Abstract：
:A flow cytometry-based assay for analyzing cytotoxic T lymphocyte (CTL) activity is presented. This new approach is characterized by easy handling, the generation of highly reproducible data sets and is not dependent on the use of radioactivity. Before exposure to primed CTL effector cells the target cells were labeled with the green fluorescent dye DiO18(3) which is incorporated stably into the cell membrane. After a 4-h incubation period, samples were counterstained with the red fluorescent nuclear dye propidium iodide in order to permit discrimination between live and dead cells within both cell populations. The assay has been used to quantitate CTL effector activity against allogeneic lymphoblasts. Results derived from this novel flow cytometry assay show an excellent correlation (r = 0.988) with data obtained using the standard 51chromium release assay. An additional advantage of the assay is that freshly prepared splenocytes may be used as target cells because culturing and activation of target cells is no longer required. The results demonstrate that the fluorescent dyes DiO18(3) and propidium iodide in combination with flow cytometry permit accurate analysis of cytotoxic T cell activity.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Mattis AE,Bernhardt G,Lipp M,Förster Rdoi
10.1016/s0022-1759(97)00047-1subject
Has Abstractpub_date
1997-05-26 00:00:00pages
135-42issue
2eissn
0022-1759issn
1872-7905pii
S0022-1759(97)00047-1journal_volume
204pub_type
杂志文章abstract：:A microELISA for the estimation of human serum thymosin alpha 1 is described. In this assay, antibody to thymosin alpha 1 is pre-incubated with the standard or serum at 4 degrees C. Unbound antibody in the liquid-phase then binds with solid-phase thymosin alpha 1. The method is sufficiently sensitive for measuring ser...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90163-2
更新日期：1985-06-12 00:00:00
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 ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.112642
更新日期：2019-11-01 00:00:00
abstract：:The in vitro migration inhibition responses of peripheral blood leukocytes from tuberculin skin test positive and negative donors were tested to validate and determine optimal conditions for the agarose droplet method in the human. In vitro migration inhibition was observed in skin test positive donors in the presence...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90063-7
更新日期：1978-01-01 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：:Cell hybridization techniques have already been utilized, but the outcome is usually not very stable. In order to achieve high stability we tried to achieve closer intercellular contact. Neuraminidase treatment was used to remove sialic acid from the cell membrane, since sialic acid usually prevents close contact betw...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90260-3
更新日期：1990-12-31 00:00:00
abstract：:A polyethylene glycol (PEG) radioimmunoprecipitation assay for human IgG is described that is sufficiently sensitive to detect 0.5 ng of IgG. This model antibody-antigen system was also used to study the stoichiometries of PEG-precipitated complexes. Our results suggest that the presence of PEG may affect the stoichio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90091-7
更新日期：1979-01-01 00:00:00
abstract：:We have developed a large scale (10 g/batch) non-chromatographic method for the collection of IgM directly from plasma obtained from therapeutic plasmapheresis of patients. The technique uses sequential precipitations from ammonium sulfate and polyethylene glycol, and gives high yields of undenatured IgM antibodies. T...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90252-3
更新日期：1993-01-14 00:00:00
abstract：:Immune complexes have been implicated in the pathogenesis of a number of infectious diseases. The predominant immunoglobulin class associated with circulating immune complexes is IgG, although immune complexes containing IgM have been described. The role of IgM immune complexes in disease pathogenesis has been difficu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(87)80005-4
更新日期：1987-08-24 00:00:00
abstract：:Preclinical models of lymph node (LN) metastasis are fundamental to the study and design of new techniques for the diagnosis and treatment of LN metastasis. However, the identification of LNs and lymphatic vessels (LVs) in mice is challenging with conventional imaging modalities, since the LN diameter in normal mice i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.01.004
更新日期：2013-03-29 00:00:00
abstract：:Frozen stored human mononuclear cells were compared with non-frozen cells as responder cells in the monocyte chemotaxis assay. Frozen stored cells were found to inhibit lower background migration, resulting in greater experimental-to-control ratios. The use of frozen cells also greatly improved the speed, standardizat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)90023-5
更新日期：1977-01-01 00:00:00
abstract：:Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to evaluate bacterial ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.12.004
更新日期：2019-02-01 00:00:00
abstract：:A simple apparatus for measuring chemotaxis is described which contains thirty chambers per unit. Only very few cells and small volumes of chemotactic factor solution are required per chamber. For satisfactory results the apparatus must be centrifuged briefly prior to incubation. Under these conditions results can be ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)90008-9
更新日期：1977-01-01 00:00:00
abstract：:A novel enzyme-linked bio-immunoassay (bio-ELISA) has been developed to detect interferon-gamma (IFN-gamma) induced HLA-DR antigen on the surface of human tumor cells. Cells are cultured at 37 degrees C in 96-well microtiter plates in the presence of IFN-gamma for 2 days. After fixation with reagent alcohol the HLA-DR...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90083-5
更新日期：1989-12-20 00:00:00
abstract：:ADP-ribosylation factors (Arf) are small GTP-binding proteins involved in vesicular transport and the activation of phospholipase D (PLD). The conversion of Arf-GDP to Arf-GTP is promoted in vivo by guanine nucleotide exchange factors such as ARNO or cytohesin-1. In order to examine the expression of ARNO and cytohesi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00336-7
