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single b cell cloning

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https://www.readbyqxmd.com/read/27906032/infection-of-rhesus-macaques-with-a-pool-of-simian-immunodeficiency-virus-with-the-envelope-genes-from-acute-hiv-1-infections
#1
Kendall C Krebs, Meijuan Tian, Mohammed Asmal, Binhua Ling, Kenneth Nelson, Kenneth Henry, Richard Gibson, Yuejin Li, Weining Han, Robin J Shattock, Ronald S Veazey, Norman Letvin, Eric J Arts, Yong Gao
BACKGROUND: New simian-human immunodeficiency chimeric viruses with an HIV-1 env (SHIVenv) are critical for studies on HIV pathogenesis, vaccine development, and microbicide testing. Macaques are typically exposed to single CCR5-using SHIVenv which in most instances does not reflect the conditions during acute/early HIV infection (AHI) in humans. Instead of individual and serial testing new SHIV constructs, a pool of SHIVenv_B derived from 16 acute HIV-1 infections were constructed using a novel yeast-based SHIV cloning approach and then used to infect macaques...
November 25, 2016: AIDS Research and Therapy
https://www.readbyqxmd.com/read/27888111/neutralizing-human-recombinant-antibodies-against-herpes-simplex-virus-type-1-glycoproteins-b-from-a-phage-displayed-scfv-antibody-library
#2
Vahid Bagheri, Foroogh Nejatollahi, Seyed Alireza Esmaeili, Amir Abbas Momtazi, Mohamad Motamedifar, Amirhossein Sahebkar
The HSV-1 envelope glycoprotein B (gB) plays a critical role in virus entry into host cells. Neutralizing antibodies can therefore potentially prevent virus entry into target cells and cell-to-cell spread of infection. Our present study focused on the selection of neutralizing single-chain Fv (scFv) antibodies of a phage-displayed nonimmune human scFv antibody library against gB of HSV-1. To enrich specific scFvs, two phage antibodies were isolated against amino acid residues 31-43 derived from the N-terminal part of gB using panning technique...
November 22, 2016: Life Sciences
https://www.readbyqxmd.com/read/27830060/clonal-selection-versus-clonal-cooperation-the-integrated-perception-of-immune-objects
#3
Serge Nataf
Analogies between the immune and nervous systems were first envisioned by the immunologist Niels Jerne who introduced the concepts of antigen "recognition" and immune "memory". However, since then, it appears that only the cognitive immunology paradigm proposed by Irun Cohen, attempted to further theorize the immune system functions through the prism of neurosciences. The present paper is aimed at revisiting this analogy-based reasoning. In particular, a parallel is drawn between the brain pathways of visual perception and the processes allowing the global perception of an "immune object"...
2016: F1000Research
https://www.readbyqxmd.com/read/27814608/lysosomal-alpha-mannosidase-and-alpha-mannosidosis
#4
Silvia Paciotti, Michela Codini, Anna Tasegian, Maria Rachele Ceccarini, Samuela Cataldi, Cataldo Arcuri, Bernard Fioretti, Elisabetta Albi, Tommaso Beccari
Lysosomal alpha-mannosidase with acidic pH optimum is ubiquitous in human tissues where is expressed in two major forms, A and B that are the product of a single gene located on chromosome 19. Mutations in the gene encoding for alpha-mannosidase cause alpha- mannosidosis, an autosomal recessive disease, resulting in the accumulation of unprocessed mannose containing oligosaccharide material. This rare disease has an estimated incidence of 1/500.0.00 live births and clinically is divided into three subgroups...
