Thematische Bibliographien / T cells Identification

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Auswahl der wissenschaftlichen Literatur zum Thema „T cells Identification“

Autor: Grafiati

Veröffentlicht am 4. Juni 2021

Zuletzt aktualisiert am 29. Januar 2023

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Zeitschriftenartikel zum Thema "T cells Identification"

Aerts, Nicolaas E., Evelyne J. Dombrecht, Didier G. Ebo, Chris H. Bridts, Wim J. Stevens und Luc S. De Clerck. „Activated T cells complicate the identification of regulatory T cells in rheumatoid arthritis“. Cellular Immunology 251, Nr. 2 (2008): 109–15. http://dx.doi.org/10.1016/j.cellimm.2008.04.008.

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Graca, Luis, Stephen P. Cobbold und Herman Waldmann. „Identification of Regulatory T Cells in Tolerated Allografts“. Journal of Experimental Medicine 195, Nr. 12 (10.06.2002): 1641–46. http://dx.doi.org/10.1084/jem.20012097.

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Induction of transplantation tolerance with certain therapeutic nondepleting monoclonal antibodies can lead to a robust state of peripheral “dominant” tolerance. Regulatory CD4+ T cells, which mediate this form of “dominant” tolerance, can be isolated from spleens of tolerant animals. To determine whether there were any extra-lymphoid sites that might harbor regulatory T cells we sought their presence in tolerated skin allografts and in normal skin. When tolerated skin grafts are retransplanted onto T cell–depleted hosts, graft-infiltrating T cells exit the graft and recolonize the new host. These colonizing T cells can be shown to contain members with regulatory function, as they can prevent nontolerant lymphocytes from rejecting fresh skin allografts, without hindrance of rejection of third party skin. Our results suggest that T cell suppression of graft rejection is an active process that operates beyond secondary lymphoid tissue, and involves the persistent presence of regulatory T cells at the site of the tolerated transplant.

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Souter, M. N., C. V. Nguyen-Robertson, F. J. Ross, S. J. J. Reddiex, J. Waddington, I. Van Rhijn, S. B. G. Eckle et al. „Identification and characterization of CD1-restricted T cells“. Journal of Immunology 196, Nr. 1_Supplement (01.05.2016): 206.6. http://dx.doi.org/10.4049/jimmunol.196.supp.206.6.

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Abstract Most studies of T cells have focused on those that respond to foreign peptides. However, other specialized populations of T cells exist that recognize lipid antigens and make up a substantial component of the human immune system. These lipid reactive T cells recognize antigens presented by antigen presentation molecules from the CD1 family. Four CD1 molecules exist (CD1a, CD1b, CD1c and CD1d), and each is capable of presenting a unique repertoire of lipids antigens to T cells. Much of what we have learned about lipid reactive T cells stems from studies of CD1d restricted NKT cells as these are present is both mice and humans and can be detected using CD1d/α-GalCer tetramers. In contrast, our understanding of the biology of CD1a, CD1b, CD1c restricted T cells is relatively limited. However, the recent generation of CD1a, CD1b and CD1c tetramers is helping with the identification and characterisation of these CD1-restricted T cells. We have produced CD1 tetramers loaded with mammalian self-lipids or lipid antigens from Mycobacterium tuberculosis. In conjunction, with a tetramer-based enrichment method, we have successfully identified both autoreactive and microbial lipid antigen specific T cells from healthy human blood. We reveal the phenotypic characteristics of these CD1-restricted T cells and used CD1 mutagenesis to provide new insight into TCR recognition of CD1-lipid antigen complexes. Collectively, these studies will serve as a basis for future studies of lipid reactive T cells in health and disease.

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Illes, Z., T. Kondo, K. Yokoyama, T. Ohashi, T. Tabira und T. Yamamura. „Identification of autoimmune T-cells among in vivo expanded CD25+ T-cells in multiple sclerosis“. Journal of Neuroimmunology 90, Nr. 1 (September 1998): 73. http://dx.doi.org/10.1016/s0165-5728(98)91617-4.

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Illés, Zsolt, Takayuki Kondo, Kazumasa Yokoyama, Takashi Ohashi, Takeshi Tabira und Takashi Yamamura. „Identification of Autoimmune T Cells Among In Vivo Expanded CD25+ T Cells in Multiple Sclerosis“. Journal of Immunology 162, Nr. 3 (01.02.1999): 1811–17. http://dx.doi.org/10.4049/jimmunol.162.3.1811.

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Abstract Although clonal expansion of autoimmune T cells has been reported in multiple sclerosis (MS), very limited information is available on specificities, clonal size, or activation state of the expanded clones. Here we address the issue of clonal expansion by using a novel technique demonstrating clonotypes defined by single-strand conformation polymorphism of TCR β-chain cDNAs. Examination of activated T cells (CD3+CD25+) isolated from the peripheral blood of MS revealed limited numbers (20∼82) of expanded clones defined by single-strand conformation polymorphism clonotype. To estimate the Ag specificities of dominant clonotypes in the activated T cells, these samples were examined in parallel with Th1 T cell clones specific for myelin basic protein or proteolipid protein (PLP) derived from the same patients. Analysis of two patients demonstrated that the dominant clonotypes would contain those specific for myelin basic protein or PLP. Although the majority of the clonotypes could be detected only transiently, a PLP95–116-specific clonotype was found to persist for over 1 yr. Thus, single-strand conformation polymorphism clonotype analysis allows us to monitor the kinetics of given T cell clones in vivo and could provide useful information for designing clonotype (Id)-specific manipulation of human diseases such as MS.

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Baseggio, L., F. Berger, D. Morel, M.-h. Delfau-Larue, G. Goedert, G. Salles, J.-p. Magaud und P. Felman. „Identification of circulating CD10 positive T cells in angioimmunoblastic T-cell lymphoma“. Leukemia 20, Nr. 2 (08.12.2005): 296–303. http://dx.doi.org/10.1038/sj.leu.2404013.

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Azimi, Maryam, Saeed Aslani, Sahar Mortezagholi, Amir Salek, Mohammad Reza Javan, Alireza Rezaiemanesh, Mojgan Ghaedi, Mehrdad Gholamzad und Eisa Salehi. „Identification, Isolation, and Functional Assay of Regulatory T Cells“. Immunological Investigations 45, Nr. 7 (15.07.2016): 584–602. http://dx.doi.org/10.1080/08820139.2016.1193869.

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Kobayashi, Hiroya, und Esteban Celis. „Peptide epitope identification for tumor-reactive CD4 T cells“. Current Opinion in Immunology 20, Nr. 2 (April 2008): 221–27. http://dx.doi.org/10.1016/j.coi.2008.04.011.

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Gerli, Roberto, Giuseppe Nocentini, Alessia Alunno, Elena Bartoloni Bocci, Rodolfo Bianchini, Onelia Bistoni und Carlo Riccardi. „Identification of regulatory T cells in systemic lupus erythematosus“. Autoimmunity Reviews 8, Nr. 5 (März 2009): 426–30. http://dx.doi.org/10.1016/j.autrev.2009.01.004.

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Cardarelli, P. M., und M. D. Pierschbacher. „Identification of fibronectin receptors on T lymphocytes.“ Journal of Cell Biology 105, Nr. 1 (01.07.1987): 499–506. http://dx.doi.org/10.1083/jcb.105.1.499.