更新日期：2001-03-01 00:00:00
abstract：:The T cell receptor (TCR) determines the cellular response to antigens, which are presented on the surface of target cells in the form of a peptide bound to a product of the major histocompatibility complex (pepMHC). The response of the T cell depends on the affinity of the TCR for the pepMHC, yet many TCRs have been ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.09.016
更新日期：2008-12-31 00:00:00
abstract：:An immunoaffinity method for the purification of hepatitis B surface antigen (HBsAg) using monoclonal anti-idiotype (anti-Id) as a specific eluent is described. Ammonium sulfate fractions of HBsAg-containing plasma were absorbed on monoclonal idiotype (Id) immunoadsorbents. The bound material was eluted from the colum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90019-r
更新日期：1990-01-24 00:00:00
abstract：:This report describes the use of biotin-labeled antibodies in sensitive enzyme immunoassay systems for the measurement of bacterial antigens. Biotinylated immunoglobulins could be reproducibly formulated using N-hydroxysuccinimide biotin ester to link biotin to the immunoglobulin. Binding of the biotinylated antibody ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(83)80021-0
更新日期：1983-02-11 00:00:00
abstract：:A solid-phase immunoradiometric assay is described for the detection of mouse immunoglobulin classes and subclasses in unpurified and unconcentrated supernatants of hybridomas. IgG fractions from rabbit antisera specific for mouse immunoglobulin classes and subclasses are used for coating the wells of flexible microti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90161-3
更新日期：1984-03-30 00:00:00
abstract：:Relatively purified populations of helper T cells free of active PFC precursors can be prepared from primed splenocytes by irradiation, hypotonic shock, and filtration over glass wool. These purified populations can be stored and helper activity recovered undiminished after up to 6 months of cryopreservation. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90082-0
更新日期：1978-01-01 00:00:00
abstract：:The sensitivity of any immunoassay is a complex function of the underlying physico-chemical basis of the technique and the size and source of 'experimental' errors. Analysis of this relationship emphasizes the distinction between competitive labelled analyte methods, e.g., RIA, and the non-competitive labelled antibod...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90338-8
更新日期：1986-02-27 00:00:00
abstract：:Interleukin 2 (IL-2) is a class of T cell growth factors produced by T cells of a number of species. The unique growth-promoting properties of these molecules allow the development of antigen-specific effector T cell lines which can be used to analyze the molecular basis of lymphocyte interactions. A murine T cell tum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90055-1
更新日期：1980-01-01 00:00:00
abstract：:A simple method to study 19S anti-SRBC antibody production is described. This technique, performed by incubation of immune spleen cells, SRBC and C in the fluid phase, utilises photomeric estimation of antibody mediated haemoglobin release. The test is quicker and more accurate than the two-step haemolytic plaque assa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90143-0
更新日期：1977-01-01 00:00:00
abstract：:Testing of diphtheria vaccines for routine lot release relies heavily on the use of in vivo potency assays. However, consistency of production is also recognised as an important feature of vaccine manufacture, and in vitro assays are superior to in vivo assays for providing this information. In adsorbed vaccines, anti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.09.002
更新日期：2009-10-31 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
更新日期：2016-04-01 00:00:00
abstract：:Since the number of polymorphonuclear leukocytes (PMN) responding to a chemotactic stimulus in the Boyden chamber is influenced by the cell density, we studied whether this variable was important in determining chemotaxis under agarose. The chemotactic index was determined by summing the product of each cell that had ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90292-1
更新日期：1981-01-01 00:00:00
abstract：:An enzyme-linked immunosorbent assay was developed for quantitation of complement C3b receptors (C3bR) on human erythrocytes fixed in monolayer to microtiter plates. The disadvantages of macro test tube systems (large consumption of sample material and reagents, tedious washing procedures, cell loss and hemolysis) wer...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90211-x
更新日期：1985-08-02 00:00:00
abstract：:The multiple antigen peptide (MAP) system has been proposed as a novel and valuable approach for eliciting antibodies to peptides and developing synthetic vaccines. The MAP system consists of a small immunogenically inert core matrix of lysine residues with alpha- and epsilon-amino groups for anchoring multiple copies...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90033-p
更新日期：1992-12-08 00:00:00
abstract：:Bromodeoxyuridine (BrdU) is used to label synthesizing DNA and to chase label-retaining cell (LRC). As stem cells divide slowly in adult tissues, they can be visualized as LRCs. In order to identify LRCs in hard tissue, we examined optimal conditions of fixation, demineralization, and DNA denaturation/antigen retrieva...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.07.013
更新日期：2008-11-30 00:00:00
abstract：:We have developed and validated an inexpensive and equivalent method for measuring eosinophil adhesion by beta(2)-integrin to endothelial ICAM-1 using bovine serum albumin (BSA) as a surrogate for the immunoglobulin supergene. The number of adherent eosinophils on BSA or ICAM-1 coated microplates was quantified by res...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00192-7
更新日期：2000-06-23 00:00:00
abstract：BACKGROUND:The serological screening for celiac disease (CD) is currently based on the detection of anti-transglutaminase (tTG) IgA antibodies, subsequently confirmed by positive endomysial antibodies (EMA). When an anti-tTG IgA positive/EMA IgA negative result occurs, it can be due either to the lower sensitivity of t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112910
更新日期：2021-02-01 00:00:00
