Abstract：
:Goat anti-mouse antibodies conjugated with colloidal 40 nm gold particles (G40) were used as secondary layers to stain human T lymphocytes in an attempt to extend the polarized light epi-illumination microscopic technique to flow cytometric multiparameter analysis. G40-labelled T cells were further stained with phycoerythrin (PE, red)- and fluorescein (FITC, green)-conjugated antibodies with specificity for the same cells. The cell samples were then analysed on a standard flow cytometer equipped with one laser operating at 488 nm. G40-labelled cells were detected in the side scatter (90 degrees) channel and fluorescent cells in the red and green fluorescence channels, respectively. Single labelling of the same T cell subsets with either G40-, FITC- or PE-conjugated antibodies yielded similar results, and cell mixture experiments did not show interference between gold and fluorescence labels. Triple staining experiments with three differently conjugated antibodies showed that subpopulations of cells were labelled in an independent manner with one, two or three antibodies in proportions expected from fluorescence controls. It was, therefore, possible to detect subset, sub-subset and activation markers on individual T cells. Our experiments show that immunogold staining of cell populations can be detected in routine one-laser flow cytometry. Further, the gold label can be combined to give two-color staining with ordinary red and green fluorochrome-conjugated antibodies thereby permitting rapid and precise triple antibody staining of individual cells. This methodology provides a powerful tool for the analysis of cellular antigenic diversity among, e.g., immunocompetent cells.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Festin R,Björklund B,Tötterman THdoi
10.1016/0022-1759(87)90211-0subject
Has Abstractpub_date
1987-07-16 00:00:00pages
23-8issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(87)90211-0journal_volume
101pub_type
杂志文章abstract：:The short-term kinetics of T cell-mediated cytotoxicity was investigated using a cloning inhibition assay. Murine cytotoxic thymus-derived lymphocytes (CTL) generated in vitro in mixed leukocyte cultures (MLC), were incubated for various periods of time at 37 degrees C with allogeneic mastocytoma target cells. The mix...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90201-0
更新日期：1977-01-01 00:00:00
abstract：:A new, relatively simple, spectrophotometric technique has been developed which is useful for accurately determining the extent of chromophore labeling of proteins. Often the absorbance spectra and extinction coefficients of dye/protein conjugates are strongly affected by changes in the chromophore microenvironment th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90051-g
更新日期：1991-10-25 00:00:00
abstract：:A sensitive assay, using [14C]lecithin as a substrate, has been developed for the measurement of phospholipase activity in rat peritoneal polymorphonuclear leukocytes. Cell extracts were found to contain a phospholipase D activity and indirect evidence suggested that eosinophils are responsible for the cleavage of lec...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90212-4
更新日期：1980-01-01 00:00:00
abstract：:Toll-like receptor (TLR) agonists are currently being examined as adjuvants for vaccines, with several lead candidates now in licensed products or in late-stage clinical development. Guinea pigs are widely used for preclinical testing of drugs and vaccines; however, evaluation of TLR agonists in this model is hindered...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.07.021
更新日期：2011-10-28 00:00:00
abstract：:A lectin isolated from the tropical jackfruit, jacalin, previously reported to precipitate human immunoglobulin A (IgA), and conjugated to agarose was used to separate the two subclasses of IgA from secretions. Jacalin-agarose binds specifically to the D-galactose moiety of IgA1 but not to IgA2 which has a different c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90037-8
更新日期：1987-05-04 00:00:00
abstract：:Simplified isolation procedures are described for the parenchymal cell of the liver and the major non-parenchymal cell, the Kupffer cell. Hepatocytes are obtained in a purity of approximately 100%; a yield 10 X 10(6) cells/g liver tissue and the viability is greater than 85%. The recovery of Kupffer cells is 82%, viab...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90262-x
更新日期：1979-01-01 00:00:00
abstract：OBJECTIVE:The objective of this study was the identification of the stain HIF-alpha using the Image Cytometry, and to help to count the positive cells (with HIF-alpha) and the negative cells (without HIF-alpha) from the same sample. METHOD:17 images of renal tissues from male rats of Winstar lineage; overall, there we...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.02.005
更新日期：2017-05-01 00:00:00
abstract：:An assay for detection of monoclonal hybridoma antibodies against cell surface antigens is described. Samples of spent medium from the hybridoma cultures are incubated in microtest wells with cells, either as adherent monolayers or in suspension. Antibodies bound to surface antigens are detected by successive incubati...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90132-7
更新日期：1979-01-01 00:00:00
abstract：:Dendritic Cell (DC)-based vaccination approaches in man require a reproducible DC generation method that can be performed in conformity with GMP (Good Manufacturing Practice) guidelines and that circumvents the need for multiple blood drawings to generate DC. To this end we modified our previously described method to ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(98)00208-7
更新日期：1999-02-01 00:00:00
abstract：:Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-in...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00201-5
更新日期：2000-08-28 00:00:00
abstract：:AMPKalpha is a subunit of AMP-activated protein kinase (AMPK), a heterotrimeric enzyme that works as a fuel sensor activated in response to the depletion of cellular ATP. AMPKalpha is considered as a master switch in regulating glucose and lipid metabolism. Determining its presence in patient sera may help in diagnosi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.10.001
更新日期：2009-12-31 00:00:00
abstract：:Two methods for estimating IgA in body fluids of rats were devised and tested for their accuracy and reliability. Specifically purified monomeric and polymeric IgA was prepared so that results obtained could be properly assessed. The two systems tried were prepared so that results obtained could be properly assessed. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90293-x
