Bibliografías temáticas / Antibody Secreting Cell

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Literatura académica sobre el tema "Antibody Secreting Cell"

Autor: Grafiati

Publicado: 10 de mayo de 2025

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Artículos de revistas sobre el tema "Antibody Secreting Cell"

Lin, Ling, Andrea J. Gerth y Stanford L. Peng. "Active Inhibition of Plasma Cell Development in Resting B Cells by Microphthalmia-associated Transcription Factor". Journal of Experimental Medicine 200, n.º 1 (28 de junio de 2004): 115–22. http://dx.doi.org/10.1084/jem.20040612.

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B cell terminal differentiation involves development into an antibody-secreting plasma cell, reflecting the concerted activation of proplasma cell transcriptional regulators, such as Blimp-1, IRF-4, and Xbp-1. Here, we show that the microphthalmia-associated transcription factor (Mitf) is highly expressed in naive B cells, where it antagonizes the process of terminal differentiation through the repression of IRF-4. Defective Mitf activity results in spontaneous B cell activation, antibody secretion, and autoantibody production. Conversely, ectopic Mitf expression suppresses the expression of IRF-4, the plasma cell marker CD138, and antibody secretion. Thus, Mitf regulates B cell homeostasis by suppressing the antibody-secreting fate.

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Yu, Tian, Jonathan Hull, Andrea Ruiz, Ashwini Bhat y Amar Basu. "Expediting antibody discovery using Bioelectronica’s HypercellTM platform". Journal of Immunology 204, n.º 1_Supplement (1 de mayo de 2020): 86.36. http://dx.doi.org/10.4049/jimmunol.204.supp.86.36.

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Abstract Antibody-based drugs have been successful in a range of therapeutic categories. However, generating monoclonal antibodies is time-consuming and expensive. A common approach is Hybridoma technology, which overcomes the short life-span of IgG-secreting plasma B cells in vitro. However, many plasma B cells are lost due to the low efficiency of hybridoma cell fusion (typically <10%). Direct single B cell screening strategies have emerged to bypass hybridoma fusion and recombinatorial display, coupled with the generation of recombinant monoclonal antibodies through mammalian expression systems. Obtaining expression systems with the required productivity, specificity and stability for clinical or commercial use requires screening millions of cells and thousands of clones. Bioelectronica’s HypercellTM platform is an emerging technology used throughout antibody discovery and cell-line development to identify and isolate single, high-antibody secreting cells from large pools (~10,000,000 cells) in short periods (ca. 48 hrs). This scalable “electrofluidic” sorting system reduces time and cost by integrating antigen-detection reagents and real-time computer vision analysis to expedite single cell sorting. In this paper antigen-specific IgG-secreting hybridoma cells are identified and sorted by their secretion rate. The cells and reagents are encapsulated in a Polydisperse Oblate Dispersion system (PODs), incubated for 1–4 hours for signal gain, and loaded into the HypercellTM device for cell sorting. Alternatively, the mixture can be analyzed without sorting to produce single cell secretion “finger print” signatures that can help identify unique expression patterns and monitor cell line stability over culturing time.

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Andreani, Virginia, Senthilkumar Ramamoorthy, Abhinav Pandey, Ekaterina Lupar, Stephen L. Nutt, Tim Lämmermann y Rudolf Grosschedl. "Cochaperone Mzb1 is a key effector of Blimp1 in plasma cell differentiation and β1-integrin function". Proceedings of the National Academy of Sciences 115, n.º 41 (26 de septiembre de 2018): E9630—E9639. http://dx.doi.org/10.1073/pnas.1809739115.

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Plasma cell differentiation involves coordinated changes in gene expression and functional properties of B cells. Here, we study the role of Mzb1, a Grp94 cochaperone that is expressed in marginal zone (MZ) B cells and during the terminal differentiation of B cells to antibody-secreting cells. By analyzing Mzb1−/−Prdm1+/gfp mice, we find that Mzb1 is specifically required for the differentiation and function of antibody-secreting cells in a T cell-independent immune response. We find that Mzb1-deficiency mimics, in part, the phenotype of Blimp1 deficiency, including the impaired secretion of IgM and the deregulation of Blimp1 target genes. In addition, we find that Mzb1−/− plasmablasts show a reduced activation of β1-integrin, which contributes to the impaired plasmablast differentiation and migration of antibody-secreting cells to the bone marrow. Thus, Mzb1 function is required for multiple aspects of plasma cell differentiation.

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Golding, B., S. R. Pillemer, P. Roussou, E. A. Peters, G. C. Tsokos, J. E. Ballow y T. Hoffman. "Inverse relationship between proliferation and differentiation in a human TNP-specific B cell line. Cell cycle dependence of antibody secretion." Journal of Immunology 141, n.º 8 (15 de octubre de 1988): 2564–68. http://dx.doi.org/10.4049/jimmunol.141.8.2564.

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Abstract Studies of an EBV-transformed and TNP-specific human B cell line revealed that, unlike myeloma or hybridoma cell lines that consist mainly of fully differentiated cells, most of the cloned EBV-transformed cells were not fully differentiated, as judged by inability to bind TNP-SRBC and to secrete anti-TNP antibody. The minority of more differentiated cells were selected by TNP-SRBC rosetting. They were found to proliferate to a lesser extent than nonrosetting cells and to contain increased numbers of antibody-secreting cells. This inverse relationship between proliferation and differentiation was also shown to be cell cycle related in that the TNP-SRBC rosetting cells resided, to a greater extent than the nonrosetting cells, in the G1 phase of the cell cycle. The finding that the G1 phase of the cell cycle was associated with differentiation into anti-TNP secreting cells was confirmed by demonstrating that treatment with hydroxyurea, which arrests the cells in G1, resulted in decreased proliferation and an increased proportion of antibody-secreting cells. Similarly, addition of phorbol ester resulted in increased antibody secretion and decreased proliferation, suggesting a role for protein kinase C in this differentiation pathway. The strategy of increasing the number of antibody-producing cells in this human EBV line, by promoting differentiation of the cells in G1, may be relevant to the large scale production of specific human mAb for the treatment and diagnosis of human diseases.

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Keane, John T. y Avery D. Posey. "Abstract 4089: PanCAR-specific antibody-cytokine fusion proteins". Cancer Research 83, n.º 7_Supplement (4 de abril de 2023): 4089. http://dx.doi.org/10.1158/1538-7445.am2023-4089.

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Abstract CAR-T cells are a potent immunotherapy that redirect T cell specificity towards cell surface tumor-associated antigens. While this therapy has been effective in hematological malignancies, therapies against solid tumors not achieved the same efficacy due to a multitude of additional challenges, including a lack of tumor specific antigens and an immunosuppressive tumor microenvironment. One strategy to increase T cell potency is to use fourth generation CAR-T cells that also secrete an immunostimulatory factor, such as a cytokine. We have created CAR-T cells targeting Tn-MUC1 utilizing a scFv from the 5E5 antibody that also secrete interleukin-12 (IL-12), interleukin-18 (IL-18), and interleukin-23 (IL-23). These cytokine-secreting CAR-T cells have shown an increase in efficacy against multiple epithelial based tumors in impedance-based killing assays. In a xenograft model of human breast cancer, CAR-T cells secreting IL-12 and IL-23 were able to significantly delay tumor growth compared to non-transduced T cells, CAR-T cells alone, and CAR-T cells secreting IL-18. While these cells showed good efficacy in vivo, clinical translation is difficult due to concerns over cytokine storm due to the constitutive secretion of cytokines. To overcome this, we created a fusion protein utilizing an antibody specific to an invariant region found in majority of CAR molecules, including those utilized commercially available therapies and under pre-clinical development, fused to the three immunostimulatory cytokines. In impedance-based killing assays, antibody-cytokine fusions stimulated non-cytokine secreting CAR-T cells to completely clear the tumor at an E:T ratio in which the CAR-T cells incubated with only the antibody alone were unable to control the tumor growth. Interestingly, combination therapy of CAR-T cells and antibody-cytokine fusions outperformed CAR-T cells constitutively secreting the same cytokines in in vitro assays. This work highlights the use of a single reagent capable of increasing the efficacy of the majority of CAR-T cell therapies and may be an important contribution towards treating solid tumor malignancies with CAR-T cell therapies. Citation Format: John T. Keane, Avery D. Posey. PanCAR-specific antibody-cytokine fusion proteins. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4089.

