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Journal of Immunological Methods

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https://www.readbyqxmd.com/read/29772250/development-and-validation-of-a-novel-semi-homogenous-clinical-assay-for-quantitation-of-ranibizumab-in-human-serum
#1
John Lowe, Eric Wakshull, Theresa Shek, Anan Chuntharapai, Rebecca Elliott, Jeriza Rusit, Mauricio Maia
Ranibizumab (Lucentis®), a humanized antigen-binding fragment (Fab) monoclonal antibody that blocks VEGF-A activity is currently approved for the treatment of several retinal degenerative diseases. The assessment of drug pharmacokinetics (PK) is essential for evaluating exposure as it relates to drug safety and efficacy. For drugs administered intravitreally, systemic drug levels during the course of clinical studies are typically 100 to 1000-fold lower than those of similar therapeutics dosed intravenously, posing a significant bioanalytical challenge for PK measurements...
May 14, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29772249/evaluation-of-the-effect-of-storage-condition-on-cell-extraction-and-flow-cytometric-analysis-from-intestinal-biopsies
#2
Manon E Wildenberg, Marjolijn Duijvestein, Liset Westera, Tanja van Viegen, Christianne J Buskens, Jarmila D W van der Bilt, Larry Stitt, Vipul Jairath, Brian G Feagan, Niels Vande Casteele
BACKGROUND: Flow cytometric (FC) analysis of intestinal tissue biopsies requires prompt cell isolation and processing to prevent cell death and generate valid data. We examined the effect of storage conditions prior to cell isolation and FC on viable cell yield and the proportions of immune cell phenotypes from intestinal biopsies. METHODS: Biopsies (N = 224) from inflamed or non-inflamed ileal and/or colonic tissue from three patients with Crohn's disease were processed and analyzed immediately in duplicate, or stored under different conditions...
May 14, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29758224/humanised-recombinant-antibody-fragments-bind-human-pancreatic-islet-cells
#3
Abigail C Buenafe, Philip R Streeter
We describe here the humanisation of two mouse monoclonal antibodies that bind to surface markers on human pancreatic islet endocrine cells. Monoclonal antibodies produced by the HIC1-2B4 and HIC0-4F9 mouse hybridomas bind distinct surface molecules expressed on endocrine cells and have been validated for a number of experimental methods including immunohistochemistry and live cell sorting by flow cytometry. Variable region framework and first constant region domain sequences were replaced with that from compatible human antibody sequences, and the resulting recombinant antigen-binding fragments were cloned and expressed in mouse myeloma cells...
May 11, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29758223/letter-to-the-editor
#4
LETTER
Jar-How Lee
No abstract text is available yet for this article.
May 11, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29689263/analysis-of-glycogen-metabolic-pathway-utilization-by-dendritic-cells-and-t-cells-using-custom-phenotype-metabolic-assays
#5
Phyu Thwe, Eyal Amiel
In the field of immunology, there is an increasing interest in cellular energy metabolism and its outcome on immune cell effector function. Activation of immune cells leads to rapid metabolic changes that are central to cellular biology in order to support the effector responses. Therefore, the need for user-friendly and dependable assay technologies to address metabolic regulation and nutrient utilization in immune cells is an important need in this field. Redox-dye reduction-based Phenotype MicroArray (PM) assays, which measure NADH reduction as a readout, developed by Biolog Inc...
April 21, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29684430/development-of-an-rna-based-kit-for-easy-generation-of-tcr-engineered-lymphocytes-to-control-t-cell-assay-performance
#6
Nicole Bidmon, Sonja Kind, Marij J P Welters, Deborah Joseph-Pietras, Karoline Laske, Dominik Maurer, Sine Reker Hadrup, Gerty Schreibelt, Richard Rae, Ugur Sahin, Cécile Gouttefangeas, Cedrik M Britten, Sjoerd H van der Burg
Cell-based assays to monitor antigen-specific T-cell responses are characterized by their high complexity and should be conducted under controlled conditions to lower multiple possible sources of assay variation. However, the lack of standard reagents makes it difficult to directly compare results generated in one lab over time and across institutions. Therefore TCR-engineered reference samples (TERS) that contain a defined number of antigen-specific T cells and continuously deliver stable results are urgently needed...
