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single b cell cloning

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https://www.readbyqxmd.com/read/29036679/human-recombinant-fab-fragment-from-combinatorial-libraries-of-a-b-cell-lymphoma-patient-recognizes-core-protein-of-chondroitin-sulfate-proteoglycan-4
#1
Yoko Egami, Yuta Narushima, Motohiro Ohshima, Akira Yoshida, Naruki Yoneta, Yasufumi Masaki, Kunihiko Itoh
CD antigens are well known as therapeutic targets of B-cell lymphoma. To isolate therapeutic antibodies that recognize novel targets other than CD antigens, we constructed a phage display combinatorial antibody Fab library from bone marrow lymphocytes of B-cell lymphoma patient. To eliminate antibodies reactive with known B-cell lymphoma antigen, non-hematopoietic and patient's sera reactive HeLaS3 cells was selected as a target of whole cell panning. Five rounds of panning against live HeLaS3 cells retrieved single Fab clone, termed AHSA (Antibody to HeLa Surface Antigen)...
September 28, 2017: Journal of Biochemistry
https://www.readbyqxmd.com/read/29027080/different-expression-systems-resulted-in-varied-binding-properties-of-anti-paclitaxel-single-chain-variable-fragment-antibody-clone-1c2
#2
Gorawit Yusakul, Seiichi Sakamoto, Poomraphie Nuntawong, Hiroyuki Tanaka, Satoshi Morimoto
The binding properties of recombinant antibody fragments, such as affinity and specificity, determine their usefulness for therapeutic and analytical applications. Anti-paclitaxel single-chain variable fragment clone 1C2 (anti-PT scFv1C2) was expressed using Escherichia coli cell and Bombyx mori larvae expression systems. The binding characteristics of the scFvs were evaluated using indirect competitive ELISA. The linear range of binding between anti-PT scFv1C2 and paclitaxel was significantly different between the anti-PT scFv1C2s derived from E...
October 12, 2017: Journal of Natural Medicines
https://www.readbyqxmd.com/read/28981169/mir-200c-3p-spreads-invasive-capacity-in-human-oral-squamous-cell-carcinoma-microenvironment
#3
Tomoyo Kawakubo-Yasukochi, Masahiko Morioka, Mai Hazekawa, Atsushi Yasukochi, Takuya Nishinakagawa, Kazuhiko Ono, Shintaro Kawano, Seiji Nakamura, Manabu Nakashima
Oral squamous cell carcinoma (OSCC) constitutes over 90% of all cancers in the oral cavity. The prognosis for patients with invasive OSCC is poor; therefore, it is important to understand the molecular mechanisms of invasion and subsequent metastasis not only to prevent cancer progression but also to detect new therapeutic targets against OSCC. Recently, extracellular vesicles-particularly exosomes-have been recognized as intercellular communicators in the tumor microenvironment. As exosomic cargo, deregulated microRNAs (miRNAs) can shape the surrounding microenvironment in a cancer-dependent manner...
October 5, 2017: Molecular Carcinogenesis
https://www.readbyqxmd.com/read/28971969/natural-polyreactive-iga-antibodies-coat-the-intestinal-microbiota
#4
Jeffrey J Bunker, Steven A Erickson, Theodore M Flynn, Carole Henry, Jason C Koval, Marlies Meisel, Bana Jabri, Dionysios A Antonopoulos, Patrick C Wilson, Albert Bendelac
Large quantities of immunoglobulin A (IgA) are constitutively secreted by intestinal plasma cells to coat and contain the commensal microbiota, yet the specificity of these antibodies remains elusive. Here, we profiled the reactivities of single murine IgA plasma cells by cloning and characterizing large numbers of monoclonal antibodies. IgAs were not specific to individual bacterial taxa but rather polyreactive, with broad reactivity to a diverse but defined subset of microbiota. These antibodies arose at low frequencies among naïve B cells, and were selected into the IgA repertoire upon recirculation in Peyer's patches...
September 28, 2017: Science
https://www.readbyqxmd.com/read/28965920/selection-and-validation-of-antibody-clones-against-igg-and-iga-subclasses-in-switched-memory-b-cells-and-plasma-cells
#5
Elena Blanco, Martin Perez-Andres, Luzalba Sanoja-Flores, Marjolein Wentink, Ondrej Pelak, Marta Martín-Ayuso, Georgiana Grigore, Juan Torres-Cañizales, Eduardo López-Granados, Tomas Kalina, Mirjam Van der Burg, Sonia Arriba-Méndez, Santiago Santa Cruz, Noemí Puig, Jacques J M van Dongen, Alberto Orfao
The clinical value of assessing immunoglobulin (Ig)G and IgA subclasses in addition to the isotypes of soluble Igs in serum has been well established. >20years ago, the International Union of Immunological Societies and the World Health Organization performed collaborative studies in order to validate antibody (Ab) clones for the detection of IgG and IgA subclasses for a broad range of laboratory assays, but except for flow cytometry. Here we analyzed the performance of commercially available Ab clones to detect IgG and IgA subclasses in memory B-cells and plasma cells (PCs) by flow cytometry...
