Abstract：
:The in vitro assessment of T-cell-mediated cytotoxicity plays an important and increasingly relevant role both in preclinical target evaluation and during immunomonitoring to accompany clinical trials employing targeted immunotherapies. For a long time, the gold standard for this purpose has been the chromium release assay (CRA). This end point assay, however, shows several disadvantages including the inevitable use of radioactivity. Based on electrical impedance measurements (using the xCELLigence system), we have established a label-free assay, facilitating the real-time monitoring of T-cell-mediated cytotoxicity. The coculture of peptide-specific T-cell lines with peptide-loaded target cells reproducibly led to a decrease in impedance due to induced apoptosis and detachment of target cells. Comparing our results to the standard CRA assay, we could demonstrate that impedance-based measurements show comparable results after short incubation periods (6h) but outperform the CRA both in reproducibility and sensitivity after prolonged incubation (24h), enabling the detection of target cell lysis with an effector to target ratio as low as 0.05:1. The impedance-based assay represents a valuable and highly sensitive tool for label-free real-time high throughput analysis of T-cell-mediated cytotoxicity.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Peper JK,Schuster H,Löffler MW,Schmid-Horch B,Rammensee HG,Stevanović Sdoi
10.1016/j.jim.2014.01.012subject
Has Abstractpub_date
2014-03-01 00:00:00pages
192-8eissn
0022-1759issn
1872-7905pii
S0022-1759(14)00027-1journal_volume
405pub_type
杂志文章abstract：OBJECTIVES:To report on a novel immunofluorescence pattern specifically associated with antibodies to SS-A/Ro. METHODS:A novel immunofluorescence pattern, herein designated SS-A/Ro pattern, was characterized as myriad discrete fine speckles throughout the nucleus. Eighty-six sequential samples presenting the SS-A/Ro p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
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abstract：:The haem enzyme myeloperoxidase (MPO) (EC 1.11.1.7) with a spectral A430/A280 ratio greater than 0.7 and a specific activity of 125 U/mg was purified from isolated human neutrophils. To obtain a radioimmunoassay (RIA) for this enzyme, a specific antiserum against human neutrophil MPO was raised in rabbits and used at ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90023-9
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abstract：:Proper evaluation of immunogenicity during clinical development of biotherapeutics is a major challenge to bioanalytical scientists, in part due to matrix interference in anti-drug antibody (ADA) and neutralizing antibody (NAB) assays. If not addressed, matrix interference could confound the immunogenicity assessment ...
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abstract：:Although serum-free media have been used to expand lymphokine-activated killer cells, antigen-specific CD8 T cell cytotoxicity does not develop in vitro in the absence of serum. The immunodominant Vbeta17 response to an influenza A matrix protein epitope restricted by HLA A2.1 was used to study the serum requirement f...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00026-5
更新日期：1999-05-27 00:00:00
abstract：BACKGROUND:The role of CD4(+) regulatory T cells (Tregs) and their subsets during HIV infection is controversial. Cryopreserved peripheral blood mononuclear cells (PBMC) are an important source for assessing number and function of Tregs. However, it is unknown if PBMC isolation and cryopreservation affect the expressio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.02.004
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abstract：:A simple semi-automated method for the assay of large numbers of replicate microcultures for the presence of antibody-forming cell clones is described. The supernatant medium is removed from microcultures by a single sharp flick on inverting the tray. The cultured cells are mixed with 0.05 ml of a plaque-revealing mix...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90346-5
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abstract：:IgM antibodies are often of low affinity (dissociation constant (Kd) > 10(-5) M) and therefore they are usually neglected as tools in, e.g., immunoassays. Previous studies have shown that low affinity biological interactions can be studied and exploited in affinity chromatography, biosensor technology and capillary el...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00039-8
更新日期：1998-05-01 00:00:00
abstract：:Many antigens are shared between different types of human leukocytes. In an effort to obtain new lineage-specific monoclonal antibodies, particularly antibodies to dendritic cells, we attempted to tolerize newborn mice to one type of leukocyte and then immunize the adults with another. We found that T cells, either un...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00335-t
更新日期：1995-04-12 00:00:00
abstract：:A sensitive direct assay has been developed to quantitate antigen specific helper factors (ThF). The assay depends on the ability of ThF to block the binding of free antibody to immobilized antigen. It has been used to follow the course of purification of ThF produced in short-term cultures of in vitro primed helper c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90098-x
更新日期：1984-02-24 00:00:00
abstract：:A new compact and cell dense continuous culture system was developed. The system consisted of a culture bag, which had two compartments separated by a semipermeable membrane, and an external rotator. Cells were cultured in the inner compartment with medium containing serum or the necessary growth factors. The outer co...