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We report the identification of fibronectin receptors on thymocytes and T lymphoma cells. Affinity chromatography of extracts of the T cell lymphoma, WR16.1, on a fibronectin-Sepharose column combined with specific elution using a synthetic peptide containing the cell attachment-promoting sequence, arginine-glycine-aspartic acid, yielded two polypeptide components having apparent molecular masses of approximately 160 kD reduced and 175 and 150 kD nonreduced. Immunoprecipitations from surface-iodinated WR16.1 cells or fibronectin-adherent thymocytes using a rabbit antiserum raised against the fibronectin receptor that is present on human fibroblasts revealed, in each case, the same two radiolabeled components. In contrast, immunoprecipitation from fibronectin-nonadherent T lymphoma cells, designated WR2.3, revealed the presence of only the smaller subunit. Although the lymphocyte receptor and the fibronectin receptor identified on fibroblasts share immunologic determinants, they differ in that the molecular mass of the lymphocyte protein is larger. Moreover, trypsinization of either thymocytes or the WR16.1 T lymphoma cells resulted in a subsequent loss of their ability to adhere to fibronectin-coated substrates and a reduction in the electrophoretic mobility of each of the polypeptide chains of the fibronectin receptor present on their surfaces. These changes, however, were not observed with normal rat kidney fibroblasts or mouse 3T3 fibroblasts in response to trypsinization. The data establish the existence on normal lymphocytes of fibronectin receptors that are quite similar to those found on fibroblasts. The possible function of this molecule on thymocytes is discussed.

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Dissertationen zum Thema "T cells Identification"

Ye, Song Cheung H. Tak. „Identification of a thymic extracellular matrix protein that promotes strong thymocyte adhesion“. Normal, Ill. Illinois State University, 1990. http://wwwlib.umi.com/cr/ilstu/fullcit?p9115234.

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Thesis (Ph. D.)--Illinois State University, 1990.
Title from title page screen, viewed December 2, 2005. Dissertation Committee: H. Tak Cheung (chair), Herman Brockman, Harry Huizinga, Anthony Otsuka, Brian Wilkinson. Includes bibliographical references (leaves 116-127) and abstract. Also available in print.

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Wang, Min-Guang Cheung H. Tak. „Identification of an extracellular matrix epitope involved in T cell adhesion“. Normal, Ill. Illinois State University, 1992. http://wwwlib.umi.com/cr/ilstu/fullcit?p9311292.

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Thesis (Ph. D.)--Illinois State University, 1992.
Title from title page screen, viewed February 7, 2006. Dissertation Committee: H. Tak Cheung (chair), Mathew J. Nadakavukaren, Alan J. Katz, Brian J. Wilkinson, Lynne A. Lucher. Includes bibliographical references (leaves 103-114) and abstract. Also available in print.

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Garefalaki, Anna. „Identification of regulatory regions that determine expression of murine CD8 locus“. Thesis, University College London (University of London), 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.250198.

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Robinson, Jonathan Matthew. „Identification of tumour-specific T-cells in colorectal cancer patients“. Thesis, University of Leeds, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.485913.

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The ability to induce an immune response to a tumour is an attractive approach to cancer treatment, particularly when used in combination with surgery and chemotherapy or radiotherapy. In theory the use of such immunological techniques should be possible to help treat colorec~ cancer. Immunotherapy has shown encouraging results in the treatment of other cancers, namely malignant melanoma and renal cell cÃ‚Â¥cinoma: The ability of these techniques to aid the treatment of advanced colorectal cancer has so far proved disappointing, although some encouragement has been observed. It has recently become possible to identify CD8+ T-cells specific for particular antigens using MHC class I tetramer staining. Carcinoembryonic antigen (CEA) is one such tumo~ antigen that is normally only expressed during embryonic development and not by adult tissues. However, in patients with colorectal cancer, CEA is over expressed and is therefore considered to be a tumour-specific antigen (TAA). T-cells specific for an epitope of CEA (CAP-I) can be identified using CAP-I loaded MHC class I tetramer molecules. The aim of this project was to assess the number and frequency of CEA specific CD8+ T-cells in colorectal cancer patients using this tetramer staining'protocol; in particular during the different treatments they receive prior to surgery, namely radiotherapy and ' chemotherapy. Previous methods to study these 'cells required large volumes of blood not suitable for patient studies. A novel and enhanced technique for the generation of antigen specific T-cells was employed which only required small volumes of blood . ideal for repeated patient testing. Cells recovered from the peripheral blood of healthy controls were cultured and observed for the proliferation of CAP-I specific T-cells, which were detected by tetramer staining, replicating the results of previous studies.

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Hansson, Johan. „Activation and differentiation of cytotoxic T lymphocytes identification of district CTL subsets in the rat /“. Lund : Dept. of Tumor Immunology, the Wallenberg Laboratory, 1994. http://catalog.hathitrust.org/api/volumes/oclc/39158589.html.

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Lam, Eric M. „IDENTIFICATION OF CYCLIN-DEPENDENT KINASE 5 IN T CELLS AND ITS ROLE IN REGULATING T CELL FUNCTION AND DIFFERENTIATION“. Case Western Reserve University School of Graduate Studies / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=case1422014852.

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Ulaganathan, Vijay Kumar. „Gene Expression Profiling of Encephalitogenic CD4+ T cells: Identification of Genes Controlling Migration of Effector T cells into the CNS“. Diss., lmu, 2010. http://nbn-resolving.de/urn:nbn:de:bvb:19-122549.

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Bosco, Anthony. „Identification of novel genes associated with allergen-driven T cell activation in human atopics /“. Connect to this title, 2006. http://theses.library.uwa.edu.au/adt-WU2007.0023.

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Pandey, Shubham. „Identification of Interleukin 4 - CXCL12 supportive loop in follicular lymphoma“. Thesis, Rennes 1, 2016. http://www.theses.fr/2016REN1B031/document.