September 28, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28951589/molecular-imaging-using-an-anti-human-tissue-factor-monoclonal-antibody-in-an-orthotopic-glioma-xenograft-model
#6
Hiroki Takashima, Atsushi B Tsuji, Tsuneo Saga, Masahiro Yasunaga, Yoshikatsu Koga, Jun-Ichiro Kuroda, Shigetoshi Yano, Jun-Ichi Kuratsu, Yasuhiro Matsumura
Nuclear medicine examinations for imaging gliomas have been introduced into clinical practice to evaluate the grade of malignancy and determine sampling locations for biopsies. However, these modalities have some limitations. Tissue factor (TF) is overexpressed in various types of cancers, including gliomas. We thus generated an anti-human TF monoclonal antibody (mAb) clone 1849. In the present study, immunohistochemistry performed on glioma specimens using anti-TF 1849 mAb showed that TF expression in gliomas increased in proportion to the grade of malignancy based on the World Health Organization (WHO) classification, and TF was remarkably expressed in necrosis and pseudopalisading cells, the histopathological hallmarks of glioblastoma multiforme (GBM)...
September 26, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28923441/car-t-cell-immunotherapy-in-hematology-and-beyond
#7
Claudia Rossig
Chimeric T cell receptors (CARs) combine extracellular antigen recognition domains and T cell activation components in single molecules. CAR gene transfer thereby allows to generate T cells with engineered specificities. The translational development of CAR-based T cell therapies is most advanced in B cell cancers where CAR-engineered T cells against the B lineage antigen CD19 have generated impressive results in early clinical trials. CARs are now also explored as tools to eliminate autoreactive B cell clones and to engineer T cells with immunosuppressive function for preventing pathological auto- or alloresponses...
September 18, 2017: Clinical Immunology: the Official Journal of the Clinical Immunology Society
https://www.readbyqxmd.com/read/28903565/liquid-biopsies-the-clinics-and-the-molecules
#8
V Kubaczková, L Sedlarikova, B Bollová, V Sandecká, M Stork, L Pour, S Sevcikova
Unlike bone marrow biopsies, liquid biopsies represent a gentler, more accessible, less painful, repeatable and more comprehensive approach to get biologically relevant information about the entire tumor but also about treatment response and level of minimal residual disease. This is all possible since peripheral blood contains not only circulating tumor cells but also many circulating molecules of nucleic acids (microRNA, cell-free DNA, long non-coding RNA etc.). Multiple myeloma is a genetically heterogeneous disease characterized by multifocal tumor deposits in the bone marrow but also focal lesions elsewhere...
2017: Klinická Onkologie: Casopis Ceské a Slovenské Onkologické Spolecnosti
https://www.readbyqxmd.com/read/28890334/crispr-mediated-base-editing-enables-efficient-disruption-of-eukaryotic-genes-through-induction-of-stop-codons
#9
Pierre Billon, Eric E Bryant, Sarah A Joseph, Tarun S Nambiar, Samuel B Hayward, Rodney Rothstein, Alberto Ciccia
Standard CRISPR-mediated gene disruption strategies rely on Cas9-induced DNA double-strand breaks (DSBs). Here, we show that CRISPR-dependent base editing efficiently inactivates genes by precisely converting four codons (CAA, CAG, CGA, and TGG) into STOP codons without DSB formation. To facilitate gene inactivation by induction of STOP codons (iSTOP), we provide access to a database of over 3.4 million single guide RNAs (sgRNAs) for iSTOP (sgSTOPs) targeting 97%-99% of genes in eight eukaryotic species, and we describe a restriction fragment length polymorphism (RFLP) assay that allows the rapid detection of iSTOP-mediated editing in cell populations and clones...
September 21, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28885603/functional-characterization-of-selected-universal-stress-protein-from-salvia-miltiorrhiza-smusp-in-escherichia-coli
#10
Xiao-Fan Wang, Jiao Su, Na Yang, Hui Zhang, Xiao-Yan Cao, Jie-Fang Kang
The multigene universal stress protein (USP) family is evolutionarily conserved. Members play indispensable roles in plant tolerance to abiotic stresses. Although relatively well-characterized in model plants, such as Arabidopsis thaliana and Oryzasativa, this family has not been investigated in Salvia miltiorrhiza, an important herbal plant for which yields can be limited by various abiotic stresses. Here, we identified 32 USP family members in the S. miltiorrhiza genome, and used phylogenetic analysis to sort these SmUSPs into four groups...