January 1, 2017: Frontiers in Bioscience (Landmark Edition)
https://www.readbyqxmd.com/read/27811010/dual-inhibition-of-mek-and-pi3k-akt-rescues-cancer-cachexia-through-both-tumor-extrinsic-and-intrinsic-activities
#5
Erin E Talbert, Jennifer Yang, Thomas A Mace, Matthew R Farren, Alton B Farris, Gregory S Young, Omar Elnaggar, Zheng Che, Cynthia D Timmers, Priyani Rajasekera, Jennifer M Maskarinec, Mark Bloomston, Tanios Bekaii-Saab, Denis C Guttridge, Gregory B Lesinski
Involuntary weight loss, a part of the cachexia syndrome, is a debilitating co-morbidity of cancer and currently has no treatment options. Results from a recent clinical trial at our institution showed that biliary tract cancer patients treated with a MEK inhibitor exhibited poor tumor responses, but surprisingly gained weight and increased their skeletal muscle mass. This implied that MEK inhibition might be anti-cachectic. To test this potential effect of MEK inhibition, we utilized the established Colon-26 model of cancer cachexia and the MEK½ inhibitor MEK162...
November 3, 2016: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/27798614/analysis-of-self-antigen-specificity-of-islet-infiltrating-t-cells-from-human-donors-with-type-1-diabetes
#6
Jenny Aurielle B Babon, Megan E DeNicola, David M Blodgett, Inne Crèvecoeur, Thomas S Buttrick, René Maehr, Rita Bottino, Ali Naji, John Kaddis, Wassim Elyaman, Eddie A James, Rachana Haliyur, Marcela Brissova, Lut Overbergh, Chantal Mathieu, Thomas Delong, Kathryn Haskins, Alberto Pugliese, Martha Campbell-Thompson, Clayton Mathews, Mark A Atkinson, Alvin C Powers, David M Harlan, Sally C Kent
A major therapeutic goal for type 1 diabetes (T1D) is to induce autoantigen-specific tolerance of T cells. This could suppress autoimmunity in those at risk for the development of T1D, as well as in those with established disease who receive islet replacement or regeneration therapy. Because functional studies of human autoreactive T cell responses have been limited largely to peripheral blood-derived T cells, it is unclear how representative the peripheral T cell repertoire is of T cells infiltrating the islets...
December 2016: Nature Medicine
https://www.readbyqxmd.com/read/27779190/single-cell-lineage-tracing-in-the-mammary-gland-reveals-stochastic-clonal-dispersion-of-stem-progenitor-cell-progeny
#7
Felicity M Davis, Bethan Lloyd-Lewis, Olivia B Harris, Sarah Kozar, Douglas J Winton, Leila Muresan, Christine J Watson
The mammary gland undergoes cycles of growth and regeneration throughout reproductive life, a process that requires mammary stem cells (MaSCs). Whilst recent genetic fate-mapping studies using lineage-specific promoters have provided valuable insights into the mammary epithelial hierarchy, the true differentiation potential of adult MaSCs remains unclear. To address this, herein we utilize a stochastic genetic-labelling strategy to indelibly mark a single cell and its progeny in situ, combined with tissue clearing and 3D imaging...
October 25, 2016: Nature Communications
https://www.readbyqxmd.com/read/27737507/oncogenic-events-rather-than-antigen-selection-pressure-may-be-the-main-driving-forces-for-relapse-in-diffuse-large-b-cell-lymphomas
#8
David Rizzo, Pierre-Julien Viailly, Sylvain Mareschal, Elodie Bohers, Jean-Michel Picquenot, Dominique Penther, Sydney Dubois, Vinciane Marchand, Philippe Bertrand, Catherine Maingonnat, Pascaline Etancelin, Jean Feuillard, Christian Bastard, Hervé Tilly, Fabrice Jardin, Philippe Ruminy
Little is known on the phylogenetic relationship between diagnostic and relapse clones of Diffuse Large B-Cell Lymphoma (DLBCL). We applied High Throughput Sequencing (HTS) of the VDJ locus of Immunoglobulin Heavy chain (IGHV) on fourteen DLBCL patients with serial samples, including tumor biopsies and/or peripheral blood mononuclear cells (PBMC). Phylogenetic data were consolidated with targeted sequencing and cytogenetics. Phylogeny clearly showed that DLBCL relapse could occur according either an early or a late divergent mode...