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Hoogenboom, H. R., J. C. Raus y G. Volckaert. "Cloning and expression of a chimeric antibody directed against the human transferrin receptor." Journal of Immunology 144, n.º 8 (15 de abril de 1990): 3211–17. http://dx.doi.org/10.4049/jimmunol.144.8.3211.

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Abstract The cloning, construction and expression of chimeric Ig genes, encoding a mAb directed against the human transferrin receptor, is described. From a mouse hybridoma cell line, secreting an antitransferrin receptor antibody, mRNA was prepared and converted into cDNA using Ig-specific oligonucleotides. H and L chain encoding cDNA fragments were isolated and sequenced. Chimeric genes were constructed by linking the murine V region cDNA fragments to human C region exons. After sequential transfection of nonproducing mouse hybridoma cells with the expression vectors containing the chimeric H and L chain genes, antibody secreting transfectomas were obtained. ELISA and immunoblot analysis clearly demonstrate the secretion of human kappa- and gamma-1 chain. Flow microfluorimetry analysis of the chimeric antibody shows that the Ag-binding capacity has been retained. The chimeric antibody most likely will be less immunogenic then the original mouse antibody when used in human cancer therapy.

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McHeyzer-Williams, M. G. y G. J. Nossal. "Clonal analysis of autoantibody-producing cell precursors in the preimmune B cell repertoire." Journal of Immunology 141, n.º 12 (15 de diciembre de 1988): 4118–23. http://dx.doi.org/10.4049/jimmunol.141.12.4118.

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Abstract Evidence for anti-self-reactivity in the preimmune B cell repertoire has been well documented. The present study aimed to determine the frequency of antibody-forming cell precursors in this repertoire whose Ig V regions impart reactivity to "self" or autologous Ag. Clones were activated in vitro with LPS and their secreted IgM antibody was assayed for reactivity by direct binding to cell surface or intracellular Ag. An IL-4-containing lymphokine mixture was added to the clonal cultures to induce the secretion of IgG1. The reactivity of secreted IgG1 with Ag would more closely resemble the binding required to activate B cells through their monomeric surface IgM and/or IgD. The results indicate a high frequency of precursors secreting IgM with reactivity to intracellular Ag, namely 1 in 37 +/- 6 B cells, with a marked paucity of response to cell surface molecules. The repertoire was markedly deficient in precursors secreting IgG1 able to bind to intracellular Ag, with only one clone detected by the screening of 3.0 x 10(6) spleen cells. No positives were detected for cell surface Ag. This suggested that the frequency of clones in the preimmune repertoire that express IgR with sufficient affinity to bind "self" molecules must be very low.

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Marquez, C., M. L. Toribio, M. A. Marcos, A. de la Hera, A. Barcena, L. Pezzi y C. Martinez. "Expression of alkaline phosphatase in murine B lymphocytes. Correlation with B cell differentiation into Ig secretion." Journal of Immunology 142, n.º 9 (1 de mayo de 1989): 3187–92. http://dx.doi.org/10.4049/jimmunol.142.9.3187.

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Abstract Alkaline phosphatases (ALPase) (orthophosphoric-monoester phosphohydrolase, EC 3.1.3.1) are implicated in many biologic phenomena including ossification and differentiation of human neutrophils and choriocarcinoma cells. Another trait, demonstrated by microinjection into Xenopus oocytes, is their ability to block the first mitotic division. Previous work in our laboratory has established that ALPase is also present on murine B lymphocytes activated by either polyclonal mitogens or Th cells. We have now characterized the ALPase present on murine B cells as belonging to the liver-bone-kidney isoenzyme and found it to be implicated in B cell differentiation into antibody secretion. Thus, B cell proliferative responses, elicited either by high concentrations of rabbit anti-IgM antibodies or by LPS in the presence of PMA, are characterized by the lack of both antibody secretion and expression of ALPase activity. In contrast, B cells stimulated to differentiate into Ig-secreting cells by B cell differentiation factors, nearly in the absence of a proliferative response, express high levels of ALPase activity, as did those that were LPS-stimulated. These data showing the association of the ALPase expression with the process of B cell differentiation into antibody-secreting cells are discussed in the context of the possible role that phosphorylation-dephosphorylation mechanism may play in controlling the growth/differentiation rate in the B cell lineage.

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Sankari, Hanna, Minna Hietikko, Kalle Kurppa, Katri Kaukinen, Eriika Mansikka, Heini Huhtala, Kaija Laurila et al. "Intestinal TG3- and TG2-Specific Plasma Cell Responses in Dermatitis Herpetiformis Patients Undergoing a Gluten Challenge". Nutrients 12, n.º 2 (13 de febrero de 2020): 467. http://dx.doi.org/10.3390/nu12020467.

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Dermatitis herpetiformis (DH), a cutaneous manifestation of coeliac disease, is characterized by transglutaminase (TG) 3-targeted dermal immunoglobulin A (IgA) deposits. The treatment for DH is the same as for coeliac disease, namely a life-long gluten-free diet. DH patients typically have gluten-dependent circulating autoantibodies targeting TG3 and TG2, and plasma cells secreting such autoantibodies have been detected in the small intestinal mucosa. This study investigates the gluten-responsiveness of intestinal TG3 and TG2 antibody-secreting plasma cells in 16 treated DH patients undergoing a gluten challenge. The frequency of both plasma cell populations increased significantly during the challenge, and their frequency correlated with the corresponding serum autoantibody levels at post-challenge. TG3-specific plasma cells were absent in all 18 untreated coeliac disease patients and seven non-coeliac control subjects on gluten-containing diets. These findings indicate that, in DH, both intestinal TG3- and TG2-antibody secreting plasma cells are gluten-dependent, and that TG3-antibody secreting plasma cells are DH-specific.

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Kirchenbaum, Greg A., Giuseppe A. Sautto, Rodrigo B. Abreu, Paul V. Lehmann y Ted M. Ross. "Assessment of Antibody Functional Affinity using FluoroSpot". Journal of Immunology 204, n.º 1_Supplement (1 de mayo de 2020): 86.11. http://dx.doi.org/10.4049/jimmunol.204.supp.86.11.

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Abstract Studies of B cell immunity often rely upon serologic methodologies that primarily assess antibody specificity. However, functional affinity of the antigen-specific antibody repertoire is a key determinant of protective efficacy. Presently, detailed assessment of single B cell/antibody functional affinity is labor-intensive and low-throughput. Therefore, we sought to evaluate whether B cell FluoroSpot assays would enable distinction between B cells with differential functional affinity since fluorescent intensity is directly proportional to antibody abundance in this assay. To test our prediction, murine B cell hybridomas (P65C6-2, 36–65 and 36–71) secreting monoclonal antibodies (mAb) with differential affinity for the p-azophenylarsonate (Ars) hapten were utilized as model antibody-secreting cells (ASC). Additionally, usage of an anti-idiotypic mAb (17–63) specific for the high-affinity anti-Ars mAb (36–71) confirmed unambiguous segregation of Ars-specific ASC. Moreover, we also evaluated the capacity of a multiplexed FluoroSpot assay to simultaneously distinguish antibody functional affinity among influenza hemagglutinin-reactive murine B cell hybridomas, or in vivo differentiated ASC from immunized mice, secreting various IgG subclasses (IgG1/IgG2a/IgG2b) in parallel. Collectively, our data support the notion that FluoroSpot assays can be used to assess the functional affinity of antigen-specific B cells, and that this approach is well-suited for detailed assessment of humoral immunity.