April 20, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29684429/simultaneous-in-vitro-generation-of-human-cd34-derived-dendritic-cells-and-mast-cells-from-non-mobilized-peripheral-blood-mononuclear-cells
#7
Pavla Taborska, Jirina Bartunkova, Daniel Smrz
Dendritic cells (DCs) and mast cells (MCs) are key players of the immune system, often coming in close proximity in peripheral tissues. The interplay of these cells is, however, still poorly understood especially with regards to human cells. The reason for that is the absence of a well established in vitro human cell-based study system that would allow a simultaneous preparation of both cell types. In this study, we show a method for simultaneous generation of DCs and MCs from CD34+ stem cell progenitors that were isolated from the non-adherent fraction of non-mobilized peripheral blood mononuclear cells of healthy donors...
April 20, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29684428/validation-of-a-cd1b-tetramer-assay-for-studies-of-human-mycobacterial-infection-or-vaccination
#8
Erik D Layton, Krystle K Q Yu, Malisa T Smith, Thomas J Scriba, Stephen C De Rosa, Chetan Seshadri
CD1 tetramers loaded with lipid antigens facilitate the identification of rare lipid-antigen specific T cells present in human blood and tissue. Because CD1 proteins are structurally non-polymorphic, these tetramers can be applied to genetically diverse human populations, unlike MHC-I and MHC-II tetramers. However, there are no standardized assays to quantify and characterize lipid antigen-specific T cells present within clinical samples. We incorporated CD1b tetramers loaded with the mycobacterial lipid glucose monomycolate (GMM) into a multi-parameter flow cytometry assay...
April 20, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29684427/relationship-between-the-frequency-of-influenza-vaccination-and-cell-mediated-immunity
#9
Naruhito Otani, Masayuki Shima, Takashi Ueda, Kaoru Ichiki, Kazuhiko Nakajima, Yoshio Takesue, Kenta Honjo, Natsuko Yoshida, Sayo Kawata, Toshiomi Okuno
Despite established guidelines for population-level assessments of immunity after vaccination, standard methods for individual-level analyses have not been established, limiting the ability to optimize vaccination strategies within a particular season. In this study, we evaluated changes in cell-mediated immunity (CMI) with respect to the number of influenza vaccine doses. In particular, the influenza vaccine was administered to 21 adults during the 2015-2016 season. IFN-γ production induced by the influenza vaccine antigens [A (H1N1), A (H3N2), B (Yamagata lineage), and B (Victoria lineage)] increased after the first dose of vaccination in 11, 10, 10, and 11 subjects, respectively...
April 20, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29654816/a-novel-method-for-sensitive-low-cost-and-portable-detection-of-hepatitis-b-surface-antigen-using-a-personal-glucose-meter
#10
Saeed Taebi, Mehrnaz Keyhanfar, Abdollah Noorbakhsh
Hepatitis B virus (HBV) infection is the major public health problem leading cause of death worldwide. The most important diagnostic marker for this infection is hepatitis B surface antigen (HBsAg). In this study, a novel, inexpensive, portable and sensitive ELISA method was designed and investigated for diagnosis of HBsAg based on the functionalized Fe3 O4 and Al2 O3 nanoparticles, with the strategy for detecting the concentration of glucose using a cheap and accessible personal glucose meter (PGM). The ELISA system was constructed using hepatitis B antibody against HBsAg immobilized on streptavidin coated magnetic iron oxide particles (S-Fe3 O4 ) as the capture antibody (Ab1 )...
