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Journal of Immunological Methods

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https://www.readbyqxmd.com/read/28652011/development-and-evaluation-of-polyclonal-antisera-for-detection-of-the-igm-heavy-chain-of-multiple-fish-species
#1
Walissara Jirapongpairoj, Ikuo Hirono, Hidehiro Kondo
Antibodies are widely considered to be essential tools for detection of immune responses in various fish species. Here we produced the peptide polyclonal antisera (anti-fish IgH-1 and anti-fish IgH-2) to detect IgM of various fish species. The peptides were designed based on the conserved sequence of the fish immunoglobulin heavy chains of seven fish species (Japanese flounder, seabream, yellowtail, carp, rainbow trout, hybrid sturgeon and banded houndshark). By Western blotting, anti-fish IgH-1 antiserum detected the IgMs of all fish species except banded houndshark...
June 23, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28648385/fluorescence-imaging-of-lymphatic-outflow-of-cerebrospinal-fluid-in-mice
#2
Sunkuk Kwon, Christopher F Janssen, Fred Christian Velasquez, Eva M Sevick-Muraca
Cerebrospinal fluid (CSF) is known to be reabsorbed by the lymphatic vessels and drain into the lymph nodes (LNs) through peripheral lymphatic vessels. In the peripheral lymphatics, the contractile pumping action of lymphangions mediates lymph drainage; yet it is unknown whether lymphatic vessels draining cranial and spinal CSF show similar function. Herein, we used non-invasive near-infrared fluorescence imaging (NIRFI) to image (i) indocyanine green (ICG) distribution along the neuraxis and (ii) routes of ICG-laden CSF outflow into the lymphatics following intrathecal lumbar administration...
June 22, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28648384/a-novel-and-rapid-method-to-quantify-treg-mediated-suppression-of-cd4-t-cells
#3
Anna E Long, Megan Tatum, Carmen Mikacenic, Jane H Buckner
Measuring regulatory T cell suppression provides important insight into T cell dysfunction in autoimmune disease. However, to date, suppression assays are limited by the requirement for freshly isolated cells, and significant cell numbers. Here, we present a novel and rapid in vitro assay using effector T cell surface expression of both CD25 and CD134 as a surrogate marker of regulatory T cell-mediated suppression. This surface marker-based suppression assay works for frozen samples and for samples with limited cell numbers...
June 22, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28647457/quantitative-in-vitro-and-in-vivo-models-to-assess-human-ige-b-cell-receptor-crosslinking-by-ige-and-empd-ige-targeting-antibodies
#4
Benjamin Vigl, Nina Salhat, Michela Parth, Halyna Pankevych, Andreas Mairhofer, Stefan Bartl, Oskar W Smrzka
Targeting plasma IgE by therapeutic mABs like Omalizumab (Xolair®) is current clinical practice for severe allergic conditions or other IgE related diseases like chronic urticaria. As an alternative to soluble IgE targeting, IgE supply can be lowered by targeting the Extracellular Membrane Proximal Domain (EMPD) of the IgE B cell receptor (BCR) present on IgE switched B cells. This ultimately leads to apoptosis of these cells upon IgE BCR crosslinking. Since tools to selectively assess the efficacy of IgE BCR crosslinking by IgE targeting antibodies are limited, a readily quantifiable cell model was developed that allows to specifically address IgE BCR crosslinking activity in vitro...
June 21, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28647456/improved-recovery-of-functionally-active-eosinophils-and-neutrophils-using-novel-immunomagnetic-technology
#5
Kiho Son, Manali Mukherjee, Brendan A S McIntyre, Jose C Eguez, Katherine Radford, Nicola LaVigne, Caroline Ethier, Francis Davoine, Luke Janssen, Paige Lacy, Parameswaran Nair
Clinically relevant and reliable reports derived from in vitro research are dependent on the choice of cell isolation protocols adopted between different laboratories. Peripheral blood eosinophils are conventionally isolated using density-gradient centrifugation followed by immunomagnetic selection (positive/negative) while neutrophils follow a more simplified dextran-sedimentation methodology. With the increasing sophistication of molecular techniques, methods are now available that promise protocols with reduced user-manipulations, improved efficiency, and better yield without compromising the purity of enriched cell populations...