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Le lymphome folliculaire (FL) est le lymphome B indolent le plus fréquent. Outre des altérations géniques récurrentes, le micro-environnement tumoral, et notamment les cellules stromales lymphoides,joue un rôle majeur dans le développement de ce cancer. Cependant, la caractérisation in-situ des cellules stromales lymphoïdes chez l'homme tout comme les facteurs menant à la polarisation du stroma en un stroma protumoral ont été peu étudiés. Dans cette thèse, nous avons montré, que les cellules stromales présentes dans les ganglions et la moelle osseuse envahis des patients atteints de FL surexpriment fortement la chimiokine CXCL12. Nous avons ensuite tenté de comprendre les mécanismes responsables de cette induction. Alors que les cellules B tumorales induisent une surexpression de la chimiokine CCL2 dans les cellules stromales de façon dépendante de leur synthèse de TNF, elles ne contribuent pas à l'induction de CXCL12. A l'inverse, le principal compartiment TCD4 impliqué dans la croissance tumorale du FL, les cellules T follicular helper (TFH), augmentent l'expression de CXCL12 dans les cellules stromales. Le taux d'IL-4, la principale cytokine produite par les TFH de FL, est d'ailleurs corrélé à celui de CXCL12 au sein de ma niche tumorale du FL. De plus, à l’aide d'un modèle de différenciation en stroma lymphoide, nous avons démontré que l’IL4 induit l’expression de CXCL12 par les cellules stromale in vitro. Cette production est augmentée quand les cellules stromales sont déjà engagées vers la voie de différentiation lymphoide par un traitement TNF/LT qui favorise l'activation de STAT6 par l'IL-4. Nous avons validé ces résultats dans un modèle de formation d'organe lymphoide ectopique chez la souris. Enfin, CXCL12 induit la migration et l'adhésion au stroma des B de FL via l'activation de cascades de signalisations qui peuvent être abrogées par l'utilisation d'un inhibiteur de Btk utilisé en clinique, l'Ibrutinib. Ces résultats sont en faveur de l'intérêt de considérer la boucle IL-4/CXCL12 pour développer de nouvelles stratégies thérapeutiques dans cette pathologie constamment fatale
Follicular lymphoma (FL) is the most frequent indolent B-cell lymphoma. Beside recurrent genetic alterations, tumor microenvironment, including lymphoid stromal cells, has been shown to play a key role in FL development. However, in situ characterization of lymphoid stromal cells is still lacking in humans and there are very few studies focusing on the factors that could lead to stroma polarization in normal and pathological context. In this thesis, we showed first that in FL, lymph node (LN) and bone marrow (BM) infiltrating stromal cells highly express the chemokine CXCL12. We next focused on the mechanisms underlying this upregulation. Interestingly, whereas malignant FL B cells induced overexpression of CCL2 in stromal cells in a TNF-dependent manner, they did not contribute to CXCL12 induction. Conversely, FL-infiltrating follicular helper T cells (FL-TFH), the key FL-supportive T-cell subset could trigger CXCL12 expression in stromal cells. IL-4 is the main FL-TFH-derived cytokine and showed a positive correlation with CXCL12 expression inside FL cell niches. Moreover, based on our in vitro lymphoid stroma differentiation model, we demonstrated that IL-4 promoted CXCL12 expression in stromal cells, together with a phenotype close to that identified in situ within FL cell niche. Such IL4 dependent CXCL12 regulation is more pronounced in stromal cells already committed towards lymphoid stromal cells by a prestimulation by TNF/LT in association with an increased STAT6 activation. These data were validated in a model of ectopic lymphoid organ formation in mice. Finally, CXCL12 induced FL B-cell migration, and adhesion to stromal cells through the activation of a signaling pathway that could be abrogated by the Btk inhibitor Ibrutinib. These data argue for considering IL-4/CXCL12 axis as a potential therapeutic target to disrupt FL protective cell niche in this still fatal malignancy

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Bosco, Anthony. „Identification of novel genes associated with allergen-driven T cell activation in human atopics“. University of Western Australia. School of Paediatrics and Child Health, 2007. http://theses.library.uwa.edu.au/adt-WU2007.0023.

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[ Truncated abstract ] Atopic diseases such as asthma are thought to be driven to a significant extent by T helper memory cells which are programmed to respond in a harmful way to environmental allergens (e.g. house dust mite). Previous studies in humans and in animal models have established that activation of TH2 cytokine genes in T memory responses to allergens is central to the disease process. However, only a subset of atopics harbouring a TH2-memory response phenotype manifests clinical symptoms of disease. Moreover, clinical trials with TH2 antagonists in atopic patients have proven disappointing, suggesting underlying complexities in disease pathogenesis which escape regulation via these approaches. It was thus hypothesised that additional genes involved in the activation program of allergen-specific T memory cells which are central to disease pathogenesis remain unidentified. The aim of the current study was to identify such novel genes by applying microarray technology to survey genome-wide expression patterns in an in vitro model of allergen-driven human T cell activation. In contrast to previous human microarray studies in this area focusing on mitogen activated T cell lines and clones, the current study avoided the use of strong activation stimuli which have the potential to distort patterns of gene expression, and reports for the first time the findings of microarray analysis of house dust mite allergen-driven acute gene activation in recirculating T memory cells harvested from the peripheral blood of human atopics. ... Finally, methodology was established to investigate the function of the novel atopy-associated genes. In loss-of-function experiments, expression of DACT1 and CAMK2D was silenced in primary T cell responses driven by bacterial superantigens, a model system for studying T cell responses under conditions which mimic antigen-specific activation. Whilst silencing DACT1 and CAMK2D expression did not influence classical readouts of T cell function including proliferation and cytokine production, microarray profiling was employed to identify putative downstream transcriptional targets of each gene. The experimental strategy and optimised methodology presented herein can now be employed to investigate the molecular circuitry linking the novel atopy-associated genes to the T cell activation process. In conclusion, several novel genes associated with allergen-driven T memory responses in atopics have been first described in this thesis and represent logical candidates for more detailed immunological and genetic studies related to the pathogenesis of atopic diseases.

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Bücher zum Thema "T cells Identification"

Sjoerd Henricus van der Burg. Identification and evaluation of cytotoxic T-cell epitopes in HIV-1 and tumour-associated proteins. [Leiden: University of Leiden, 1998.

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Loreto, Michael Peter. Identification and characterization of Src-like adaptor protein-2, a novel hematopoietic-specific adaptor protein and inhibitor of T-cell receptor signaling. Ottawa: National Library of Canada, 2002.

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Identification and characterization of murine TCR [gamma] [delta]-expressing peripheral T cells. 1990.

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Muthukumar, Thangamani, Darshana Dadhania, Choli Hartono und Manikkam Suthanthiran. Immunology, sensitization, and histocompatibility. Herausgegeben von Jeremy R. Chapman. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199592548.003.0279.

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Allograft rejection of the histo-incompatible allograft involves a highly orchestrated action of multiple cell types and mediators, with lymphocytes responsible for the identification of the foreignness of the allograft. The immune response directed against the donor is primarily, but not exclusively, directed at the donor’s major histocompatibility complex region class I and class II proteins. This chapter describes the immunobiology of the T cell and the role of human leucocyte antigens in clinical transplantation, thus identifying the targets for manipulation of the immune response by immune suppressants and through strategies designed to create a state of tolerance of the allograft.

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Liu, Stanley Kai-Wei. Identification and characterization of Gads - a novel hematopoietic-specific adaptor protein - in T cell receptor signaling. 2000.

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Rodríguez, Richard T. A Kiss across the Ocean. Duke University Press, 2022. http://dx.doi.org/10.1215/9781478023180.

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In A Kiss across the Ocean Richard T. Rodríguez examines the relationship between British post-punk musicians and their Latinx audiences in the United States since the 1980s. Melding memoir with cultural criticism, Rodríguez spotlights a host of influential bands and performers including Siouxsie and the Banshees, Adam Ant, Bauhaus, Soft Cell, Frankie Goes to Hollywood, and Pet Shop Boys. He recounts these bands’ importance for him and other Latinx kids and discusses their frequent identification with these bands’ glamorous performance of difference. Whether it was Siouxsie Sioux drawing inspiration from Latinx contemporaries and cultural practices or how Soft Cell singer Marc Almond’s lyrics were attuned to the vibrancy of queer Latinidad, Rodríguez shows how Latinx culture helped shape British post-punk. He traces the fandom networks that link these groups across space and time to illuminate how popular music establishes and facilitates intimate relations across the Atlantic. In so doing, he demonstrates how the music and styles that have come to define the 1980s hold significant sway over younger generations equally enthused by their matchlessly pleasurable and political reverberations.

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E, Menitove Jay, Kolins Jerry, American Association of Blood Banks. Committee on Technical/Scientific Workshops. und AIDS Technical Workshop (1986 : San Francisco, Calif.), Hrsg. AIDS. Arlington, Va: American Association of Blood Banks, 1986.