April 11, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29654815/quantification-of-nk-cell-activity-using-whole-blood-methodological-aspects-of-a-new-test
#11
Line Nederby, Anders Jakobsen, Marianne Hokland, Torben Frostrup Hansen
Natural Killer (NK) cells are essential in the biological fight against cancer and intracellular pathogens, and their level of activity has in many settings been used as a biomarker for a functional immune response. Currently, NK cell activity is measured using either 51 Cr-release assays or flow cytometry based assays revealing the cells´ cytotoxic capacity or by stimulating them to produce cytokines. Although very effective, these are cumbersome techniques not suitable for high volume clinical laboratories...
April 11, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29630967/inducible-disruption-of-the-c-myb-gene-allows-allogeneic-bone-marrow-transplantation-without-irradiation
#12
C Stremmel, R Schuchert, V Schneider, T Weinberger, R Thaler, D Messerer, S Helmer, F Geissmann, J Frampton, S Massberg, C Schulz
Allogeneic bone marrow (BM) transplantation enables the in vivo functional assessment of hematopoietic cells. As pre-conditioning, ionizing radiation is commonly applied to induce BM depletion, however, it exerts adverse effects on the animal and can limit experimental outcome. Here, we provide an alternative method that harnesses conditional gene deletion to ablate c-myb and thereby deplete BM cells, hence allowing BM substitution without other pre-conditioning. The protocol results in a high level of blood chimerism after allogeneic BM transplantation, whereas immune cells in peripheral tissues such as resident macrophages are not replaced...
April 6, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29625078/automated-generation-of-immature-dendritic-cells-in-a-single-use-system
#13
Andrew Kozbial, Lekhana Bhandary, Bradley B Collier, Christopher S Eickhoff, Daniel Hoft, Shashi K Murthy
Dendritic cells (DCs) are an indispensable part of studying human responses that are important for protective immunity against cancer and infectious diseases as well as prevention of autoimmunity and transplant rejection. These cells are also key elements of personalized vaccines for cancer and infectious diseases. Despite the vital role of DCs in both clinical and basic research contexts, methods for obtaining these cells from individuals remains a comparatively under-developed and inefficient process. DCs are present in very low concentrations (<1%) in blood, thus they must be generated from monocytes and the current methodology in DC generation involves a laborious process of static culture and stimulation with cytokines contained in culture medium...
April 3, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29625077/optimising-the-collection-of-female-genital-tract-fluid-for-cytokine-analysis-in-pregnant-women
#14
C S Short, R Quinlan, P Bennett, R J Shattock, G P Taylor
INTRODUCTION: To better understand the immunology of pregnancy, study of female genital tract fluid (FGF) is desirable. However the optimum method of collection of FGF in pregnant women for immunological methods, specifically cytokine measurement, is unknown. METHODS: A prospective study of HIV-uninfected pregnant women comparing two methods of FGF collection: polyvinyl acetal sponge collection of cervical fluid (CF) and menstrual cup collection of cervicovaginal fluid (CVF)...
April 3, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29625076/lymphatic-cannulation-models-in-sheep-recent-advances-for-immunological-and-biomedical-research
#15
REVIEW
Elizabeth A Washington, Stuart R Barber, Christina M Murray, Helen M S Davies, Wayne G Kimpton, Hung-Hsun Yen
Lymphatic cannulation models are useful tools for studying the immunobiology of the lymphatic system and the immunopathology of specific tissues in diseases. Sheep cannulations have been used extensively, as models for human physiology, fetal and neonatal development, human diseases, and for studies of ruminant pathobiology. The development of new and improved cannulation techniques in recent years has meant that difficult to access sites, such as mucosal associated tissues, are now more readily available to researchers...