June 21, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28647455/comparison-of-mucosal-lining-fluid-sampling-methods-and-influenza-specific-iga-detection-assays-for-use-in-human-studies-of-influenza-immunity
#6
Thushan I de Silva, Victoria Gould, Nuredin I Mohammed, Alethea Cope, Adam Meijer, Ilse Zutt, Johan Reimerink, Beate Kampmann, Katja Hoschler, Maria Zambon, John S Tregoning
We need greater understanding of the mechanisms underlying protection against influenza virus to develop more effective vaccines. To do this, we need better, more reproducible methods of sampling the nasal mucosa. The aim of the current study was to compare levels of influenza virus A subtype-specific IgA collected using three different methods of nasal sampling. Samples were collected from healthy adult volunteers before and after LAIV immunization by nasal wash, flocked swabs and Synthetic Absorptive Matrix (SAM) strips...
June 21, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28645528/long-term-sustainable-dendritic-cell-specific-depletion-murine-model-for-periodontitis-research
#7
Amanda Finger Stadler, Mitulkumar Patel, Rafal Pacholczyk, Christopher W Cutler, Roger M Arce
Dendritic cells (DCs) are specialized antigen-presenting cells that play a pivotal role in the pathogenesis of periodontitis. The use of animal models to study the role of DCs in periodontitis has been limited by lack of a method for sustained depletion of DCs. Hence, the objectives of this study were to validate the zDC-DTR knockin mouse model of conventional DCs (cDCs) depletion, as well as to investigate whether this depletion could be sustained long enough to induce alveolar bone loss in this model. zDC-DTR mice were treated with different dose regimens of diphtheria toxin (DT) to determine survival rate...
June 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28629733/an-optimized-protocol-for-the-generation-and-functional-analysis-of-human-mast-cells-from-cd34-enriched-cell-populations
#8
Yuzhi Yin, Yun Bai, Ana Olivera, Avanti Desai, Dean D Metcalfe
The culture of mast cells from human tissues such a cord blood, peripheral blood or bone marrow aspirates has advanced our understanding of human mast cells (huMC) degranulation, mediator production and response to pharmacologic agents. However, existing methods for huMC culture tend to be laborious and expensive. Combining technical approaches from several of these protocols, we designed a simplified and more cost effective approach to the culture of mast cells from human cell populations including peripheral blood and cryopreserved cells from lymphocytapheresis...
June 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28629732/impact-of-granulocyte-contamination-on-pbmc-integrity-of-shipped-blood-samples-implications-for-multi-center-studies-monitoring-regulatory-t-cells
#9
Charuta Agashe, David Chiang, Alexander Grishin, Madhan Masilamani, Stacie M Jones, Robert A Wood, Scott H Sicherer, A Wesley Burks, Donald Y M Leung, Peter Dawson, Hugh A Sampson, M Cecilia Berin
In centralized immune monitoring for a multi-center allergen immunotherapy trial, we observed frequent loss of CD4(+) T cell integrity following staining of cultured PBMCs with our regulatory T cell flow cytometry panel. Samples were marked by a loss of total cellular events, altered scatter properties, and reduced CD3(+)CD4(+) events. This occurred only in samples that were stained with Foxp3 and were therefore treated with Foxp3 fixation-permeabilization buffer. We identified granulocyte contamination in samples associated with a loss of integrity, and went on to test the impact of granulocyte depletion on day-old blood samples...
June 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28625864/a-systematic-study-of-the-effect-of-low-ph-acid-treatment-on-anti-drug-antibodies-specific-for-a-domain-antibody-therapeutic-impact-on-drug-tolerance-assay-sensitivity-and-post-validation-method-assessment-of-ada-in-clinical-serum-samples
#10
Uma Kavita, Jia Duo, Sean M Crawford, Rong Liu, Joan Valcin, Carol Gleason, Huijin Dong, Snaehal Gadkari, Robert W Dodge, Renuka C Pillutla, Binodh S DeSilva
We developed a homogeneous bridging anti-drug antibody (ADA) assay on an electro chemiluminescent immunoassay (ECLIA) platform to support the immunogenicity evaluation of a dimeric domain antibody (dAb) therapeutic in clinical studies. During method development we evaluated the impact of different types of acid at various pH levels on polyclonal and monoclonal ADA controls of differing affinities and on/off rates. The data shows for the first time that acids of different pH can have a differential effect on ADA of various affinities and this in turn impacts assay sensitivity and drug tolerance as defined by these surrogate controls...