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Buchteile zum Thema "T cells Identification"

Gielis, Sofie, Pieter Moris, Wout Bittremieux, Nicolas De Neuter, Benson Ogunjimi, Kris Laukens und Pieter Meysman. „Identification of Epitope-Specific T Cells in T-Cell Receptor Repertoires“. In Bioinformatics for Cancer Immunotherapy, 183–95. New York, NY: Springer US, 2020. http://dx.doi.org/10.1007/978-1-0716-0327-7_13.

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Wacleche, Vanessa S., Runci Wang und Deepak A. Rao. „Identification of T Peripheral Helper (Tph) Cells“. In Methods in Molecular Biology, 59–76. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1736-6_6.

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Courey-Ghaouzi, Alan, und Mauro Gaya. „Identification of Non-classical Follicular T Cells“. In Methods in Molecular Biology, 77–84. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1736-6_7.

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Leehan, Kerry M., und Kristi A. Koelsch. „T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T Cells“. In Methods in Molecular Biology, 427–34. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2694-7_43.

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Beverley, P. C. L., M. Merkenschlager und D. L. Wallace. „Identification of Human Naive and Memory T Cells“. In Progress in Immunology, 432–38. Berlin, Heidelberg: Springer Berlin Heidelberg, 1989. http://dx.doi.org/10.1007/978-3-642-83755-5_57.

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Georgiev, Hristo, Georgia Papadogianni und Günter Bernhardt. „Identification of Follicular T Cells in the Gut“. In Methods in Molecular Biology, 85–95. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1736-6_8.

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Ding, Yanna, John D. Mountz und Hui-Chen Hsu. „Identification of Follicular T Helper Cells in Tissue Sections“. In Methods in Molecular Biology, 13–25. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2498-1_2.

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Koffeman, Eva, Elissa Keogh, Mark Klein, Berent Prakken und Salvatore Albani. „Identification and Manipulation of Antigen Specific T-Cells with Artificial Antigen Presenting Cells“. In Arthritis Research, 69–86. Totowa, NJ: Humana Press, 2007. http://dx.doi.org/10.1007/978-1-59745-402-5_6.

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Pedroso, Rodrigo, Filipa Ribeiro, Ana Rita Pires, Luis Graca und Valter R. Fonseca. „Identification of Human T Follicular Cells in Ectopic Lymphoid Structures“. In Methods in Molecular Biology, 225–33. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1736-6_19.

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Meli, Alexandre P., und Irah L. King. „Identification of Mouse T Follicular Helper Cells by Flow Cytometry“. In Methods in Molecular Biology, 3–11. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2498-1_1.

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Konferenzberichte zum Thema "T cells Identification"

Roos-Engstrand, Ester, Anders Bucht, Anders Blomberg, Annelie Behndig und Jamshid Pourazar. „Identification Of Regulatory T Cells In Stable COPD“. In American Thoracic Society 2010 International Conference, May 14-19, 2010 • New Orleans. American Thoracic Society, 2010. http://dx.doi.org/10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a3880.

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Naradikian, Martin S., Leslie Montero, Samantha Hall, Milad Bahmanof, Rukman Thota, Luise Sternberg, Jerome Lane et al. „Abstract 1439: Identification and characterization of neoantigen specific T cells“. In Proceedings: AACR Annual Meeting 2019; March 29-April 3, 2019; Atlanta, GA. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/1538-7445.sabcs18-1439.

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Codd, A., S. Al-Taei, S. Tokita, E. Mizushima, P. Rizkallah, T. Kanaseki, T. Torigoe, S. Man und Z. Tabi. „PO-407 Identification of unique antigens on prostate cancer stem cells for cytotoxic T cell recognition“. In Abstracts of the 25th Biennial Congress of the European Association for Cancer Research, Amsterdam, The Netherlands, 30 June – 3 July 2018. BMJ Publishing Group Ltd, 2018. http://dx.doi.org/10.1136/esmoopen-2018-eacr25.918.

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Mathieu, Mélissa, Sandy Pelletier, David Laperrière, Sylvie Mader und Simon Turcotte. „Abstract A084: Identification of mutation-reactive T cells in patients with gastrointestinal cancers“. In Abstracts: CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/2326-6074.cricimteatiaacr15-a084.

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Chen, Liang, Chunlin Wang und Mark Davis. „Abstract PR14: Identification of specificity TCR groups of tumor antigen specific T-cells“. In Abstracts: Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; September 30 - October 3, 2018; New York, NY. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/2326-6074.cricimteatiaacr18-pr14.

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Hudson, Will H., Julia L. Gensheimer, Haydn T. Kissick und Rafi Ahmed. „Abstract A196: Systematic identification of markers and drug targets on exhausted CD8+ T-cells“. In Abstracts: Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; September 30 - October 3, 2018; New York, NY. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/2326-6074.cricimteatiaacr18-a196.

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Chen, Jia-Shing, und H. Sunny Sun. „Abstract 3852: Identification and functional characterization of T-cell lymphoma invasion and metastasis 2 (TIAM2) in neuronal cells“. In Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.am2011-3852.

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Zong, Yunhui, Fuliang Chu, Songbing He, Yu Jing, Sally A. Hunsucker, Tina Yang, Ming Zhang et al. „Abstract 4094: Identification of co-inhibitory receptor expression on T cells from gastric cancer patients“. In Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA. American Association for Cancer Research, 2014. http://dx.doi.org/10.1158/1538-7445.am2014-4094.

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Bessede, Alban, Florent Peyraud, Jean Philippe Guegan, Christophe Rey, Sylvestre Le Moulec, Fabrice Barlesi, Sophie Cousin et al. „135 Identification of super-exhausted T cells: a novel population predictive of response to immunotherapy“. In SITC 37th Annual Meeting (SITC 2022) Abstracts. BMJ Publishing Group Ltd, 2022. http://dx.doi.org/10.1136/jitc-2022-sitc2022.0135.

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Berichte der Organisationen zum Thema "T cells Identification"

Baszler, Timothy, Igor Savitsky, Christopher Davies, Lauren Staska und Varda Shkap. Identification of bovine Neospora caninum cytotoxic T-lymphocyte epitopes for development of peptide-based vaccine. United States Department of Agriculture, März 2006. http://dx.doi.org/10.32747/2006.7695592.bard.