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Tesis sobre el tema "Antibody Secreting Cell"

Rousseau, Fanny. "Systèmes microfluidiques pour la génération d'hybridomes et d'anticorps monoclonaux". Electronic Thesis or Diss., université Paris-Saclay, 2025. http://www.theses.fr/2025UPASQ013.

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Les anticorps sont des molécules produites par le système immunitaire, caractérisées par une grande spécificité de liaison pour un antigène donné, ce qui en fait des outils biologiques puissants pour des applications thérapeutiques et de diagnostic. La production en quantité d'anticorps monoclonaux in vitro est rendue possible en 1975 grâce à la technique des hybridomes. Bien que cette technique soit simple, facile à implémenter et peu coûteuse, ses faibles rendements limitent aujourd'hui son utilisation et ont donné lieu à des méthodes plus modernes de production d'anticorps, qui présentent toutefois de nouvelles limitations.Dans ce contexte, l'objectif de cette thèse est d'améliorer la technique des hybridomes actuelle afin de la repositionner comme une technologie innovante et attractive. En particulier, il s'agit d'implémenter trois dispositifs microfluidiques, intervenant aux différentes étapes du processus de production d'anticorps monoclonaux, pour en optimiser les rendements et en faciliter la procédure.La première partie de ce projet est consacrée à l'identification et à l'isolement des cellules sécrétrices d'anticorps issues de rates de souris immunisées. Après fusion avec les cellules de myélome, ces cellules favorisent la génération d'hybridomes sécrétant des anticorps spécifiques de l'antigène cible. Le but est ainsi de pouvoir réaliser des fusions cellulaires ciblées pour limiter la génération d'hybridomes non fonctionnels. Pour cela, après identification par cytométrie en flux à l'aide d'un panel de marqueurs membranaires ad hoc, l'isolement des cellules d'intérêt est réalisé à l'aide d'un tri magnétique intégré en puce microfluidique.La deuxième partie de la thèse porte sur le développement d'une puce microfluidique dédiée à la fusion cellulaire par voie chimique, en utilisant le polyéthylène glycol (PEG). Ce dispositif vise à optimiser les conditions de fusion entre des cellules de rate et de myélome, et à améliorer les rendements de la méthode conventionnelle. Une version alternative de cette puce, adaptée à la fusion cellulaire par électroporation est également démontrée.Enfin, la dernière partie de cette thèse est consacrée à la sélection des hybridomes sécréteurs d'anticorps spécifiques de la cible d'intérêt par criblage en microfluidique de gouttes. Cette partie, réalisée en collaboration avec l'entreprise strasbourgeoise MicroOmix, permet de simplifier et d'accélérer les étapes post fusion de sélection des clones d'intérêt employées dans la technique des hybridomes
Antibodies are molecules produced by the immune system and are characterized by their high binding affinity and specificity for a given antigen, thus making them powerful biological tools for therapeutic and diagnostic applications. The in vitro production of antibodies was made possible in 1975 by the development of the hybridoma technology. This technique is simple, easy to implement and inexpensive, but its use has been limited by low yields, which has led to the emergence of more modern methods that present their own set of challenges.The central aim of this thesis is to unlock the existing hybridoma technique, repositioning it as an efficient and appealing technology. In particular, the objective is to implement three microfluidic devices at each step of the monoclonal antibody production process in order to optimise yields and facilitate the procedure.The first part of this project is focused on the identification and isolation of antibody-secreting cells from the spleen of immunized mice. Following fusion with myeloma cells, this cell subset may facilitate the generation of hybridomas with the ability to secret antibodies that are specific to the target antigen. Our aim is thus to perform targeted fusions between myeloma and antibody-secreting cells, thereby preventing the generation of non-functional hybridomas. To achieve this objective, the cells of interest are identified by flow cytometry using a dedicated surface markers panel. These cells subset are then isolated using an integrated microfluidic magnetic cell-sorting chip.The second part of the thesis concerns the development of a microfluidic chip dedicated to chemical cell fusion, using polyethylene glycol (PEG). The objective of this device is to optimize the conditions for efficient fusion between spleen and myeloma cells, and to improve the yields of the conventional method in tube. An alternative version of this chip, adapted for cell fusion by electroporation, is also demonstrated.Finally, the last part of this project illustrates the potential of droplet-based microfluidics for the single-cell selection and high-throughput screening of hybridomas that secret antigen-specific antibodies. This demonstration, carried out in collaboration with the Strasbourg-based company MicroOmix, aims to simplify and accelerate the post-fusion steps of the hybridoma technology

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Atkinson, Jeffrey Ross. "Peripheral Germinal Centers Regulate Virus-Specific B Cell Accumulation in the CNS". Kent State University / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=kent1524683244217474.

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VanCott, John Louis. "Protective immunity against transmissible gastroenteritis virus (TGEW) : enumeration of antibody-secreting cells and identification of mononuclear cell surface markers in systemic and mucosal lymphoid tissues of young pigs exposed to TGEV... /". The Ohio State University, 1993. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487842372895095.

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Herbert, Joan. "The regulation of specific antibody secretion by human B cells through contact and non-contact dependent mechanisms". Thesis, University College London (University of London), 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.244614.

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Law, Yuet Ching. "Specific compartmentalization of Immunoglobulin A antibody secreting cells in mouse salivary glands via the differential expression of chemokines and chemokine receptors /". Diss., CLICK HERE for online access, 2008. http://contentdm.lib.byu.edu/ETD/image/etd2678.pdf.

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Wennhold, Kerstin [Verfasser] y Thorsten [Akademischer Betreuer] Hoppe. "Tumorantigen-Specific CD40B Cells: Combining Enhanced Antigen-Presentation and Antibody-Secretion for Tumor Targeting / Kerstin Wennhold. Gutachter: Thorsten Hoppe". Köln : Universitäts- und Stadtbibliothek Köln, 2015. http://d-nb.info/1071369849/34.

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Nieminen, Tea. "Circulating antibody-secreting cells and salivary antibodies induced by the capsular polysaccharide of Streptococcus pneumoniae : after parenteral immunisation and in acute otitis media". Helsinki : University of Helsinki, 1999. http://ethesis.helsinki.fi/julkaisut/laa/haart/vk/nieminen/.

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Kiran, Kumar Mudnakudu Nagaraju [Verfasser], Thomas [Akademischer Betreuer] Friedrich, Margitta [Akademischer Betreuer] Worm y Roderich [Akademischer Betreuer] Süssmuth. "Targeting antibody secreting B cells as a novel therapeutic approach to treat allergic diseases / Mudnakudu Nagaraju Kiran Kumar. Gutachter: Margitta Worm ; Roderich Süssmuth. Betreuer: Thomas Friedrich". Berlin : Technische Universität Berlin, 2014. http://d-nb.info/1066162840/34.

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Strannermyr, Malin. "Increased expression of proteins in CHO cells by identification of signal peptides for improved secretion of translated proteins". Thesis, Linköpings universitet, Teknisk biologi, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-150364.