June 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28614699/a-signal-amplification-probe-enhances-sensitivity-of-antibodies-and-aptamers-based-immuno-diagnostic-assays
#11
Ofer Nussbaum, Michal Bar Oz, Tal Tilayov, Helly Atiya, Shlomo Dagan
One major unmet need is improving the sensitivity of immune-diagnostic assays. This is particularly important in the field of biomarker discoveries and monitoring. We have established a novel signal amplification probe system enabling a highly sensitive target detection platform to be used in immuno-assays. The probe consists of a double stranded DNA that can carry a large number of signaling elements such as biotin or fluorescent molecules. The DNA probe anchors to the recognition unit, whether an antibody or an aptamer, by covalent conjugation or by a simple and rapid molecular association process...
June 11, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28579366/-two-in-one-approach-for-bioassay-selection-for-dual-specificity-antibodies
#12
Ho Young Lee, Gabriele Schaefer, Chingwei Vivian Lee, Pin Yee Wong, Guoying Jiang
Dual specific antibodies and bispecific antibodies that recognize two different antigen targets are currently being regarded as very effective therapeutics for complex human diseases. While effective, designing and developing a bioassay strategy for dual specific antibodies that is reflective of the mechanism of action (MoA) and also measures the dual activities of antibodies pose unique and exciting challenges. An important question asked while developing a bioassay for dual specific antibodies is, "How many bioassays will be needed, one bioassay or two separate bioassays?" Here we present an approach of using one bioassay for a dual specific antibody that targets two receptors in signaling pathways...
June 1, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28579365/a-novel-python-program-for-implementation-of-quality-control-in-elisa
#13
Hanna N Wetzel, Cinder Cohen, Andrew B Norman, Rose P Webster
The use of semi-quantitative assays such as the enzyme-linked immunosorbent assay (ELISA) requires stringent quality control of the data. However, such quality control is often lacking in academic settings due to unavailability of software and knowledge. Therefore, our aim was to develop methods to easily implement Levey-Jennings quality control methods. For this purpose, we created a program written in Python (a programming language with an open source license) and tested it using a training set of ELISA standard curves quantifying the Fab fragment of an anti-cocaine monoclonal antibody in mouse blood...
June 1, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28576653/a-g-protein-coupled-chemokine-receptor-a-putative-insertion-site-for-a-multi-pathogen-recombinant-capripoxvirus-vaccine-strategy
#14
Catherine Cêtre-Sossah, Simon Dickmu, Olivier Kwiatek, Emmanuel Albina
Capripoxviruses (CaPVs) have been shown to be ideal viral vectors for the development of recombinant multivalent vaccines to enable delivery of immunogenic genes from ruminant pathogens. So far, the viral thymidine kinase (TK) gene is the only gene used to generate recombinants. A putative non-essential gene encoding a G-protein-coupled chemokine receptor subfamily homologue (GPCR) was targeted as an additional insertion site. Peste des petits ruminants (PPR) were chosen as a disease model. A new recombinant CaPV expressing the viral attachment hemagglutinin (H) of the PPR virus (PPRV) in the GPCR insertion site (rKS1-HPPR-GPCR) was generated in the backbone North African isolate KS1 strain of lumpy skin disease virus (LSDV)...
May 30, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28554544/immuno-nephelometric-determination-of-group-streptococcal-anti-streptolysin-o-titres-asot-from-dried-blood-spots-method-for-validating-a-new-assay
#15
John Joseph, Neil Kent, Asha Bowen, Julie Hart, Meru Sheel, Robert Wardrop, Sam Abbs, Scott Bazely, Monika Rybak
This study was designed to determine the sensitivity and reproducibility of recovering anti-streptolysin O titres (ASOT) from dried blood spot (DBS) samples, a methodologic subcomponent of the penicillin pharmacokinetic studies in children receiving secondary prophylaxis with intramuscular benzathine penicillin for acute rheumatic fever.