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The goal of the one-year feasibility study was to identify specific cytotoxic T-lymphocyte (CTL) epitopes to Neosporacaninum in the natural bovine host in order to make progress toward developing an effective peptide-based vaccine against bovine neosporosis. We tested the hypothesis that: N. caninum SRS2 peptides contain immunogenicCTLepitope clusters cross-presented by multiple bovine MHC-I and MHC-IIhaplotypes. The specific objectives were: (1) Map bovine CTLepitopes of N. caninum NcSRS-2 and identify consensus MHC-I and class-II binding motifs; and (2) Determine if subunit immunization with peptides containing N. caninum-specificCTLepitopes cross-reactive to multiple bovine MHChaplotypes induces a CTL response in cattle with disparate MHChaplotypes. Neosporosis is a major cause of infectious abortion and congenital disease in cattle, persisting in cattle herds via vertical transmission.5 N. caninum abortions are reported in Israel; a serological survey of 52 Israeli dairy herds with reported abortions indicated a 31% infection rate in cows and 16% infection rate in aborted fetuses.9,14 Broad economic loss due to bovine neosporosis is estimated at $35,000,000 per year in California, USA, and $100,000,000 (Australian) per year in Australia and New Zealand.13 Per herd losses in a Canadian herd of 50 cattle are estimated more conservatively at $2,305 (Canadian) annually.4 Up to date practical measures to reduce losses from neosporosis in cattle have not been achieved. There is no chemotherapy available and, although progress has been made toward understanding immunity to Neospora infections, no efficacious vaccine is available to limit outbreaks or prevent abortions. Vaccine development to prevent N. caninum abortion and congenital infection remains a high research priority. To this end, our research group has over the past decade: 1) Identified the importance of T-lymphocyte-mediated immunity, particularly IFN-γ responses, as necessary for immune protection to congenital neosporosis in mice,1,2,10,11 and 2) Identified MHC class II restricted CD4+ CTL in Neosporainfected Holstein cattle,16 and 3) Identified NcSRS2 as a highly conserved surface protein associated with immunity to Neospora infections in mice and cattle.7,8,15 In this BARD-funded 12 month feasibility study, we continued our study of Neospora immunity in cattle and successfully completed T-lymphocyte epitope mapping of NcSRS2 surface protein with peptides and bovine immune cells,15 fulfilling objective 1. We also documented the importance of immune responses NcSRS2 by showing that immunization with native NcSRS2 reduces congenital Neospora transmission in mice,7 and that antibodies to NcSRS2 specifically inhibition invasion of placental trophoblasts.8 Most importantly we showed that T-lymphocyte responses similar to parasite infection, namely induction of activated IFN-γ secreting Tlymphocytes, could be induced by subunit immunization with NcSRS2 peptides containing the Neospora-specificCTLepitopes (Baszler et al, In preparation) fulfilling objective 2. Both DNA and peptide-based subunit approaches were tested. Only lipopeptide-based NcSRS2 subunits, modified with N-terminal linked palmitic acid to enhance Toll-like receptors 2 and 1 (TLR2-TLR1), stimulated robust antigen-specific T-lymphocyte proliferation, IFN-γ secretion, and serum antibody production across different MHC-IIhaplotypes. The discovery of MHC-II cross-reactive T-cellinducing parasite peptides capable of inducing a potentially protective immune response following subunit immunization in cattle is of significant practical importance to vaccine development to bovine neosporosis. In addition, our findings are more widely applicable in future investigations of protective T-cell, subunit-based immunity against other infectious diseases in outbred cattle populations.

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Epel, Bernard, und Roger Beachy. Mechanisms of intra- and intercellular targeting and movement of tobacco mosaic virus. United States Department of Agriculture, November 2005. http://dx.doi.org/10.32747/2005.7695874.bard.

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To cause disease, plant viruses must replicate and spread locally and systemically within the host. Cell-to-cell virus spread is mediated by virus-encoded movement proteins (MPs), which modify the structure and function of plasmodesmata (Pd), trans-wall co-axial membranous tunnels that interconnect the cytoplasm of neighboring cells. Tobacco mosaic virus (TMV) employ a single MP for cell- cell spread and for which CP is not required. The PIs, Beachy (USA) and Epel (Israel) and co-workers, developed new tools and approaches for study of the mechanism of spread of TMV that lead to a partial identification and molecular characterization of the cellular machinery involved in the trafficking process. Original research objectives: Based on our data and those of others, we proposed a working model of plant viral spread. Our model stated that MPᵀᴹⱽ, an integral ER membrane protein with its C-terminus exposed to the cytoplasm (Reichel and Beachy, 1998), alters the Pd SEL, causes the Pd cytoplasmic annulus to dilate (Wolf et al., 1989), allowing ER to glide through Pd and that this gliding is cytoskeleton mediated. The model claimed that in absence of MP, the ER in Pd (the desmotubule) is stationary, i.e. does not move through the Pd. Based on this model we designed a series of experiments to test the following questions: -Does MP potentiate ER movement through the Pd? - In the presence of MP, is there communication between adjacent cells via ER lumen? -Does MP potentiate the movement of cytoskeletal elements cell to cell? -Is MP required for cell-to-cell movement of ER membranes between cells in sink tissue? -Is the binding in situ of MP to RNA specific to vRNA sequences or is it nonspecific as measured in vitro? And if specific: -What sequences of RNA are involved in binding to MP? And finally, what host proteins are associated with MP during intracellular targeting to various subcellular targets and what if any post-translational modifications occur to MP, other than phosphorylation (Kawakami et al., 1999)? Major conclusions, solutions and achievements. A new quantitative tool was developed to measure the "coefficient of conductivity" of Pd to cytoplasmic soluble proteins. Employing this tool, we measured changes in Pd conductivity in epidermal cells of sink and source leaves of wild-type and transgenic Nicotiana benthamiana (N. benthamiana) plants expressing MPᵀᴹⱽ incubated both in dark and light and at 16 and 25 ᵒC (Liarzi and Epel, 2005 (appendix 1). To test our model we measured the effect of the presence of MP on cell-to-cell spread of a cytoplasmic fluorescent probe, of two ER intrinsic membrane protein-probes and two ER lumen protein-probes fused to GFP. The effect of a mutant virus that is incapable of cell-to-cell spread on the spread of these probes was also determined. Our data shows that MP reduces SEL for cytoplasmic molecules, dilates the desmotubule allowing cell-cell diffusion of proteins via the desmotubule lumen and reduces the rate of spread of the ER membrane probes. Replicase was shown to enhance cell-cell spread. The data are not in support of the proposed model and have led us to propose a new model for virus cell-cell spread: this model proposes that MP, an integral ER membrane protein, forms a MP:vRNAER complex and that this ER-membrane complex diffuses in the lipid milieu of the ER into the desmotubule (the ER within the Pd), and spreads cell to cell by simple diffusion in the ER/desmotubule membrane; the driving force for spread is the chemical potential gradient between an infected cell and contingent non-infected neighbors. Our data also suggests that the virus replicase has a function in altering the Pd conductivity. Transgenic plant lines that express the MP gene of the Cg tobamovirus fused to YFP under the control the ecdysone receptor and methoxyfenocide ligand were generated by the Beachy group and the expression pattern and the timing and targeting patterns were determined. A vector expressing this MPs was also developed for use by the Epel lab . The transgenic lines are being used to identify and isolate host genes that are required for cell-to-cell movement of TMV/tobamoviruses. This line is now being grown and to be employed in proteomic studies which will commence November 2005. T-DNA insertion mutagenesis is being developed to identify and isolate host genes required for cell-to-cell movement of TMV.

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Lillehoj, Hyun, Dan Heller und Mark Jenkins. Cellular and molecular identification of Eimeria Acervulina Merozoite Antigens eliciting protective immunity. United States Department of Agriculture, November 1992. http://dx.doi.org/10.32747/1992.7561056.bard.