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Main purpose of this study was to increase protein expression in Chinese hamster ovary (CHO) cells by improving protein secretion of translated proteins. The goal was to find signal peptides from the screening of signal peptide libraries for improvement of protein secretion using a CHO-cell express selection system. Biopharmaceutical products, proteins such as monoclonal antibodies (mAbs), are most commonly produced using mammalian expression systems such as the expression in CHO cells. The posttranslational modifications of the proteins being expressed in CHO cells are similar to the expressional modifications in human cells, why the CHO cells are suitable for production of proteins used for human therapy. The expression of proteins in the cell is a complex mechanism, fundamentally depending on the DNA sequences in the cell nucleus. Secretion of translated proteins has been showed to be a bottleneck when improving expression. Secretion is initiated by the signal peptide, a n-terminal prolongation of the protein that is recognized by a signal recognition particle (SRP) when being translated by the ribosome. The sequence and structure of the signal peptide has been proved to affect secretion and altering the signal peptide could improve secretion even when changing signal peptide between different species. Designing variants of the signal peptides and analyzing protein expression might lead to improvements of the construct design and more protein produced from the cells, which would save time, money and material for the producer. To construct plasmids containing the gene of interest (GOI) and different signal peptides, several gene cloning methods were used. The plasmids were amplified using Escherichia coli (E. coli) transformation. The constructs were expressed by transfection into the CHO cell genome, and expression were analyzed using flow cytometry. When analyzing expression of a Fc-fusion protein with 5 different signal peptides, the signal peptide Azurocidin is the one showing highest expression levels in this study. In addition, IgG kapa and Albumin signal peptides did not show as high protein expression levels, even if they were better than the L1d and H5b signal peptides. Since signal peptides are exchangeable between proteins and species, it might be that Azurocidin is improving secretion and protein expression with other proteins than Fc-fusion proteins which would be an interesting aspect for further studies. When altering signal peptides with library sequences, the experimental challenges were crucial for the protein expression results and due to these issues, no library sequence could be seen to conquer others when it comes to protein expression levels. Transfection and cultivation procedures needs to be studied and improved before being able to draw conclusions about which signal peptide library sequences that might improve secretion and increase the protein expression.

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Maho, Maud. "Evaluation des effets des traitements par Rituximab versus corticothérapie seule sur la réponse auto-réactive des patients atteints de pemphigus. First-line Rituximab combined with short-term Prednisone versus Prednisone alone for the treatment of Pemphigus (RITUX 3) : a prospective, multicentre, parallel-group, open-label randomised trial Risk factors for short-term relapse in patients with pemphigus treated by Rituximab as first-line therapy Rituximab and corticosteroid effect on Desmoglein specific B cells and T follicular helper cells in patients with Pemphigus Modifications or the transcriptomic profile of autoreactive B cells from pemphigus patients after treatment with Rituximab or standard corticosteroid regimen Long-term increase of Kcnn4 potassium channel surface expression on B cells in pemphigus patients after Rituximab treatment Rituximab is an effective treatment in patients with Pemphigus Vulgaris and demonstrates a steroid-sparing effect Modifications of the BAFF/BAFF-Receptor axis in patients with pemphigus treated with rituximab versus standard corticosteroids regimen. CD11C+ B cells are mainly memory cells prone to differentiate into antibody-secreting cells". Thesis, Normandie, 2019. http://www.theses.fr/2019NORMR132.

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Le pemphigus est une maladie auto-immune spécifique de la peau et des muqueuses provoqué par des auto-anticorps (Ac) spécifiques des desmogléines (Dsg) 1 ou 3. Ces Ac pathogéniques inhibent l'adhésion cellulaire des kératinocytes. Le pemphigus se déclenche par la conjonction d’événements rares impliquant l’émergence puis la coopération de lymphocytes B (LB) et de lymphocytes LT auto-réactifs dans un contexte génétique et environnemental particulier. Jusqu’à présent, la première ligne de traitement du pemphigus était constituée de fortes doses de corticoïdes, qui sont de puissants immunosupresseurs systèmiques. Le Rituximab (RTX), un Ac monoclonal chimérique anti-CD20, constitue une thérapeutique innovante aboutissant à l’élimination des LB. L’étude clinique RITUX 3 a été conçue pour évaluer l’efficacité et l’innocuité du traitement utilisant le RTX associé à une courte corticothérapie dans le traitement de première intention du pemphigus par rapport au traitement de référence par la corticothérapie standard (CS). Dans un premier temps, notre analyse clinico-biologique des patients après 24 mois a démontré que l’utilisation du RTX associé à de la prednisone à court terme en traitement de première intention chez les patients atteints de pemphigus foliacé et vulgaire modéré à sévère est à la fois plus efficace et mieux toléré que le traitement de référence par la prednisone seule (89% de patients versus 34%). Cette efficacité a été confortée à plus long terme après la reconstitution du répertoire lymphocytaire B avec un risque de rechute de 2% à 36 mois. La présence d’une forme sévère de pemphigus au diagnostic (PDAI ≥ 45) et d’un taux d’Ac anti-Dsg à 3 mois supérieur aux valeurs seuils (anti-Dsg1 ≥ 20 ou anti-Dsg3 ≥ 120) sont associés à un risque de rechute précoce de 50%. Ces deux facteurs prédictifs permettent d'identifier un sous-groupe de patients présentant un risque élevé de rechute nécessitant une perfusion d'entretien de RTX au 6ème mois. Dans un deuxième temps, nous avons étudié l’impact des traitements par RTX et par CS chez les patients atteints de pemphigus afin de mieux appréhender la réponse auto-immune. La caractérisation phénotypique des LB auto-réactifs et l’analyse de la fréquence des LB capables de sécréter des immunoglobulines (Ig)G anti-Dsg par une approche ELISPOT a permis d’établir que l’efficacité du traitement par RTX dans le pemphigus semble liée à l’élimination des LB mémoires CD27+IgG+ spécifiques des Dsg. Des LB auto-réactifs Dsg restent détectables après RTX suite à la reconstitution lymphocytaire B, mais ces LB ont un phénotype naïf et non commuté (IgM) et ne secrètent plus d’IgG. En revanche, la persistance des LB auto-réactifs capables de sécréter des IgG anti-Dsg après traitement par CS est certainement à l’origine des rechutes fréquentes. L’analyse de l’expression génique ciblée à l’échelle unicellulaire a démontré qu’initialement, les LB spécifiques des Dsg ont un profil pro-inflammatoire avec l’expression de trois gènes codant pour les interleukines (IL)-1β, IL-12p35 et IL-23p19 et pour le gène de l’IRF5 (Interferon regulatory factor 5) par rapport aux LB non auto-réactifs. Le RTX et la CS ont des effets différents sur l'expression de ces gènes mais les deux réduisent l’expression génique d’IL-1β qui semble jouer un rôle important dans la physiopathologie du pemphigus. Parallèlement, l’analyse transcriptomique puis protéique des LB isolés des patients en rémission complète ou incomplète 6 ans après l’étude RITUX 1 a mis en évidence une augmentation d'expression de KCNN4 (Potassium calcium-activated channel subfamily N member 4) à la surface des LB chez les patients atteints de pemphigus en rémission complète pouvant influencer la maturation des LB
Pemphigus is an autoimmune disease of the skin and mucous membranes caused by autoantibodies (Ab) specific to desmoglein (Dsg) 1 or 3. These pathogenic Ab inhibit cell adhesion of keratinocytes. The development of pemphigus is associated with the conjunction of many uncommon events involving the emergence and then the cooperation of auto-reactive B cells and T cells link to genetic and environmental factors. Until now, the first line of treatment consisted of high doses of corticosteroids. Rituximab (RTX), an anti-CD20 chimeric monoclonal antibody, is an innovative therapy that results in B cells depletion. The RITUX 3 clinical trial was designed to evaluate the efficacy and safety of RTX combined with a short-course glucocorticoid therapy as a first-line treatment of pemphigus versus the standard treatment with standard corticosteroids (CS). As a first step, our clinico-biological analysis of patients after 24 months has shown that the use of RTX combined with short-term prednisone as a first-line treatment in patients with moderate to severe pemphigus is both more effective and better tolerated than the reference treatment with prednisone alone. Respectively, 89% of patients versus 34% in each group and both pemphigus foliaceus and pemphigus vulgaris patients responded. This efficacy was confirmed in the longer term after reconstitution of the B lymphocyte repertoire with a risk of relapse of only 2% at 36 months. The presence of a severe form of pemphigus at diagnosis (PDAI ≥ 45) and an anti-Dsg Ab level at 3 months above threshold values (anti-DSG1 ≥ 20 or anti-DSG3 ≥ 120) are associated with 50% risk of early relapse. These two predictive factors make it possible to identify a subgroup of patients at high risk of relapse requiring a maintenance infusion of RTX at the 6th month. In a second step, we studied the impact of RTX and CS treatments in patients with pemphigus in order to better understand the autoimmune response. The phenotypic characterization of auto-reactive B cells and the analysis of the frequency of B cells able of secreting anti-Dsg immunoglobulin (Ig) G by an ELISPOT approach demonstrated that the efficacy of RTX treatment in pemphigus seems related to the elimination of IgG-switched Dsg memory B-cells. Dsg specific B cells remain detectable after RTX when B cells return, but these B cells have a naïve and non-switched (IgM) phenotype and no longer secrete IgG. On the other hand, the persistence of self-reactive Dsg B cells capable of secreting IgG anti-Dsg after treatment with CS is certainly at the origin of the frequency of relapses. The unicellular targeted gene expression analysis demonstrated that initially, Dsg-specific B cells have a pro-inflammatory profile with the overexpression of three genes encoding Interleukin (IL) -1β, IL-12p35 and IL-23p19 and for the IRF5 gene (Interferon regulatory factor 5) compared to non-self-reactive B cells. RTX and CS have different effects on the expression of these genes, but both reduce the gene expression of IL-1β, which seems to play an important role in the pathophysiology of pemphigus