September 8, 2017: Genes
https://www.readbyqxmd.com/read/28884285/immune-tb-antibody-phage-display-library-as-a-tool-to-study-b-cell-immunity-in-tb-infections
#11
Nurul Hamizah Hamidon, Siti Suraiya, Maria E Sarmiento, Armando Acosta, Mohd Nor Norazmi, Theam Soon Lim
B cells and in particular antibodies has always played second fiddle to cellular immunity in regard to tuberculosis (TB). However, recent studies has helped position humoral immunity especially antibodies back into the foray in relation to TB immunity. Therefore, the ability to correlate the natural antibody responses of infected individuals toward TB antigens would help strengthen this concept. Phage display is an intriguing approach that can be utilized to study antibody-mediated responses against a particular infection via harvesting the B cell repertoire from infected individuals...
September 7, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28878350/functional-association-of-cellular-microtubules-with-viral-capsid-assembly-supports-efficient-hepatitis-b-virus-replication
#12
Masashi Iwamoto, Dawei Cai, Masaya Sugiyama, Ryosuke Suzuki, Hideki Aizaki, Akihide Ryo, Naoko Ohtani, Yasuhito Tanaka, Masashi Mizokami, Takaji Wakita, Haitao Guo, Koichi Watashi
Viruses exploit host factors and environment for their efficient replication. The virus-host interaction mechanisms for achieving an optimal hepatitis B virus (HBV) replication have been largely unknown. Here, a single cell cloning revealed that HepAD38 cells, a widely-used HBV-inducible cell line, contain cell clones with diverse permissiveness to HBV replication. The HBV permissiveness was impaired upon treatment with microtubule inhibitor nocodazole, which was identified as an HBV replication inhibitor from a pharmacological screening...
September 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28841417/clonal-evolution-of-autoreactive-germinal-centers
#13
Søren E Degn, Cees E van der Poel, Daniel J Firl, Burcu Ayoglu, Fahd A Al Qureshah, Goran Bajic, Luka Mesin, Claude-Agnès Reynaud, Jean-Claude Weill, Paul J Utz, Gabriel D Victora, Michael C Carroll
Germinal centers (GCs) are the primary sites of clonal B cell expansion and affinity maturation, directing the production of high-affinity antibodies. This response is a central driver of pathogenesis in autoimmune diseases, such as systemic lupus erythematosus (SLE), but the natural history of autoreactive GCs remains unclear. Here, we present a novel mouse model where the presence of a single autoreactive B cell clone drives the TLR7-dependent activation, expansion, and differentiation of other autoreactive B cells in spontaneous GCs...
August 24, 2017: Cell
https://www.readbyqxmd.com/read/28835708/exploring-dengue-genome-to-construct-a-multi-epitope-based-subunit-vaccine-by-utilizing-immunoinformatics-approach-to-battle-against-dengue-infection
#14
Mudassar Ali, Rajan Kumar Pandey, Nazia Khatoon, Aruna Narula, Amit Mishra, Vijay Kumar Prajapati
Dengue is considered as a major health issue which causes a number of deaths worldwide each year; tropical countries are majorly affected by dengue outbreaks. It is considered as life threatening issue because, since many decades not a single effective approach for treatment and prevention of dengue has been developed. Therefore, to find new preventive measure, we used immunoinformatics approaches to develop a multi-epitope based subunit vaccine for dengue which can generate various immune responses inside the host...
August 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28813413/polylox-barcoding-reveals-haematopoietic-stem-cell-fates-realized-in-vivo
#15
Weike Pei, Thorsten B Feyerabend, Jens Rössler, Xi Wang, Daniel Postrach, Katrin Busch, Immanuel Rode, Kay Klapproth, Nikolaus Dietlein, Claudia Quedenau, Wei Chen, Sascha Sauer, Stephan Wolf, Thomas Höfer, Hans-Reimer Rodewald
Developmental deconvolution of complex organs and tissues at the level of individual cells remains challenging. Non-invasive genetic fate mapping has been widely used, but the low number of distinct fluorescent marker proteins limits its resolution. Much higher numbers of cell markers have been generated using viral integration sites, viral barcodes, and strategies based on transposons and CRISPR-Cas9 genome editing; however, temporal and tissue-specific induction of barcodes in situ has not been achieved. Here we report the development of an artificial DNA recombination locus (termed Polylox) that enables broadly applicable endogenous barcoding based on the Cre-loxP recombination system...