April 3, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29625075/a-novel-and-sensitive-functional-assay-for-complement-factor-i-based-on-the-third-proteolytic-clip-of-c3b
#16
Peter J Lachmann, Elizabeth Lay, David J Seilly
A sensitive assay for the functional activity of complement Factor I is described. This is based on its third proteolytic clip whereby Factor I cleaves cell-bound iC3b to cell-bound C3dg and soluble C3c, thereby abolishing conglutination of the cells. Factor H is required as a co-factor for Factor I activity. Because of the low affinity of iC3b for Factor H, the assay needs to be performed at low ionic strength. This assay is easier to perform than those based on the conversion of C3b to iC3b (the first two Factor I clips), there being no need for the unstable intermediate EAC142 or for purified C3...
April 3, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29614267/influence-of-delayed-sample-processing-on-blood-immune-cell-phenotypes-immune-cell-responses-and-serum-anti-influenza-vaccine-antibody-titres
#17
Vivian Castro-Herrera, Mark Lown, George Lewith, Elizabeth A Miles, Philip C Calder
Provision of blood from distant research partners to a central laboratory can result in delayed blood processing prior to assessment of immune parameters. It is important to evaluate the effect of such delays on immune parameters. This study investigated the effect of storage of blood at room temperature for up to 72 h prior to processing and analysis on a range of immune parameters. Blood was collected from 10 healthy participants and analysed immediately (day 0) or after storage at room temperature for 24, 48 or 72 h (days 1, 2 and 3)...
March 31, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29614266/genetic-engineering-in-primary-human-b-cells-with-crispr-cas9-ribonucleoproteins
#18
Chung-An M Wu, Theodore L Roth, Yuriy Baglaenko, Dario M Ferri, Patrick Brauer, Juan Carlos Zuniga-Pflucker, Kristina W Rosbe, Joan E Wither, Alexander Marson, Christopher D C Allen
Genome editing in human cells with targeted nucleases now enables diverse experimental and therapeutic genome engineering applications, but extension to primary human B cells remains limited. Here we report a method for targeted genetic engineering in primary human B cells, utilizing electroporation of CRISPR-Cas9 ribonucleoproteins (RNPs) to introduce gene knockout mutations at protein-coding loci with high efficiencies that in some cases exceeded 80%. Further, we demonstrate knock-in editing of targeted nucleotides with efficiency exceeding 10% through co-delivery of oligonucleotide templates for homology directed repair...
March 31, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29605231/neutrophils-mediate-hiv-specific-antibody-dependent-phagocytosis-and-adcc
#19
Matthew J Worley, Kuangyu Fei, Adam J Lopez-Denman, Anthony D Kelleher, Stephen J Kent, Amy W Chung
There is growing evidence to support the role of Fc-mediated effector functions, such as Antibody-Dependent Cellular cytotoxicity (ADCC) and Antibody-Dependent Phagocytosis (ADP) in the protection and control of HIV. The RV144 trial and other recent HIV vaccine studies have highlighted the importance of ADCC responses in protection against HIV. The role of neutrophils, the most abundant leukocyte in the blood, has not been thoroughly evaluated for Fc-mediated effector functions to HIV. We optimized HIV-specific neutrophil ADCC and Antibody-Dependent Neutrophil Phagocytosis (ADNP) assays using freshly isolated primary human neutrophils from blood...
March 29, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29605230/an-efficient-protocol-for-in-vivo-labeling-of-proliferating-epithelial-cells
#20
Chloé Michel, Rita Küchler, Iris Martin, Bruno Kyewski, Sheena Pinto
The study of organogenesis, tissue-homeostasis and regeneration requires the precise assessment of in vivo cell proliferation. To this end a host of methods have been developed to detect and quantify DNA synthesis in proliferating cells. These include cell labeling with various nucleotide analogues and fluorescence reporter-based animal models with each method presenting its idiosyncratic shortcomings. Quantitative assessment of epithelial cell turnover has been partly hampered due to their variable and limited in vivo accessibility and the requirement for harsher isolation procedures to procure single cells for FACS analysis...
March 29, 2018: Journal of Immunological Methods
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