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Capítulos de libros sobre el tema "Antibody Secreting Cell"

Hsu, Myat Noe, Zirui Matthew Tay, Weikang Nicholas Lin y Shih-Chung Wei. "Screening of Antigen-Specific Antibody-Secreting Cells". En Handbook of Single Cell Technologies, 1–23. Singapore: Springer Singapore, 2020. http://dx.doi.org/10.1007/978-981-10-4857-9_27-1.

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Hsu, Myat Noe, Zirui Matthew Tay, Weikang Nicholas Lin y Shih-Chung Wei. "Screening of Antigen-Specific Antibody-Secreting Cells". En Handbook of Single-Cell Technologies, 471–93. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-10-8953-4_27.

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Karulin, Alexey Y., Melinda Katona, Zoltán Megyesi, Greg A. Kirchenbaum y Paul V. Lehmann. "Artificial Intelligence-Based Counting Algorithm Enables Accurate and Detailed Analysis of the Broad Spectrum of Spot Morphologies Observed in Antigen-Specific B-Cell ELISPOT and FluoroSpot Assays". En Methods in Molecular Biology, 59–85. New York, NY: Springer US, 2024. http://dx.doi.org/10.1007/978-1-0716-3690-9_5.

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AbstractAntigen-specific B-cell ELISPOT and multicolor FluoroSpot assays, in which the membrane-bound antigen itself serves as the capture reagent for the antibodies that B cells secrete, inherently result in a broad range of spot sizes and intensities. The diversity of secretory footprint morphologies reflects the polyclonal nature of the antigen-specific B cell repertoire, with individual antibody-secreting B cells in the test sample differing in their affinity for the antigen, fine epitope specificity, and activation/secretion kinetics. To account for these heterogeneous spot morphologies, and to eliminate the need for setting up subjective counting parameters well-by-well, CTL introduces here its cutting-edge deep learning-based IntelliCount™ algorithm within the ImmunoSpot® Studio Software Suite, which integrates CTL’s proprietary deep neural network. Here, we report detailed analyses of spots with a broad range of morphologies that were challenging to analyze using standard parameter-based counting approaches. IntelliCount™, especially in conjunction with high dynamic range (HDR) imaging, permits the extraction of accurate, high-content information of such spots, as required for assessing the affinity distribution of an antigen-specific memory B-cell repertoire ex vivo. IntelliCount™ also extends the range in which the number of antibody-secreting B cells plated and spots detected follow a linear function; that is, in which the frequencies of antigen-specific B cells can be accurately established. Introducing high-content analysis of secretory footprints in B-cell ELISPOT/FluoroSpot assays, therefore, fundamentally enhances the depth in which an antigen-specific B-cell repertoire can be studied using freshly isolated or cryopreserved primary cell material, such as peripheral blood mononuclear cells.

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Becza, Noémi, Zhigang Liu, Jack Chepke, Xing-Huang Gao, Paul V. Lehmann y Greg A. Kirchenbaum. "Assessing the Affinity Spectrum of the Antigen-Specific B Cell Repertoire via ImmunoSpot®". En Methods in Molecular Biology, 211–39. New York, NY: Springer US, 2024. http://dx.doi.org/10.1007/978-1-0716-3690-9_13.

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AbstractThe affinity distribution of the antigen-specific memory B cell (Bmem) repertoire in the body is a critical variable that defines an individual’s ability to rapidly generate high-affinity protective antibody specificities. Detailed measurement of antibody affinity so far has largely been confined to studies of monoclonal antibodies (mAbs) and are laborious since each individual mAb needs to be evaluated in isolation. Here, we introduce two variants of the B cell ImmunoSpot® assay that are suitable for simultaneously assessing the affinity distribution of hundreds of individual B cells within a test sample at single-cell resolution using relatively little labor and with high-throughput capacity. First, we experimentally validated that both ImmunoSpot® assay variants are suitable for establishing functional affinity hierarchies using B cell hybridoma lines as model antibody-secreting cells (ASC), each producing mAb with known affinity for a defined antigen. We then leveraged both ImmunoSpot® variants for characterizing the affinity distribution of SARS-CoV-2 Spike-specific ASC in PBMC following COVID-19 mRNA vaccination. Such ImmunoSpot® assays promise to offer tremendous value for future B cell immune monitoring efforts, owing to their ease of implementation, applicability to essentially any antigenic system, economy of PBMC utilization, high-throughput capacity, and suitability for regulated testing.

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Gaa, Ramona, Qingyong Ji y Achim Doerner. "Antibody-Secreting Cell Isolation from Different Species for Microfluidic Antibody Hit Discovery". En Methods in Molecular Biology, 313–25. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-3279-6_17.

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Kitikoon, Pravina, Crystal L. Loving y Amy L. Vincent. "Antibody Secreting Cell Assay for Influenza A Virus in Swine". En Methods in Molecular Biology, 347–53. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-0758-8_29.

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Florence, CHUA, OH Steve Kah Weng, YAP Miranda y TEO Wah Koon. "eRDF Medium Enhances Antibody Production of IgG and IgM Secreting Hybridomas." En Animal Cell Technology: Basic & Applied Aspects, 391–96. Dordrecht: Springer Netherlands, 1993. http://dx.doi.org/10.1007/978-94-011-2044-9_53.

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Shah, Hemangi B. y Kristi A. Koelsch. "B-Cell ELISPOT: For the Identification of Antigen-Specific Antibody-Secreting Cells". En Methods in Molecular Biology, 419–26. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2694-7_42.

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Krungkasem, C., H. Tachibana y S. Shirahata. "Identification of an Antibody-Secreting Human B Cell Line Undergoing “Light Chain Shifting”". En Animal Cell Technology: Basic & Applied Aspects, 639–43. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-011-5746-9_104.

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Yao, Lingling, Noémi Becza, Andrea Maul-Pavicic, Jack Chepke, Greg A. Kirchenbaum y Paul V. Lehmann. "Four-Color ImmunoSpot® Assays Requiring Only 1–3 mL of Blood Permit Precise Frequency Measurements of Antigen-Specific B Cells-Secreting Immunoglobulins of All Four Classes and Subclasses". En Methods in Molecular Biology, 251–72. New York, NY: Springer US, 2024. http://dx.doi.org/10.1007/978-1-0716-3690-9_15.