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Coccidiosis, ubiquitous diseases of poultry, seriously impair the growth and feed utilization of livestock and poultry. Coccidiosis causes over $600 million annual losses world-wide and no vaccine is currently available. The goal of this study was to investigate the cellular and molecular mechanisms controlling protective immune responses to coccidia parasites in order to develop immunological control strategy against coccidiosis. The major findings of this study were: 1) cell-mediated immunity plays a major role in protection against coccidiosis, 2) when different genetic lines showing different levels of disease susceptibility were compared, higher T-cell response was seen in the strains of chickens showing higher disease resistance, 3) early interferon secretion was observed in more coccidia-resistant chicken strains, 4) both sporozoite and merozoite antigens were able to induce interferon production, and 5) chicken monoclonal antibodies which detect immunogenic coccidia proteins have been developed. This study provided a good background work for future studies toward the development of recombinant coccidial vaccine. Availability of chicken monoclonal antibodies which detect immunogenic coccidia proteins will enhance our ability to identify potential coccidial vaccine antigens.

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Eldar, Avigdor, und Donald L. Evans. Streptococcus iniae Infections in Trout and Tilapia: Host-Pathogen Interactions, the Immune Response Toward the Pathogen and Vaccine Formulation. United States Department of Agriculture, Dezember 2000. http://dx.doi.org/10.32747/2000.7575286.bard.

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In Israel and in the U.S., Streptococcus iniae is responsible for considerable losses in various fish species. Poor understanding of its virulence factors and limited know-how-to of vaccine formulation and administration are the main reasons for the limited efficacy of vaccines. Our strategy was that in order to Improve control measures, both aspects should be equally addressed. Our proposal included the following objectives: (i) construction of host-pathogen interaction models; (ii) characterization of virulence factors and immunodominant antigens, with assessment of their relative importance in terms of protection and (iii) genetic identification of virulence factors and genes, with evaluation of the protective effect of recombinant proteins. We have shown that two different serotypes are involved. Their capsular polysaccharides (CPS) were characterized, and proved to play an important role in immune evasion and in other consequences of the infection. This is an innovative finding in fish bacteriology and resembles what, in other fields, has become apparent in the recent years: S. iniae alters surface antigens. By so doing, the pathogen escapes immune destruction. Immunological assays (agar-gel immunodiffusion and antibody titers) confirmed that only limited cross recognition between the two types occurs and that capsular polysaccharides are immunodominant. Vaccination with purified CPS (as an acellular vaccine) results in protection. In vitro and ex-vivo models have allowed us to unravel additional insights of the host-pathogen interactions. S. iniae 173 (type II) produced DNA fragmentation of TMB-8 cells characteristic of cellular necrosis; the same isolate also prevented the development of apoptosis in NCC. This was determined by finding reduced expression of phosphotidylserine (PS) on the outer membrane leaflet of NCC. NCC treated with this isolate had very high levels of cellular necrosis compared to all other isolates. This cellular pathology was confirmed by observing reduced DNA laddering in these same treated cells. Transmission EM also showed characteristic necrotic cellular changes in treated cells. To determine if the (in vitro) PCD/apoptosis protective effects of #173 correlated with any in vivo activity, tilapia were injected IV with #173 and #164 (an Israeli type I strain). Following injection, purified NCC were tested (in vitro) for cytotoxicity against HL-60 target cells. Four significant observations were made : (i) fish injected with #173 had 100-400% increased cytotoxicity compared to #164 (ii) in vivo activation occurred within 5 minutes of injection; (iii) activation occurred only within the peripheral blood compartment; and (iv) the isolate that protected NCC from apoptosis in vitro caused in vivo activation of cytotoxicity. The levels of in vivo cytotoxicity responses are associated with certain pathogens (pathogen associated molecular patterns/PAMP) and with the tissue of origin of NCC. NCC from different tissue (i.e. PBL, anterior kidney, spleen) exist in different states of differentiation. Random amplified polymorphic DNA (RAPD) analysis revealed the "adaptation" of the bacterium to the vaccinated environment, suggesting a "Darwinian-like" evolution of any bacterium. Due to the selective pressure which has occurred in the vaccinated environment, type II strains, able to evade the protective response elicited by the vaccine, have evolved from type I strains. The increased virulence through the appropriation of a novel antigenic composition conforms with pathogenic mechanisms described for other streptococci. Vaccine efficacy was improved: water-in-oil formulations were found effective in inducing protection that lasted for a period of (at least) 6 months. Protection was evaluated by functional tests - the protective effect, and immunological parameters - elicitation of T- and B-cells proliferation. Vaccinated fish were found to be resistant to the disease for (at least) six months; protection was accompanied by activation of the cellular and the humoral branches.

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Shpigel, Nahum, Raul Barletta, Ilan Rosenshine und Marcelo Chaffer. Identification and characterization of Mycobacterium paratuberculosis virulence genes expressed in vivo by negative selection. United States Department of Agriculture, Januar 2004. http://dx.doi.org/10.32747/2004.7696510.bard.

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Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of a severe inflammatory bowel disease (IBD) in ruminants, known as Johne’s disease or paratuberculosis. Johne’s disease is considered to be one of the most serious diseases affecting dairy cattle both in Israel and worldwide. Heavy economic losses are incurred by dairy farmers due to the severe effect of subclinical infection on milk production, fertility, lower disease resistance and early culling. Its influence in the United States alone is staggering, causing an estimated loss of $1.5 billion to the agriculture industry every year. Isolation of MAP from intestinal tissue and blood of Crohn's patients has lead to concern that it plays a potential pathogenic role in promoting human IDB including Crohn’s disease. There is great concern following the identification of the organism in animal products and shedding of the organism to the environment by subclinically infected animals. Little is known about the molecular basis for MAP virulence. The goal of the original proposed research was to identify MAP genes that are required for the critical stage of initial infection and colonization of ruminants’ intestine by MAP. We proposed to develop and use signature tag mutagenesis (STM) screen to find MAP genes that are specifically required for survival in ruminants upon experimental infection. This research projected was approved as one-year feasibility study to prove the ability of the research team to establish the animal model for mutant screening and alternative in-vitro cell systems. In Israel, neonatal goat kids were repeatedly inoculated with either one of the following organisms; MAP K-10 strain and three transposon mutants of K-10 which were produced and screened by the US PI. Six months after the commencement of inoculation we have necropsied the goats and taken multiple tissue samples from the jejunum, ileum and mesenteric lymph nodes. Both PCR and histopathology analysis indicated on efficient MAP colonization of all the inoculated animals. We have established several systems in the Israeli PI’s laboratory; these include using IS900 PCR for the identification of MAP and using HSP65-based PCR for the differentiation between MAV and MAP. We used Southern blot analysis for the differentiation among transposon mutants of K-10. In addition the Israeli PI has set up a panel of in-vitro screening systems for MAP mutants. These include assays to test adhesion, phagocytosis and survival of MAP to/within macrophages, assays that determine the rate of MAPinduced apoptosis of macrophages and MAP-induced NO production by macrophages, and assays testing the interference with T cell ã Interferon production and T cell proliferation by MAP infected macrophages (macrophage studies were done in BoMac and RAW cell lines, mouse peritoneal macrophages and bovine peripheral blood monocytes derived macrophages, respectively). All partners involved in this project feel that we are currently on track with this novel, highly challenging and ambitious research project. We have managed to establish the above described research systems that will clearly enable us to achieve the original proposed scientific objectives. We have proven ourselves as excellent collaborative groups with very high levels of complementary expertise. The Israeli groups were very fortunate to work with the US group and in a very short time period to master numerous techniques in the field of Mycobacterium research. The Israeli group has proven its ability to run this complicated animal model. This research, if continued, may elucidate new and basic aspects related to the pathogenesis MAP. In addition the work may identify new targets for vaccine and drug development. Considering the possibility that MAP might be a cause of human Crohn’s disease, better understanding of virulence mechanisms of this organism might also be of public health interest as well.