August 24, 2017: Nature
https://www.readbyqxmd.com/read/28768640/clonal-expansion-and-interrelatedness-of-distinct-b-lineage-compartments-in-multiple-myeloma-bone-marrow
#16
Leo Hansmann, Arnold Han, Livius Penter, Michaela Liedtke, Mark M Davis
Multiple myeloma is characterized by the clonal expansion of malignant plasma cells in the bone marrow. But the phenotypic diversity and the contribution of less predominant B-lineage clones to the biology of this disease have been controversial. Here, we asked whether cells bearing the dominant multiple myeloma immunoglobulin rearrangement occupy phenotypic compartments other than that of plasma cells. To accomplish this, we combined 13-parameter FACS index sorting and t-Stochastic Neighbor Embedding (t-SNE) visualization with high-throughput single-cell immunoglobulin sequencing to track selected B-lineage clones across different stages of human B-cell development...
September 2017: Cancer Immunology Research
https://www.readbyqxmd.com/read/28717136/association-of-plasma-and-urinary-mutant-dna-with-clinical-outcomes-in-muscle-invasive-bladder-cancer
#17
K M Patel, K E van der Vos, C G Smith, F Mouliere, D Tsui, J Morris, D Chandrananda, F Marass, D van den Broek, D E Neal, V J Gnanapragasam, T Forshew, B W van Rhijn, C E Massie, N Rosenfeld, M S van der Heijden
Muscle Invasive Bladder Cancer (MIBC) has a poor prognosis. Whilst patients can achieve a 6% improvement in overall survival with Neo-Adjuvant Chemotherapy (NAC), many do not respond. Body fluid mutant DNA (mutDNA) may allow non-invasive identification of treatment failure. We collected 248 liquid biopsy samples including plasma, cell pellet (UCP) and supernatant (USN) from spun urine, from 17 patients undergoing NAC. We assessed single nucleotide variants and copy number alterations in mutDNA using Tagged-Amplicon- and shallow Whole Genome- Sequencing...
July 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28713371/regulation-of-subunit-specific-germinal-center-b-cell-responses-to-the-hiv-1-envelope-glycoproteins-by-antibody-mediated-feedback
#18
Mattias N E Forsell, Linda Kvastad, Saikiran K Sedimbi, John Andersson, Mikael C I Karlsson
The regulation of germinal center (GC) B cell responses to single epitopes is well investigated. How monoclonal B cells are regulated within the polyclonal B cell response to protein antigens is less so. Here, we investigate the primary GC B cell response after injection of mice with HIV-1 envelope glycoproteins. We demonstrate that single GCs are seeded by a diverse number of B cell clones shortly after a single immunization and that the presence of Env-specific antibodies can inhibit the development of early GC B cells...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28704602/defective-early-b-cell-tolerance-checkpoints-in-sj%C3%A3-gren-s-syndrome-patients
#19
Salomé Glauzy, Joel Sng, Jason M Bannock, Jacques-Eric Gottenberg, Anne-Sophie Korganow, Patrice Cacoub, David Saadoun, Eric Meffre
OBJECTIVE: Central and peripheral B cell tolerance checkpoints are defective in many patients with autoimmune diseases, but the functionality of each discrete checkpoint has not been assessed in patients with Sjögren's syndrome (SS). We undertook this study to assess this functionality in SS patients. METHODS: Using a polymerase chain reaction-based approach that allows us to clone and express, in vitro, recombinant antibodies produced by single B cells, we tested the reactivity of recombinant antibodies cloned from single CD19+CD21(low) CD10+IgM(high) CD27- newly emigrant/transitional B cells and CD19+CD21+CD10-IgM+CD27- mature naive B cells from 5 SS patients...
July 13, 2017: Arthritis & Rheumatology
https://www.readbyqxmd.com/read/28697807/inferring-clonal-structure-in-htlv-1-infected-individuals-towards-bridging-the-gap-between-analysis-and-visualization
#20
Amir Farmanbar, Sanaz Firouzi, Wojciech Makałowski, Masako Iwanaga, Kaoru Uchimaru, Atae Utsunomiya, Toshiki Watanabe, Kenta Nakai
BACKGROUND: Human T cell leukemia virus type 1 (HTLV-1) causes adult T cell leukemia (ATL) in a proportion of infected individuals after a long latency period. Development of ATL is a multistep clonal process that can be investigated by monitoring the clonal expansion of HTLV-1-infected cells by isolation of provirus integration sites. The clonal composition (size, number, and combinations of clones) during the latency period in a given infected individual has not been clearly elucidated...
July 11, 2017: Human Genomics
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