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AbstractThe B lymphocyte response can encompass four immunoglobulin (Ig) classes and four IgG subclasses, each contributing fundamentally different effector functions. Production of the appropriate Ig class/subclass is critical for both successful host defense and avoidance of immunopathology. The assessment of an antigen-specific B cell response, including its magnitude and Ig class/subclass composition, is most often confined to the antibodies present in serum and other biological fluids and neglects monitoring of the memory B cell (Bmem) compartment capable of mounting a faster and more efficient antibody response following antigen reencounter. Here, we describe how the frequency and Ig class and IgG subclass use of an antigen-specific Bmem repertoire can be determined with relatively little labor and cost, requiring only 8 × 105 freshly isolated peripheral blood mononuclear cells (PBMC), or if additional cryopreservation and polyclonal stimulation is necessary, 3 × 106 PBMC per antigen. To experimentally validate such cell saving assays, we have documented that frequency measurements of antibody-secreting cells (ASC) yield results indistinguishable from those of enzymatic (ELISPOT) or fluorescent (FluoroSpot) versions of the ImmunoSpot® assay, including when the latter are detected in alternative fluorescent channels. Moreover, we have shown that frequency calculations that are based on linear regression analysis of serial PBMC dilutions using a single well per dilution step are as accurate as those performed using replicate wells. Collectively, our data highlight the capacity of multiplexed B cell FluoroSpot assays in conjunction with serial dilutions to significantly reduce the PBMC requirement for detailed assessment of antigen-specific B cells. The protocols presented here allow GLP-compliant high-throughput measurements which should help to introduce high-dimensional Bmem characterization into the standard immune monitoring repertoire.

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Actas de conferencias sobre el tema "Antibody Secreting Cell"

Moffat, E. H., R. H. Furlong, A. L. Bloom y J. C. Giddings. "A MURINE MODEL FOR FACTOR VIII ANTIBODY ANTI-IDIOTYPE REAGENTS". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644030.

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The regulation of factor VIII antibody (FVIIIAb) production in haemophilic and non-haemophilic patients may be effected by anti-idiotype (Aid) antibodies which specifically react with FVIIIAb. Aid antibodies (reagents) were prepared from rabbits immunised with murine monoclonal FVIIIAb. Immuno fluorescent microscopy and cell culture studies were performed using murine hybridoma cells which secreted the FVIIIAbs.Immuno fluorescence studies examined the ability of the Aid reagents to bind to acetone fixed FVIIIAb secreting hybridoma cells. Positive surface membrane and intra-cytoplasmic staining patterns were seen with the Aid reagent when incubated with the corresponding murine hybridoma cell line. This reaction was blocked subsequent to the addition of the corresponding monoclonal FVIIIAb but was preserved when unrelated monoclonal FVIIIAb was incubated with the hybridoma cells. No fluorescence was observed when Aid reagent was incubated with unrelated FVIIIAb secreting hybridoma cultures.Following the addition of Aid reagent to FVIIIAb secreting hybridoma cultures and incubation for 19 hours, the resultant hybridoma supernatants were examined for FVIIIAb content using an immunoradiometric technique. The Aid reagent failed to inhibit FVIIIAb secretion by hybridoma cells. Thus, although Aid reagents were capable of binding to fixed FVIIIAb secreting cells, they failed to inhibit FVIIIAb secretion from hybridoma cultures. The conjugation of Aid reagent with immunotoxin may however have cytotoxic potential. The murine model provides a method for the study of Aid regulation of FVIIIAb production in haemophilia.

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Romanuik, Sean F., Samantha M. Grist, Moeed Haq, Bonnie L. Gray, Naveed Gulzar y Jamie K. Scott. "The Microfluidic Trapping of Antibody-Secreting Cells". En ASME 2010 8th International Conference on Nanochannels, Microchannels, and Minichannels collocated with 3rd Joint US-European Fluids Engineering Summer Meeting. ASMEDC, 2010. http://dx.doi.org/10.1115/fedsm-icnmm2010-30845.

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Therapeutic antibodies (Abs) are a rapidly growing and economically promising biotechnological research area. Therapeutic Ab production typically involves screening large numbers of Ab-secreting cells (ASCs) in order to identify those producing Abs targeting a specific antigen (Ag) with the highest affinity; a process often requiring weeks to complete. We are contributing to a multidisciplinary project focused upon the development of an immunobiosensing array ultimately intended to directly monitor the Ag-specific Ab production by thousands of ASCs on a single slide in real-time. Each ASC shall be microfluidically guided and trapped near a surface plasmon (SP) resonant nanohole array sensor so as to detect the binding of secreted Abs to Ag immobilized onto the sensor’s surface. This paper presents the initial progress of our contribution to this project: the development of polymeric microfluidic devices to guide and trap large ASC populations within arrays of single-cell traps. More specifically, this paper presents several different polymer-based microfluidic trapping devices, based upon perfusive flow-through cell traps and microwells which trap settling cells, which have been evaluated using COMSOL® simulations and tested using microsphere- and cell-based flow experiments. Our initial results are promising, and verify the functionality of our microfluidic cell trap designs.

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Ramonell, R. P., M. Brown, M. C. Woodruff, J. M. Levy, S. K. Wise, J. DelGaudio, M. Duan et al. "Single Cell Analysis of Human Nasal Mucosal IgE Antibody Secreting Cells Reveals Newly Minted Phenotype". En American Thoracic Society 2023 International Conference, May 19-24, 2023 - Washington, DC. American Thoracic Society, 2023. http://dx.doi.org/10.1164/ajrccm-conference.2023.207.1_meetingabstracts.a6209.

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Walker, Natalie, Aiden Mclinden, Aiden Mclinden, Yuta Ibuki, Keizo Misumi y Vibha N. Lama. "Broncho-vascular mesenchymal stromal cells guided spatiotemporal establishment of antibody secreting cell pro-survival niches in the lung". En ERS Lung Science Conference 2022 abstracts. European Respiratory Society, 2022. http://dx.doi.org/10.1183/23120541.lsc-2022.100.

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Lee, Frances E. H., Jessica L. Halliley, Andrew P. Moscatiello, Ann R. Falsey, Edward E. Walsh y Ignacio Sanz. "Novel Method For Diagnosing Microbial Infections With A Blood Antibody Secreting Cell Assay: The “MicroBspot™"". En 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.a6845.

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Cheng, Chao, Ermin Xie, Xinyi Huang, Suo Chu, Xiaorui Chen, Bixia Lei, Zhenwei Zhong, Jiantao Wang, Huajing Wang Xian-Yang Li y Xiaowen He. "241 Dual targeting liver cancer by GPC3 CAR-T cells secreting a bispecific T cell engager antibody for an intracellular tumor antigen AFP". En SITC 38th Annual Meeting (SITC 2023) Abstracts. BMJ Publishing Group Ltd, 2023. http://dx.doi.org/10.1136/jitc-2023-sitc2023.0241.

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Mclinden, A. P., Y. Ibuki, N. M. Walker, I. Grigorova y V. N. Lama. "Novel Bronchovascular-Bundle-Associated Mesenchymal Stromal Cell Population Promote the Survival of Antibody Secreting Cells in Lung Allografts with a Restrictive Allograft Syndrome Phenotype". En American Thoracic Society 2022 International Conference, May 13-18, 2022 - San Francisco, CA. American Thoracic Society, 2022. http://dx.doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a3942.

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Van Lent, Julie, Iene Rutten, Karen Ven, Jolien Breukers, Eleonore Verstraete, Katrien Van, Julie Van et al. "MICROFLUIDIC TOOLS FOR STUDYING SINGLE CELL SECRETIONS: A CASE STUDY ON HYBRIDOMA ANTIBODY SECRETION". En The 28th International Conference on Miniaturized Systems for Chemistry and Life Sciences - Micro-Total Analysis Systems. San Diego: Chemical and Biological Microsystems Society, 2024. https://doi.org/10.70477/zhun7496.