May 26, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28554543/a-method-for-the-isolation-and-characterization-of-functional-murine-monoclonal-antibodies-by-single-b-cell-cloning
#16
Sara Carbonetti, Brian G Oliver, Vladimir Vigdorovich, Nicholas Dambrauskas, Brandon Sack, Emilee Bergl, Stefan H I Kappe, D Noah Sather
Monoclonal antibody technologies have enabled dramatic advances in immunology, the study of infectious disease, and modern medicine over the past 40years. However, many monoclonal antibody discovery procedures are labor- and time-intensive, low efficiency, and expensive. Here we describe an optimized mAb discovery platform for the rapid and efficient isolation, cloning and characterization of monoclonal antibodies in murine systems. In this platform, antigen-binding splenic B cells from immunized mice are isolated by FACS and cocultured with CD40L positive cells to induce proliferation and mAb production...
May 26, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28551309/ontak-like-human-il-2-fusion-toxin
#17
Zhaohui Wang, Qian Zheng, Huiping Zhang, Roderick T Bronson, Joren C Madsen, David H Sachs, Christene A Huang, Zhirui Wang
Ontak® is a FDA-approved diphtheria toxin-based recombinant fusion toxin for treatment of human CD25(+) cutaneous T cell lymphoma (CTCL). However, it has been discontinued clinically due to the production issue related to the bacterial expression system with difficult purification. Recently we have developed monovalent and bivalent human IL-2 fusion toxins targeting human CD25(+) cells using advanced unique diphtheria toxin resistant yeast Pichia Pastoris expression system. In vitro efficacy characterization using human CD25(+) HUT102/6TG cells demonstrated that both monovalent and bivalent isoforms are potent and the bivalent isoform is approximately two logs more potent than the monovalent isoform...
May 24, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28529048/laboratory-based-performance-evaluation-of-pima-cd4-t-lymphocyte-count-point-of-care-by-lay-counselors-in-kenya
#18
Clement Zeh, Charles E Rose, Seth Inzaule, Mitesh A Desai, Fredrick Otieno, Felix Humwa, Benta Akoth, Paul Omolo, Robert T Chen, Yenew Kebede, Taraz Samandari
BACKGROUND: CD4+ T-lymphocyte count testing at the point-of-care (POC) may improve linkage to care of persons diagnosed with HIV-1 infection, but the accuracy of POC devices when operated by lay-counselors in the era of task-shifting is unknown. We examined the accuracy of Alere's Pima™ POC device on both capillary and venous blood when performed by lay-counselors and laboratory technicians. METHODS: In Phase I, we compared the perfomance of POC against FACSCalibur™ for 280 venous specimens by laboratory technicians...
May 18, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28527901/photonic-ring-resonance-is-a-versatile-platform-for-performing-multiplex-immunoassays-in-real-time
#19
Sasi Mudumba, Sophia de Alba, Randy Romero, Carli Cherwien, Alice Wu, Jue Wang, Martin A Gleeson, Muzammil Iqbal, Rufus W Burlingame
Photonic ring resonance is a property of light where in certain circumstances specific wavelengths are trapped in a ring resonator. Sensors based on silicon photonic ring resonators function by detecting the interaction between light circulating inside the sensor and matter deposited on the sensor surface. Binding of biological material results in a localized change in refractive index on the sensor surface, which affects the circulating optical field extending beyond the sensor boundary. That is, the resonant wavelength will change when the refractive index of the medium around the ring resonator changes...
May 17, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28522403/multiplex-autoantibody-detection-for-autoimmune-liver-diseases-and-autoimmune-gastritis
#20
Joris Vanderlocht, Mart van der Cruys, Frans Stals, Liesbeth Bakker-Jonges, Jan Damoiseaux
Autoantibody detection for autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and autoimmune gastritis (AIG) is traditionally performed by IIF on a combination of tissues. Multiplex line/dot blots (LIA/DIA) offer multiple advantages, i.e. automation, objective reading, no interfering reactivities, no coincidental findings. In the current study we evaluated automated DIA (D-Tek) for detecting autoantibodies related to autoimmune diseases of the gastrointestinal tract. We tested samples of the Dutch EQC program and compared the results with the consensus of the participating labs...
May 16, 2017: Journal of Immunological Methods
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