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Ron, Eliora, und Eugene Eugene Nester. Global functional genomics of plant cell transformation by agrobacterium. United States Department of Agriculture, März 2009. http://dx.doi.org/10.32747/2009.7695860.bard.

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The aim of this study was to carry out a global functional genomics analysis of plant cell transformation by Agrobacterium in order to define and characterize the physiology of Agrobacterium in the acidic environment of a wounded plant. We planed to study the proteome and transcriptome of Agrobacterium in response to a change in pH, from 7.2 to 5.5 and identify genes and circuits directly involved in this change. Bacteria-plant interactions involve a large number of global regulatory systems, which are essential for protection against new stressful conditions. The interaction of bacteria with their hosts has been previously studied by genetic-physiological methods. We wanted to make use of the new capabilities to study these interactions on a global scale, using transcription analysis (transcriptomics, microarrays) and proteomics (2D gel electrophoresis and mass spectrometry). The results provided extensive data on the functional genomics under conditions that partially mimic plant infection and – in addition - revealed some surprising and significant data. Thus, we identified the genes whose expression is modulated when Agrobacterium is grown under the acidic conditions found in the rhizosphere (pH 5.5), an essential environmental factor in Agrobacterium – plant interactions essential for induction of the virulence program by plant signal molecules. Among the 45 genes whose expression was significantly elevated, of special interest is the two-component chromosomally encoded system, ChvG/I which is involved in regulating acid inducible genes. A second exciting system under acid and ChvG/Icontrol is a secretion system for proteins, T6SS, encoded by 14 genes which appears to be important for Rhizobium leguminosarum nodule formation and nitrogen fixation and for virulence of Agrobacterium. The proteome analysis revealed that gamma aminobutyric acid (GABA), a metabolite secreted by wounded plants, induces the synthesis of an Agrobacterium lactonase which degrades the quorum sensing signal, N-acyl homoserine lactone (AHL), resulting in attenuation of virulence. In addition, through a transcriptomic analysis of Agrobacterium growing at the pH of the rhizosphere (pH=5.5), we demonstrated that salicylic acid (SA) a well-studied plant signal molecule important in plant defense, attenuates Agrobacterium virulence in two distinct ways - by down regulating the synthesis of the virulence (vir) genes required for the processing and transfer of the T-DNA and by inducing the same lactonase, which in turn degrades the AHL. Thus, GABA and SA with different molecular structures, induce the expression of these same genes. The identification of genes whose expression is modulated by conditions that mimic plant infection, as well as the identification of regulatory molecules that help control the early stages of infection, advance our understanding of this complex bacterial-plant interaction and has immediate potential applications to modify it. We expect that the data generated by our research will be used to develop novel strategies for the control of crown gall disease. Moreover, these results will also provide the basis for future biotechnological approaches that will use genetic manipulations to improve bacterial-plant interactions, leading to more efficient DNA transfer to recalcitrant plants and robust symbiosis. These advances will, in turn, contribute to plant protection by introducing genes for resistance against other bacteria, pests and environmental stress.

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Firon, Nurit, Prem Chourey, Etan Pressman, Allen Hartwell und Kenneth J. Boote. Molecular Identification and Characterization of Heat-Stress-Responsive Microgametogenesis Genes in Tomato and Sorghum - A Feasibility Study. United States Department of Agriculture, Oktober 2007. http://dx.doi.org/10.32747/2007.7591741.bard.

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Exposure to higher than optimal temperatures - heat-stress (HS) - is becoming increasingly common to all crop plants worldwide. Heat stress coinciding with microgametogenesis, especially during the post-meiotic phase that is marked by starch biosynthesis, is often associated with starch-deficient pollen and male sterility and ultimately, greatly reduced crop yields. The molecular basis for the high sensitivity of developing pollen grains, on one hand, and factors involved in pollen heat-tolerance, on the other, is poorly understood. The long-term goal of this project is to provide a better understanding of the genes that control pollen quality under heat-stress conditions. The specific objectives of this project were: (1) Determination of the threshold heat stress temperature(s) that affects tomato and sorghum pollen quality whether: a) Chronic mild heat stress conditions (CMHS), or b) Acute heat stress (AHS). (2) Isolation of heat-responsive, microgametogenesis-specific sequences. During our one-year feasibility project, we have accomplished the proposed objectives as follows: Objectrive 1: We have determined the threshold HS conditions in tomato and sorghum. This was essential for achieving the 2nd objective, since our accumulated experience (both Israeli and US labs) indicate that when temperature is raised too high above "threshold HS levels" it may cause massive death of the developing pollen grains. Above-threshold conditions have additional major disadvantages including the "noise" caused by induced expression of genes involved in cell death and masking of the differences between heatsensitive and heat-tolerant pollen grains. Two different types of HS conditions were determined: a) Season-long CMHS conditions: 32/26°C day/night temperatures confirmed in tomato and 36/26°C day maximum/night minimum temperatures in sorghum. b) Short-term AHS: In tomato, 2 hour exposure to 42-45°C (at 7 to 3 days before anthesis) followed by transfer to 28/22±2oC day/night temperatures until flower opening and pollen maturation, caused 50% reduced germinating pollen in the heat-sensitive 3017 cv.. In sorghum, 36/26°C day/night temperatures 10 to 5 days prior to panicle emergence, occurring at 35 days after sowing (DAS) in cv. DeKalb28E, produced starch-deficient and sterile pollen. Objective 2: We have established protocols for the high throughput transcriptomic approach, cDNA-AFLP, for identifying and isolating genes exhibiting differential expression in developing microspores exposed to either ambient or HS conditions and created a databank of HS-responsivemicrogametogenesis-expressed genes. A subset of differentially displayed Transcript-Derived Fragments (TDFs) that were cloned and sequenced (35 & 23 TDFs in tomato and sorghum, respectively) show close sequence similarities with metabolic genes, genes involved in regulation of carbohydrate metabolism, genes implicated in thermotolerance (heat shock proteins), genes involved in long chain fatty acids elongation, genes involved in proteolysis, in oxidation-reduction, vesicle-mediated transport, cell division and transcription factors. T-DNA-tagged Arabidopsis mutants for part of these genes were obtained to be used for their functional analysis. These studies are planned for a continuation project. Following functional analyses of these genes under HS – a valuable resource of genes, engaged in the HS-response of developing pollen grains, that could be modulated for the improvement of pollen quality under HS in both dicots and monocots and/or used to look for natural variability of such genes for selecting heat-tolerant germplasm - is expected.

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Altstein, Miriam, und Ronald J. Nachman. Rational Design of Insect Control Agent Prototypes Based on Pyrokinin/PBAN Neuropeptide Antagonists. United States Department of Agriculture, August 2013. http://dx.doi.org/10.32747/2013.7593398.bard.