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Baker, J. B., M. P. McGrogan, C. Simonsen, R. L. Gronke y B. W. Festoff. "STRUCTURE AND PROPERTIES OF PROTEASE NEXIN I". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644765.

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Human foreskin fibroblasts secrete several different serine protease inhibitors which differ in size and protease specificities. These proteins, called protease nexins (PNs) all form SDS-resistant complexes with their protease targets. Fibroblast surface receptors recognize the protease-PN complexes and mediate their delivery to lysosomes. PNI is a 45 kilodalton glycoprotein that rapidly inhibits several arg or lys-specific proteases including trypsin, thrombin, and urokinase (k assoc.∼ 4×l06,∼ 6×105 and ∼ 2×105, m−1s−1 respectively). Like antithrombin III, PNI binds heparin and inhibits thrombin at a vastly accelerated rate in the presence of this glycoaminoglycan. Immunofluorescence studies show that in addition to secreting PNI foreskin fibroblasts carry this inhibitor on their surfaces. PNI cDNA has been cloned and sequenced. A mixed oligonucleotide probe derived from PNI N-terminal sequence was used to probe a foreskin fibroblast cDNA library constructed with λGT10. Identification of PNI cDNAs has been verified by sequencing and by expressing active PNI protein in mammalian cells. The full amino acid sequence of PNI, deduced from cDNA sequencing, is 392 residues long and has 30% homology to antithrombin III. An arg-ser pair 32 residues from the C-terminus of the inhibitor is proposed as the reactive center P1-P1 residues. In the hinge region a lys residue is present in a position occupied by a ginor glu residue in other serpins. PNI mRNA exists in 2 slightly different forms:One (αPNI) yields a thr-arg-ser sequence wherethe other βPNI) yields a thr-thr-gly-ser sequence. The presence of the appropriate splice acceptor sites in the genome indicates that these forms are generated from a single gene by alternative splicing. Expressed aPNI and 0PNI proteins both bind thrombin and urokinase. In foreskin fibroblaststhe α form of PNI mRNA predominates over the β form by about 2:1. In foreskin fibroblast cultures secreted PNI inhibits the mitogenic response to thrombin and regulate secreted urokinase. Purified PNI added to human fibrosarcoma (HT1080) cells inhibitsthe tumor cell-mediated destruction of extracellular matrix and transiently, but dramatically, inhibits tumor cell growth. PNI or PNI-like inhibitors may function at multiple physiological sites. The β form of PNI is virtually identical to a glia-derived neurite promoting factor, the cDNA for which has been recently cloned and sequenced by Gloor et al (1). The neurite outgrowth activity of PNI may result from inhibition of a thrombin-like protease that is associated with neurons, since a number of thrombin inhibitors stimulate neurite extension. Recent immunofluoresence experiments, carried out with D. Hantai (Inserm; Paris) demonstrate that anti-PNI antibody intensely stains neuromuscular synapses. In addition, a PNI-like inhibitor is associated with platelets. At low (0.5 nM <) 125I-thrombin concentrations formation of 125I-thrombin-platelet PNI complexes accounts for most of the specific binding of 125I-thrombin to platelets (2). Although the platelet-associated form of PNI is electrophoretically and immunologically indistinguishable from fibroblast PNI, it does not bind urokinase, suggesting that it may be distinct.(1) Gloor, S., K. Odink, J. Guenther, H. Nick, and D. Monard. (1986) Cell 47:687-693.(2) Gronke, R.S., B.L. Bergman, and J.B. Baker. (1987) J. Biol. Chem. (in press)

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Grabowski, F. E. "RHEOLOGY AND PRIMARY HEMOSTASIS". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643986.

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Overview The adhesion-aggregation of platelets to a site of vessel wall injury is a quintessential blood flow phenomenon. Firstly, platelets are driven to the vicinity of the vessel wall by a form of convective diffusion in which red cells both mechanically augment the effective platelet diffusivity (Turitto et al., Ind. Eng. Chem. Fund. 11:216-223, 1972; Grabowski et al., Ind. Eng. Chem. Fund. 11:224-232, 1972) and enhance the near-wall piatelet concentration (Ti11es and Eckstein, Microvasc Res., In press, 1987). Secondly, red cells subjected to physiologic shear forces are capable of secreting sufficient adenine nucleotides to induce primary platelet aggregation without themselves undergoing frank lysis (Reimers et al, Blood 64:1200-1206, 1984). This "humoral" effect of erythrocytes is likely to contribute to primary hemostasis in a shear stress-dependent manner. Thirdly, endothelial cells are able to modulate platelet aggregation at a site of vessel injury by producing prostacyclin (and perhaps other antithrombotic substances) in a manner which increases with vessel shear rate (Grabowski et al, Blood 62:301a, 1983); production for a large range of arterial shear rates appears to be limited by plasma-borne substrate (arachidonate). This manner of production ensures a concentration of prostacyclin in the near-wall region which remains relatively independent of shear rate.Imaging primary hemostasis. In our work, epi-fluorescence videomicroscopy has allowed real time imaging of platelet adhesion-aggregation to a simulated vessel wall injury. The injury model is an endothelial cell monolayer (ECM) across which, prior to ECM exposure to flowing blood, a 6-0 sterile suture is drawn in a direction transverse to flow. Microinjuries result which measure 70 ± 15μm (Mean ± SD) in width. The fluorescent label is the TAB murine monoclonal antibody (courtesy of Dr. R.P. McEver) directed against human platelet GPIIB, together with a fluorescein-conjugated goat F(ab')2 against murine inmunoglobulin. The injured ECM's, grown to confluence on rectangular cover glasses precoated with microfibrillar collagen, comprise one wall of a flow chamber mounted on a vertical microscope stage. On microinjury sites and at shear rates of 100 to 700 sec-1, computer-enhanced video images show adherence, remodelling and growth of chains of platelet aggregates. Aligned with the flow direction, these chains have a spacing of approximately 30)im, a length similar to the average endothelial cell diameter. One may speculate that such chains provide a scaffold for wound healing insofar as they are likely rich in agents chemotactic for leukocytes and in platelet-derived growth factor.Modulatory role of endothelium. When the ECM's are pre treated with 1.0 mM FC lysine acetyl sal icy late (LA), aggregate length increases (P<0.001) up totwo-fold, outflow levels by RIA of serum thromboxane B2 increase (8 of 8 paired runs), and outflow levels of prostacyclin by RIA for 6-Keto PGFiot decrease (5 of 7 paired runs). The Table gives data for one of four similar experiments at 270 sec-1 and following five minutes of flow. These data imply that products of ECM which are inhibitable by aspirin modulate local adhesion-aggregation; their inhibition, as by vasculitis or drugs, may give rise to thrombotic states.Bleeding disorders. Aggregate length is reduced in von Willebrand's disease (4 patients), Hermansky-Pudlak syndrome (2 patients), and after 300 mg oral aspirin (Tablet 4 donors). The reduction in the first two, however, is greater (P<0.01) than that for oral aspirin. With oral aspirin, further, there is a paradoxic increase in the percent platelet coverage of the injury area. Summary. Rheology has profound effects on the rate, structure, and modulation of primary hemostasis. Many of these effects can be studied via real-time, epi-fluorescence videomicroscopy of platelet adhesion-aggregation to a site of injury to an endothelial cell monolayer exposed to flowing blood. The model described has application to the study of thrombotic and hemostatic disorders and unstable angina.