October 13, 2016: American Journal of Hematology
https://www.readbyqxmd.com/read/27733338/construction-of-ctcf-degradation-cell-line-by-crispr-cas9-mediated-genome-editing
#9
Xie Dejian, Shi Minglei, Zhang Yan, Wang Tianyi, Shen Wenlong, Ye Bingyu, Li Ping, He Chao, Zhang Xiangyuan, Zhao Zhihu
The CCCTC-binding factor (CTCF) is the main insulator protein described in vertebrates. It plays fundamental roles during diverse cellular processes. CTCF gene knockout mice led to death during embryonic development. To further explore the functions of CTCF, we employed a CRISPR/Cas9-based genome engineering strategy to in-frame insert the mitosis-special degradation domain (MD) of cyclin B into the upstream open reading frame of CTCF gene. Fusion protein is designed to degrade during mitosis leaded by MD. As a control group, mutation of a single arginine (R42A) within the destruction box inactivates the MD leading to constitutive expression of MD(*)-CTCF...
July 20, 2016: Yi Chuan, Hereditas
https://www.readbyqxmd.com/read/27708334/loss-of-immune-tolerance-to-il-2-in-type-1-diabetes
#10
Louis Pérol, John M Lindner, Pamela Caudana, Nicolas Gonzalo Nunez, Audrey Baeyens, Andrea Valle, Christine Sedlik, Delphine Loirat, Olivier Boyer, Alain Créange, José Laurent Cohen, Ute Christine Rogner, Jun Yamanouchi, Martine Marchant, Xavier Charles Leber, Meike Scharenberg, Marie-Claude Gagnerault, Roberto Mallone, Manuela Battaglia, Pere Santamaria, Agnès Hartemann, Elisabetta Traggiai, Eliane Piaggio
Type 1 diabetes (T1D) is characterized by a chronic, progressive autoimmune attack against pancreas-specific antigens, effecting the destruction of insulin-producing β-cells. Here we show interleukin-2 (IL-2) is a non-pancreatic autoimmune target in T1D. Anti-IL-2 autoantibodies, as well as T cells specific for a single orthologous epitope of IL-2, are present in the peripheral blood of non-obese diabetic (NOD) mice and patients with T1D. In NOD mice, the generation of anti-IL-2 autoantibodies is genetically determined and their titre increases with age and disease onset...
October 6, 2016: Nature Communications
https://www.readbyqxmd.com/read/27702818/paired-high-content-analysis-of-prostate-cancer-cells-in-bone-marrow-and-blood-characterizes-increased-androgen-receptor-expression-in-tumor-cell-clusters
#11
Peter Kuhn, Anders Carlsson, Madelyn S Luttgen, Kevin Keomanee Dizon, Patricia Troncoso, Paul Corn, Anand Kolatkar, James B Hicks, Christopher Logothetis, Amado J Zurita
PURPOSE: Recent studies demonstrate that prostate cancer clones from different metastatic sites are dynamically represented in the blood of patients over time, suggesting that the paired evaluation of tumor cells in circulation and bone marrow, the primary target for prostate cancer metastasis, may provide complementary information. EXPERIMENTAL DESIGN: We adapted our single-cell high-content liquid biopsy platform to bone marrow aspirates (BMA), to concurrently identify and characterize prostate cancer cells in patients' blood and bone and thus discern features associated to tumorigenicity and dynamics of metastatic progression...
