implications. The macrophages and NK cells increase in number, are redistributed into specific uterine compartments, and exhibit markers consistent with cell activation. Activation enhances macrophage and NK cell production of a wide range of pleiotropic, multifunctional polypeptide growth factors, reactive oxygen intermediates, and bioactive lipids. Thus, activated uterine hematopoietic cells are equipped to perform certain immunological and nonimmunological functions within their microenvironments that could have major influences on the course of pregnancy.
- Published
- 1994
49. Normal Distribution of Tumor Necrosis Factor-α Messenger Ribonucleic Acid and Protein in the Uteri, Placentas, and Embryos of Osteopetrotic (op/op) Mice Lacking Colony-Stimulating Factor-11
- Author
-
Hua-Lin Chen, Xiao-Ling Hu, Joan S. Hunt, and Jeffrey W. Pollard
- Subjects
Macrophage colony-stimulating factor ,medicine.medical_specialty ,medicine.medical_treatment ,Uterus ,Cell Biology ,General Medicine ,In situ hybridization ,Biology ,Andrology ,medicine.anatomical_structure ,Endocrinology ,Cytokine ,Reproductive Medicine ,Internal medicine ,Gene expression ,medicine ,Decidual cells ,Tumor necrosis factor alpha ,Northern blot - Abstract
In order to map mouse tumor necrosis factor-alpha (TNF) gene expression in detail and to determine whether transcription or translation of the TNF gene is regulated by uterine colony stimulating factor-1 (CSF-1), preimplantation embryos, oviducts, uteri, and uteroplacental units were studied in various strains of mice. These included homozygous osteopetrotic (op/op) female mice, which completely lack CSF-1, and heterozygous (+/op) females, which have normal levels of CSF-1. TNF mRNA was identified in all samples except preimplantation embryos by use of Northern blot hybridization or reverse transcriptase polymerase chain reaction. In situ hybridization and immunocytochemical experiments showed that the TNF gene was expressed in mouse oviduct and uterine epithelial cells, decidual cells, macrophage-like cells, placental trophoblast, and embryos. Despite an absence of CSF-1, TNF gene expression in the uteri, placentas, and embryos of op/op mothers did not differ in any major respect from expression in +/op or other strains of mice. The results of this study therefore indicate that the TNF gene is transcribed and translated in an ordered sequence through mouse gestation, and that maternal CSF-1 is not essential to expression of this cytokine gene. Collectively, these findings are consistent with a major role for TNF in mouse reproduction and development and with a potential compensatory function for this potent polypeptide factor in CSF-1 deficiency.
- Published
- 1993
50. Expression of tumor necrosis factor-alpha in mouse spermatogenic cells
- Author
-
Paul F. Terranova, G. C. Enders, Swapan K. De, Joan S. Hunt, Judith L. Pace, and Hua-Lin Chen
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,Gene Expression ,In situ hybridization ,Spermatocyte ,Biology ,Mice ,Endocrinology ,Internal medicine ,Testis ,medicine ,Animals ,RNA, Messenger ,Northern blot ,Cells, Cultured ,In Situ Hybridization ,Mice, Inbred ICR ,Sertoli Cells ,Spermatid ,Tumor Necrosis Factor-alpha ,urogenital system ,Leydig Cells ,Riboprobe ,Blotting, Northern ,Sertoli cell ,Spermatids ,Spermatozoa ,Blot ,medicine.anatomical_structure ,Liver ,Culture Media, Conditioned ,Spermatogenesis - Abstract
Enriched fractions of spermatogenic cells were isolated by unit gravity sedimentation and analyzed both for the presence of secreted tumor necrosis factor-alpha (TNF alpha) in vitro by bioassay and for the presence of TNF alpha mRNA by Northern blot analysis. Small quantities of bioactive TNF alpha were consistently detected in medium conditioned by round spermatid fractions. Both pachytene spermatocyte and round spermatid fractions contained RNA that hybridized with murine cDNA probes for TNF alpha, with pachytene spermatocytes containing a normal 1.9-kilobase (kb) transcript, while round spermatids contained principally an approximately 2.8-kb transcript. Both the normal size transcript and the larger haploid-specific transcript were enriched when total RNA from pachytene spermatocyte and round spermatid fractions was passed through an oligo(dT) column. The normal 1.9-kb transcript within pachytene spermatocytes could be induced by exposing the spermatogenic cells to lipopolysaccharides in vitro, yet the approximately 2.8-kb transcript within round spermatids appeared uninduced by LPS treatment. In situ hybridization for the TNF alpha message by using digoxigenin label antisense TNF alpha riboprobe labeled pachytene spermatocytes, round spermatids, and presumptive interstitial macrophages. Spermatogonia and elongating spermatids as well as other interstitial cells were unlabeled or very lightly labeled. Hybridization of 16-day-old prepuberal testis resulted in the labeling of spermatocytes and presumptive interstitial macrophages. RNA from Sertoli cells, but not pachytene spermatocytes or round spermatids, hybridized with human TNF alpha receptor p60 probe in Northern blot analysis. These results are consistent with the working hypothesis that spermatids release TNF alpha, which is detected by Sertoli cells and may serve as a paracrine factor, regulating an as yet unidentified process in spermatogenesis.
- Published
- 1993
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