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Informes sobre el tema "Antibody Secreting Cell"

Baszler, Timothy, Igor Savitsky, Christopher Davies, Lauren Staska y Varda Shkap. Identification of bovine Neospora caninum cytotoxic T-lymphocyte epitopes for development of peptide-based vaccine. United States Department of Agriculture, marzo de 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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วีรกุล, ปราจีน, อัจฉริยา ไศละสูต, คณิศักดิ์ อรวีระกุล, วาสนา ภิญโญชนม์, สุดารัตน์ ดำรงค์วัฒนโภคิน y สันนิภา สุรทัตต์. การวิจัยและการพัฒนาวิธีวินิจฉัย ควบคุมและป้องกันโรคอหิวาต์สุกรในประเทศไทย : รายงานการวิจัย. จุฬาลงกรณ์มหาวิทยาลัย ; สำนักงานคณะกรรมการวิจัยแห่งชาติ, 2002. https://doi.org/10.58837/chula.res.2002.61.

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ชุดโครงการวิจัยนี้ แบ่งการศึกษาออกเป็น 4 โครงการย่อย ซึ่งประกอบด้วย โครงการที่ 1 การศึกษาเปรียบเทียบระดับแอนติบอดีในแม่สุกรที่ได้รับการฉีดวัคซีนโปรแกรม 3 ชนิด และผลกระทบของภูมิคุ้มกันถ่ายทอดจากแม่สู่ลูกในการสร้างภูมิคุ้มกันในลูกสุกรเมื่อได้รับการฉีดวัคซีน โครงการที่ 2 การพัฒนาวิธีการตรวจวินิจฉัยโรคอหิวาต์สุกรและการพัฒนาการตรวจวัดภูมิคุ้มกันชนิดเซลล์ต่อเชื้อไวรัสอหิวาต์สุกร โครงการที่ 3 การศึกษาพัฒนาวิธีผลิตวัคซีนอหิวาต์สุกรชนิดเซลล์เพาะเลี้ยง และโครงการที่ 4 การศึกษาระดับภูมิคุ้มกันถ่ายทอดที่สามารถป้องกันการเกิดอหิวาต์สุกรเมื่อได้รับการฉีดเชื้อพิษอหิวาต์สุกร Chiangmai/98 โครงการที่ 1 การศึกษาสถานภาพของระดับแอนติบอดีในแม่สุกรที่ได้รับการฉีดวัคซีนป้องกันโรคอหิวาต์สุกร 3 โปรแกรมพบว่า ระดับแอนติบอดีในแม่สุกรที่ 1 สัปดาห์หลังคลอดในโปรแกรมที่ได้รับการฉีดวัคซีน 3 สัปดาห์ก่อนคลอดหรือโปรแกรมฉีดพร้อมกันทุกตัวในฟาร์มปีละ 3 ครั้ง ไม่มีความแตกต่างกัน ส่วนแม่สุกรที่ฉีดวัคซีนโปรแกรม 3 สัปดาห์หลังคลอดมีค่าใช้จ่ายของระดับแอนติบอดีต่ำกว่าทั้ง 2 โปรแกรมอย่างมีนัยสำคัญทางสถิติ (p<0.05) ผลการศึกษาระดับภูมิคุ้มกันถ่ายทอดสู่ลูกสุกรที่เกิดจากแม่ที่ได้รับการฉีดวัคซีนทั้ง 3 โปรแกรมเมื่อลูกสุกรอายุ 1 และ 3 สัปดาห์ มีระดับแอนติบอดีที่สอดคล้องกับแอนติบอดีในแม่ การศึกษาผลกระทบของระดับภูมิคุ้มกันถ่ายทอดจากแม่สู่ลูกที่อาจมีผลรบกวนการสร้างภูมิคุ้มกันในลูกสุกรเมื่อฉีดวัคซีน ผลการศึกษาการประเมินประสิทธิภาพของโปรแกรมการฉีดวัคซีนในลูกสุกรครั้งเดียวที่อายุ 3 สัปดาห์หรือ 2 ครั้งที่อายุ 3 และ 5 สัปดาห์ พบว่า วัคซีนให้ความคุ้มโรคได้ แม้ลูกสุกรมีภูมิคุ้มกันถ่ายทอดจากแม่ในระดับที่แตกต่างกัน ณ วันฉีดวัคซีน อย่างไรก็ตามพบว่าภูมิคุ้มกันถ่ายทอดในระดับสูงจะรบกวนการสร้างภูมิคุ้มกันชนิดเซลล์ในลูกสุกรเมื่อฉีดวัคซีน โครงการที่ 2 การตรวจวินิจฉัยโรคอหิวาต์สุกรโดยวิธี reverse transcriptase polymerase chain reaction (RT-PCR) พบว่าเมื่อใช้ primer 324 และ 326 เป็นวิธีที่มีความไวและความจำเพาะสูง สามารถตรวจหาสารพันธุกรรมของเชื้อไวรัสอหิวาต์สุกรที่แยกได้ในประเทศทุก genogroups และให้ผลสอดคล้องกับวิธีการแยกและพิสูจน์เชื้อในเซลล์เพาะเลี้ยง จึงเหมาะสำหรับนำมาใช้ในการวินิจฉัยโรคอหิวาต์สุกรในประเทศ (โครงการที่ 2.1) สำหรับวิธี RT-PCR เพื่อตรวจหาสารพันธุกรรมของไวรัสอหิวาต์สุกรในส่วนของ gp 55 ร่วมกับการใช้เอ็นไซม์จำเพาะ เพื่อแยกไวรัสจากวัคซีนที่แยกได้จากท้องที่ พบว่ามีข้อจำกัดในเรื่องความไว และความจำเพาะ จึงอาจไม่เหมาะสำหรับนำมาใช้ในการวินิจฉัยโรคและแยกแยะชนิดของเชื้อในประเทศ การพัฒนาวิธีการตรวจวัดภูมิคุ้มกันชนิดเซลล์โดยการตรวจหาปริมาณเซลล์ที่สร้างแอนติบอดี (antibody secreting cells) และเซลล์ที่สร้างอินเตอเฟอรอนแกมม่า (IFN[gamma]) ที่จำเพาะต่อเชื้ออหิวาต์สุกรเพื่อใช้เป็นตัวบ่งชี้ระดับ antiviral immunity และใช้ในการประเมินประสิทธิภาพของวัคซีนอหิวาต์สุกร ผลการวิจัยสามารถพัฒนาเทคนิค ELISPOT เพื่อตรวจวัดระดับภูมิคุ้มกันชนิดเซลล์ต่อเชื้ออหิวาต์สุกรได้เป็นผลสำเร็จ (โครงการที่ 2.2) โครงการที่ 3 พัฒนาวิธีผลิตวัคซีนอหิวาต์สุกรเชื้อเป็นเซลล์เพาะเลี้ยงโดยใช้เชื้อไวรัสวัคซีนสเตรน GPE เพาะเลี้ยงในเซลล์ FS-L[subscript 3] พบว่าสามารถนำมาใช้ผลิตเป็นวัคซีนชนิดดูดแห้งที่ได้มาตรฐานทางห้องปฏิบัติการเป็นวัคซีนที่มีความปลอดภัยสูงและให้ความคุ้มครองได้อย่างสมบูรณ์ในสุกรเมื่อฉีดพิษทับวัคซีนที่ผลิตได้มีความเหมาะสมที่จะนำไปผลิตในเชิงอุตสาหกรรมต่อไป โครงการที่ 4 การทดสอบภูมิคุ้มกันถ่ายทอดจากแม่และการฉีดวัคซีนในลูกสุกรต่อไวรัสอหิวาต์สุกร Chiangmai/98 ซึ่งเป็นเชื้อใน genogroup 2.2 พบว่าภูมิคุ้มกันถ่ายทอดมีผลน้อยมากต่อการให้ความคุ้มครองโรคต่อการฉีดพิษทับด้วยเชื้อ genogroup 2.2 นอกจากนั้นภูมิคุ้มกันถ่ายทอดมีผลรบกวนการสร้างภูมิคุ้มกันทั้งชนิดเซลล์และแอนติบอดีอย่างมีนัยสำคัญทางสถิติ (p<0.05) ต่อการฉีดวัคซีน

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