October 4, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/27698429/caspr2-reactive-antibody-cloned-from-a-mother-of-an-asd-child-mediates-an-asd-like-phenotype-in-mice
#12
L Brimberg, S Mader, V Jeganathan, R Berlin, T R Coleman, P K Gregersen, P T Huerta, B T Volpe, B Diamond
Autism spectrum disorder (ASD) occurs in 1 in 68 births, preferentially affecting males. It encompasses a group of neurodevelopmental abnormalities characterized by impaired social interaction and communication, stereotypic behaviors and motor dysfunction. Although recent advances implicate maternal brain-reactive antibodies in a causative role in ASD, a definitive assessment of their pathogenic potential requires cloning of such antibodies. Here, we describe the isolation and characterization of monoclonal brain-reactive antibodies from blood of women with brain-reactive serology and a child with ASD...
December 2016: Molecular Psychiatry
https://www.readbyqxmd.com/read/27693386/dynamic-variation-of-cd5-surface-expression-levels-within-individual-chronic-lymphocytic-leukaemia-clones
#13
Rachael J M Bashford-Rogers, Anne L Palser, Clare Hodkinson, Joanna Baxter, George A Follows, George S Vassiliou, Paul Kellam
Chronic lymphocytic leukaemia (CLL) is characterised by the accumulation of clonally-derived mature CD5(high) B-cells, however the cellular origin of CLL is still unknown. Patients with CLL also harbour variable numbers of CD5(low) B-cells, but the clonal relationship of these cells to the bulk disease is unknown and can have important implications for monitoring, treating and understanding the biology of CLL. Here we use B-cell receptors (BCRs) as molecular barcodes to first show that the great majority of CD5(low) B-cells in the blood of CLL patients are clonally related to CD5(high) CLL B-cells by single-cell BCR sequencing...
September 27, 2016: Experimental Hematology
https://www.readbyqxmd.com/read/27688876/emergence-of-linezolid-and-vancomycin-resistant-enterococcus-faecium-in-a-department-for-hematologic-stem-cell-transplantation
#14
M Krull, I Klare, B Ross, R Trenschel, D W Beelen, D Todt, E Steinmann, J Buer, P-M Rath, J Steinmann
BACKGROUND: Prevalence of vancomycin-resistant enterococci has increased in Germany. Here, we report the cluster of linezolid- and vancomycin-resistant Enterococcus faecium (LVRE) in a German department for hematologic stem cell transplantation (HSCT). METHODS: In this retrospective analysis we included all patients with LVRE in a university-based department for HSCT in 2014 and 2015. Patients chart reviews were used to investigate the epidemiology and clinical outcome...
2016: Antimicrobial Resistance and Infection Control
https://www.readbyqxmd.com/read/27658758/antibodies-and-superantibodies-in-chronic-rhinosinusitis-with-nasal-polyps
#15
Jiun-Bo Chen, Louisa K James, Anna M Davies, Y-C Wu, Joanne Rimmer, Valerie J Lund, Jou-Han Chen, James M McDonnell, Yih-Chih Chan, George H Hutchins, Tse Wen Chang, Brian J Sutton, Harsha H Kariyawasam, Hannah J Gould
BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with local immunoglobulin hyperproduction and the presence of IgE antibodies against Staphylococcus aureus enterotoxins (SAEs). Aspirin-exacerbated respiratory disease (AERD) is a severe form of CRSwNP in which nearly all patients express anti-SAEs. OBJECTIVES: We aimed to understand antibodies reactive to SAEs and determine whether they recognize SAEs through their complementarity-determining regions (CDRs) or framework regions (FRs)...
September 19, 2016: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/27649554/the-t-cell-receptor-repertoire-influences-the-tumor-microenvironment-and-is-associated-with-survival-in-aggressive-b-cell-lymphoma
#16
Colm Keane, Clare Gould, Kimberley Jones, David Hamm, Dipti Talaulikar, Jonathan Ellis, Frank Vari, Simone Birch, Erica Han, Peter Wood, Kim-Anh Le Cao, Michael R Green, Pauline Crooks, Sanjiv Jain, Joshua Tobin, Raymond J Steptoe, Maher K Gandhi
PURPOSE: To investigate the relationship between the intra-tumoral T-cell receptor (TCR) repertoire and the tumor microenvironment (TME) in de novo Diffuse Large B-cell Lymphoma (DLBCL), and the TCR's impact on survival. EXPERIMENTAL DESIGN: We performed high-throughput unbiased TCRBeta sequencing on a population based cohort of 92 DLBCL patients treated with conventional (i.e. non-checkpoint blockade) frontline 'R-CHOP' therapy. Key immune checkpoint genes within the TME were digitally quantified by nanoString{trade mark, serif}...