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The general objective of this study was to develop rationally designed mimetic antagonists (and agonists) of the PK/PBAN Np class with enhanced bio-stability and bioavailability as prototypes for effective and environmentally friendly pest insect management agents. The PK/PBAN family is a multifunctional group of Nps that mediates key functions in insects (sex pheromone biosynthesis, cuticular melanization, myotropic activity, diapause and pupal development) and is, therefore, of high scientific and applied interest. The objectives of the current study were: (i) to identify an antagonist biophores (ii) to develop an arsenal of amphiphilic topically active PK/PBAN antagonists with an array of different time-release profiles based on the previously developed prototype analog; (iii) to develop rationally designed non-peptide SMLs based on the antagonist biophore determined in (i) and evaluate them in cloned receptor microplate binding assays and by pheromonotropic, melanotropic and pupariation in vivo assays. (iv) to clone PK/PBAN receptors (PK/PBAN-Rs) for further understanding of receptor-ligand interactions; (v) to develop microplate binding assays for screening the above SMLs. In the course of the granting period A series of amphiphilic PK/PBAN analogs based on a linear lead antagonist from the previous BARD grant was synthesized that incorporated a diverse array of hydrophobic groups (HR-Suc-A[dF]PRLa). Others were synthesized via the attachment of polyethylene glycol (PEG) polymers. A hydrophobic, biostablePK/PBAN/DH analog DH-2Abf-K prevented the onset of the protective state of diapause in H. zea pupae [EC50=7 pmol/larva] following injection into the preceding larval stage. It effectively induces the crop pest to commit a form of ‘ecological suicide’. Evaluation of a set of amphiphilic PK analogs with a diverse array of hydrophobic groups of the formula HR-Suc-FTPRLa led to the identification of analog T-63 (HR=Decyl) that increased the extent of diapause termination by a factor of 70% when applied topically to newly emerged pupae. Another biostablePK analog PK-Oic-1 featured anti-feedant and aphicidal properties that matched the potency of some commercial aphicides. Native PK showed no significant activity. The aphicidal effects were blocked by a new PEGylated PK antagonist analog PK-dF-PEG4, suggesting that the activity is mediated by a PK/PBAN receptor and therefore indicative of a novel and selective mode-of-action. Using a novel transPro mimetic motif (dihydroimidazole; ‘Jones’) developed in previous BARD-sponsored work, the first antagonist for the diapause hormone (DH), DH-Jo, was developed and shown to block over 50% of H. zea pupal diapause termination activity of native DH. This novel antagonist development strategy may be applicable to other invertebrate and vertebrate hormones that feature a transPro in the active core. The research identifies a critical component of the antagonist biophore for this PK/PBAN receptor subtype, i.e. a trans-oriented Pro. Additional work led to the molecular cloning and functional characterization of the DH receptor from H. zea, allowing for the discovery of three other DH antagonist analogs: Drosophila ETH, a β-AA analog, and a dF analog. The receptor experiments identified an agonist (DH-2Abf-dA) with a maximal response greater than native DH. ‘Deconvolution’ of a rationally-designed nonpeptide heterocyclic combinatorial library with a cyclic bis-guanidino (BG) scaffold led to discovery of several members that elicited activity in a pupariation acceleration assay, and one that also showed activity in an H. zea diapause termination assay, eliciting a maximal response of 90%. Molecular cloning and functional characterization of a CAP2b antidiuretic receptor from the kissing bug (R. prolixus) as well as the first CAP2b and PK receptors from a tick was also achieved. Notably, the PK/PBAN-like receptor from the cattle fever tick is unique among known PK/PBAN and CAP2b receptors in that it can interact with both ligand types, providing further evidence for an evolutionary relationship between these two NP families. In the course of the granting period we also managed to clone the PK/PBAN-R of H. peltigera, to express it and the S. littoralis-R Sf-9 cells and to evaluate their interaction with a variety of PK/PBAN ligands. In addition, three functional microplate assays in a HTS format have been developed: a cell-membrane competitive ligand binding assay; a Ca flux assay and a whole cell cAMP ELISA. The Ca flux assay has been used for receptor characterization due to its extremely high sensitivity. Computer homology studies were carried out to predict both receptor’s SAR and based on this analysis 8 mutants have been generated. The bioavailability of small linear antagonistic peptides has been evaluated and was found to be highly effective as sex pheromone biosynthesis inhibitors. The activity of 11 new amphiphilic analogs has also been evaluated. Unfortunately, due to a problem with the Heliothis moth colony we were unable to select those with pheromonotropic antagonistic activity and further check their bioavailability. Six peptides exhibited some melanotropic antagonistic activity but due to the low inhibitory effect the peptides were not further tested for bioavailability in S. littoralis larvae. Despite the fact that no new antagonistic peptides were discovered in the course of this granting period the results contribute to a better understanding of the interaction of the PK/PBAN family of Nps with their receptors, provided several HT assays for screening of libraries of various origin for presence of PK/PBAN-Ragonists and antagonists and provided important practical information for the further design of new, peptide-based insecticide prototypes aimed at the disruption of key neuroendocrine physiological functions in pest insects.

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Lers, Amnon, und Pamela J. Green. Analysis of Small RNAs Associated with Plant Senescence. United States Department of Agriculture, März 2013. http://dx.doi.org/10.32747/2013.7593393.bard.

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Senescence is an agriculturally significant process due to its negative impact to crop yield and postharvest quality. The genetic regulatory systems controlling senescence induction and progress respond to both developmental and environmental stress signals and involve numerous gene expression changes. Knowledge about the key molecular factors which control senescence is very limited. MicroRNAs (miRNAs) are a class of small RNAs which typically function by guiding cleavage of target messenger RNAs. They have been shown to play major roles in a variety of plant processes including development, responses to environmental stresses, and senescence. The long-term goal of this work is to elucidate roles of small RNAs associated with plant senescence. The hypothesis underlying this research is that miRNA-mediated regulation makes important contributions to the senescence process in plants. Specific, original research objectives included: 1) Profiling of small RNAs from senescing plants; 2) Data Analysis and public access via a user-friendly web interface; 3) Validation of senescence-associated miRNAs and target RNAs; 4) Development of transgenic plants for functional analysis of miRNAs in Arabidopsis. Major revisions made in the research compared to the original work plan included 1) Exclusion of the planned work with tomato as recommended by the BARD review panel; 2) Performing miRNA study also in senescing Arabidopsis siliques, in addition to senescing leaves. To identify senescenceregulation of miRNAs in Arabidopsis thaliana, eight small RNA libraries were constructed and sequenced at four different stages of development and senescence from both leaves and siliques, resulting in more than 200 million genome-matched sequences. Parallel Analysis of RNA Ends (PARE) libraries, which enable the large-scale examination of miRNA-guided cleavage products, were also constructed and sequenced, resulting in over 750 million genome-matched sequences. These massive datasets lead to the identification of new miRNAs, as well as new regulation of known miRNAs and their target genes during senescence, many of which have established roles in nutrient responsiveness and cell structural integrity. In keeping with remobilization of nutrients thought to occur during senescence, many miRNAs and targets had opposite expression pattern changes between leaf and silique tissues during the progression of senescence. Taken together, these findings highlight the integral role that miRNAs may play in the remobilization of resources and alteration of cellular structure that is known to occur in senescence. Experiments were initiated for functional analysis of specific senescence-associated miRNAs and respective target genes. Transgenic Arabidopsis plants were generated in which miR408, found in this study to be significantly induced in leaf senescence, was over-expressed either constitutively or under a senescence-specific promoter. These plants are currently being characterized for any altered phenotypes. In addition T-DNA knock out mutants for various target genes identified in this research are being analyzed. This work provides insights about specific miRNAs that contribute to leaf and silique senescence. The knowledge generated may suggest new strategies to monitor and alter the progression of senescence in crops for agricultural improvement.

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