journal_title：Journal of immunological methods
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doi：10.1016/0022-1759(89)90049-5
更新日期：1989-03-10 00:00:00
abstract：:This study examined the secretion of IL-1 alpha and IL-1 beta by THP-1 leukemia cells following activation with mezerein and promotion of synthesis by interferon (IFN-gamma). Interleukin-1 (IL-1) was not detected by co-mitogenic thymocyte assays of crude supernates. Isoelectrofocusing of concentrated medium showed tha...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90266-4
更新日期：1989-09-01 00:00:00
abstract：:Highly sensitive and easy detection method for Alzheimer's disease (AD) with a suitable biomarker is mandatory for preventing the factors resulting from AD. This research reports a modified ELISA with graphene for the detection of AD biomarker amyloid beta (Aβ) oligomer. Gold nanoparticle (AuNP) conjugated aptamer was...
journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.11.008
更新日期：2004-02-15 00:00:00
abstract：:There has been recent interest in the depletion of regulatory T cells (Tregs) as part of a multi-faceted approach to the immunotherapy of melanoma patients. This is in part due recent findings that convincingly show that Tregs are an integral part of regulating and even suppressing an immune response to growing tumor ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.01.012
更新日期：2008-04-20 00:00:00
abstract：:An indirect rosette assay which utilizes erythrocytes coupled with purified antiimmunoglobulin (Ig) antibodies was modified into a microassay for detecting HLA allo-and xenoantibodies. The test, which is performed in microtiter plates, is specific, reproducible and can handle large numbers of samples. As an inhibition...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90059-0
更新日期：1979-01-01 00:00:00
abstract：:Phagocytosis plays a pivotal and essential role in host immune defense, both as a focal constituent of the innate immune response and a bridging element linking innate and adaptive immunity. Phagocytosis has been demonstrated to be critical in development, tissue remodeling, wound healing and resolution of inflammatio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.027
更新日期：2013-04-30 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90353-8
更新日期：1983-05-27 00:00:00
abstract：:Mouse models of allergic asthma are increasingly used to study the immunopathology of this complex disorder. The degree and type of airway inflammation is often studied by determination of differential cell counts on cytospins of bronchoalveolar lavage fluid (BALF) cells stained with May-Grünwald Giemsa, in which the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.03.004
更新日期：2004-05-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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abstract：:Drugs that target tumor necrosis factor-alpha (TNF) are particularly important in the treatment of severe inflammatory progression in rheumatoid arthritis, Crohn's disease and psoriasis. Despite the central role of the TNF/TNF receptor (TNFR) in various disease states, there is a paucity of information concerning TNFR...
journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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doi：10.1016/j.jim.2018.04.007
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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abstract：:A relatively simple, specific and sensitive radioimmunoassay system has been developed for the detection of heterophile Hanganutziu-Deicher (H-D) antigen(s) and antibodies. The 125I-labeled H-D antigen-active molecule used for the assay is a bovine erythrocyte major glycoprotein previously found to have a strong H-D a...
journal_title：Journal of immunological methods
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abstract：:Immunoabsorbents for the removal of antibodies against cell membrane antigens have been prepared by a variety of techniques in common use. Cells were surface-labelled by lactoperoxidase-catalysed radioiodination, and the different immunoabsorbent preparations were compared in terms of efficiency of incorporation and s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90222-3
更新日期：1978-01-01 00:00:00
abstract：:A quantitative immunocytochemical method for the measurement of islet cell cytoplasmic antibodies has been developed. The method employs human or rat pancreas, a protein A-peroxidase/diaminobenzidine secondary antibody system and an independent measurement of islet total and exocrine mean integrated absorbance by scan...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(12)80049-4
更新日期：1992-04-27 00:00:00
abstract：:We describe here a lateral flow-based assay (LFA) for the detection of antibodies against immunodominant antigen Pgp3 from Chlamydia trachomatis, the causative agent of urogenital chlamydia infection and ocular trachoma. Optimal signal detection was achieved when the gold-conjugate and test line contained Pgp3, creati...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.05.008
更新日期：2016-08-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：2010-04-15 00:00:00
abstract：: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 effic...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.05.007
更新日期：2018-10-01 00:00:00
abstract：:We examined two ELISA methods for measuring antibodies to Streptococcus pneumoniae using tyraminated S. pneumoniae polysaccharide types 3, 7N, 9F and 14 as antigens. The ELISA has the usual format with a relatively long incubation time whereas the FAST-ELISA has a short incubation time and employs a different solid-ph...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90238-x
更新日期：1989-06-21 00:00:00