September 20, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/27649318/lamivudine-adefovir-treatment-increases-the-rate-of-spontaneous-mutation-of-hepatitis-b-virus-in-patients
#17
Marianoel Pereira-Gómez, Juan-Vicente Bou, Iván Andreu, Rafael Sanjuán
The high levels of genetic diversity shown by hepatitis B virus (HBV) are commonly attributed to the low fidelity of its polymerase. However, the rate of spontaneous mutation of human HBV in vivo is currently unknown. Here, based on the evolutionary principle that the population frequency of lethal mutations equals the rate at which they are produced, we have estimated the mutation rate of HBV in vivo by scoring premature stop codons in 621 publicly available, full-length, molecular clone sequences derived from patients...
2016: PloS One
https://www.readbyqxmd.com/read/27639717/selection-and-characterization-of-single-domain-antibodies-against-human-cd20
#18
Jinny L Liu, Dan Zabetakis, Ellen R Goldman, George P Anderson
CD20 is a membrane protein with four integral membrane regions and a large extracellular loop between residues 142 and 187, which serves as a target binding region for rituximab (RTX) and most other anti-CD20 monoclonal antibodies. It is highly expressed in B-lymphoma cells and B lymphocytes and often serves as a target for immunotherapy. In this study, we developed single domain antibodies (sdAbs) for the sensitive detection of CD20. To achieve this, an immune sdAb library was prepared from llamas immunized with a fusion between the large loop from CD20 and Hoc, a highly antigenic protein from the T4 bacteriophage, (CD20-Hoc)...
October 2016: Molecular Immunology
https://www.readbyqxmd.com/read/27621933/cloning-expression-and-purification-of-pseudomonas-aeruginosa-flagellin-and-characterization-of-the-elicited-anti-flagellin-antibody
#19
Bahador Behrouz, Nour Amirmozafari, Nima Khoramabadi, Mahboobeh Bahroudi, Parisa Legaee, Mehdi Mahdavi
BACKGROUND: Pseudomonas aeruginosa is an important opportunistic human pathogen that causes serious infections in immunocompromised hosts. The single polar flagellum is an important factor in both virulence and colonization. OBJECTIVES: As flagellin is the major component of the flagellar filament, the main aims of the present study are to identify, clone, express, and purify the recombinant type B flagellin (r-B-flagellin) of P. aeruginosa, as well as to evaluate the functional activity of the rabbit polyclonal antibody raised against this r-B-flagellin...
June 2016: Iranian Red Crescent Medical Journal
https://www.readbyqxmd.com/read/27600311/refined-protocol-for-generating-monoclonal-antibodies-from-single-human-and-murine-b-cells
#20
Irvin Y Ho, Jeffrey J Bunker, Steven A Erickson, Karlynn E Neu, Min Huang, Mario Cortese, Bali Pulendran, Patrick C Wilson
Generating monoclonal antibodies from single B cells is a valuable tool for characterizing the specificity and functional properties of humoral responses. We and others developed protocols that have facilitated major advances in our understanding of B cell development, tolerance, and effector responses to HIV and influenza. Here, we demonstrate various refinements and dramatically reduce the time required to produce recombinant antibodies. Further, we present new methods for cloning and isolating antibodies from cells with lower immunoglobulin mRNA levels that may be resistant to traditional techniques...
November 2016: Journal of Immunological Methods
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