更新日期：1979-01-01 00:00:00
abstract：:The reactivity of monoclonal antibodies (Mabs) raised against a cyclic peptide representing a chimeric V3 loop of HIV-1 gp120 with different peptide analogues was studied with a biosensor system (BIAcore) and by ELISA. In both assays, the Mabs cross-reacted extensively with the V3 regions of different HIV-1 strains an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90316-6
更新日期：1994-12-02 00:00:00
abstract：:The Raji cell assay for circulating immune complexes (CIC) is frequently the method of choice when the detection of large, complement fixing complexes is desired. We have developed an enzyme-linked immunosorbent assay modification (Raji-ELISA) of the Raji cell technique which is easy to perform, uses commercially avai...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90046-7
更新日期：1983-01-01 00:00:00
abstract：:A cell ELISA for human leukocytes has been established and standardized. The assay was based on the use of air-dried and methanol-fixed cells which were attached to microplate wells. Cellular antigens were measured through the subsequent binding of monoclonal antibody (MoAb), biotinylated sheep Ig against mouse IgG an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90219-x
更新日期：1986-11-20 00:00:00
abstract：:Cut point determination is an important aspect of immunogenicity assay development. The cut point can be influenced by a myriad of factors. Key among those is the analytical variability of the assay itself and biological variation due to test samples. Since a smaller cut point value may result in improved sensitivity,...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.10.001
更新日期：2018-12-01 00:00:00
abstract：:A new type of continuous, supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing specific fractionation and separation of human peripheral blood lymphocytes in a chromatographic format. The affinity adsorbent was used to design a novel cell separation strategy, which was based on the interact...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.09.017
更新日期：2003-12-01 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
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90049-5
更新日期：1989-03-10 00:00:00
abstract：:RIA and ELISA were compared for their ability to detect IgE in different rodent species. With a sheep anti-rat IgE antibody good correlation (P less than 0.001) between the 2 assay methods for IgE was found in rats. RIA failed to detect the IgE of Mastomys natalensis while ELISA proved to be a suitable test. However, ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90284-9
更新日期：1984-04-13 00:00:00
abstract：:Accurately predicting peptides binding to major histocompatibility complex (MHC) I molecules is of great importance to immunologists for elucidating the underlying mechanism of immune recognition and facilitating the design of peptide-based vaccine. Various computational methods have been developed for MHC I-peptide b...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.09.007
更新日期：2011-11-30 00:00:00
abstract：:A method for the production of human gamma interferon (HuIFN-gamma), which is both simple and efficient, has been developed. The 2 co-inducers, A-23187 and mezerein, stimulate human peripheral blood lymphocytes to produce high levels of HuIFN-gamma in both stationary cultures and spinner cultures. The compound 2,2-dim...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90428-3
更新日期：1983-10-14 00:00:00
abstract：:A panel of 18 monoclonal antibodies was raised to the human calcitonin gene related peptide (CGRP). Of these mabs, seven were specific for alpha CGRP and five for beta CGRP, while the remainder reacted with both alpha and beta CGRP. Nine different epitopes on CGRP were defined with these mabs. In addition, the mabs we...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90115-c
更新日期：1990-11-06 00:00:00
abstract：:The neutrophilic granulocyte is the most numerous leukocyte in peripheral blood. The development from a multipotent progenitor cell to a mature neutrophil takes place in the bone marrow over a period of 10-14 days. In order to understand the cellular mechanisms behind this process, it is necessary to investigate cells...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(99)00176-3
更新日期：1999-12-17 00:00:00
abstract：:To increase the selection efficiency and productivity of stable clones expressing recombinant antibody-IL-2 fusion protein, a dicistronic expression vector containing the anti-erbB2 scFv-Fc-IL-2 fusion gene followed by a weakened neo gene, was constructed to allow for the concurrent translation of the recombinant prot...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.08.013
更新日期：2004-12-01 00:00:00
abstract：:Interactions between mouse and rat monoclonal antibodies (MAbs) in EIA have been detected using panels of mouse and rat MAbs of various isotypes. Mouse and rat antibodies of the IgG2a subclass were found to bind to each other most strongly. Immobilised IgG1 antibodies of the two species showed much less interaction. R...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90300-k
更新日期：1990-06-12 00:00:00
abstract：:Antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action implicated in the clinical efficacy of several therapeutic antibodies. In vitro ADCC assays employing effector cells capable of inducing lysis of target cells bound by antibodies are routinely performed to support the research and...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2014.03.021
更新日期：2014-05-01 00:00:00
abstract：:The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. W...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.06.011
更新日期：2015-10-01 00:00:00
abstract：:Anti-sperm cell-mediated immunity (CMI) is considered as a crucial facet of infertility in patients of both sexes. A precise and objective method is designed, based on a one-step agarose leukocyte migration inhibition factor (LMIF) assay. The migration areas are evaluated by a computer-assisted image analysis system. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90186-w
更新日期：1992-10-02 00:00:00
abstract：:Natural CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) control the activation of the immune system and therefore have become a major area of research in immunology. The generation of monoclonal antibodies against human Tregs offers the possibility to discover novel Treg-specific or Treg-associated surface markers an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.01.002
更新日期：2010-02-28 00:00:00
abstract：:A microscale ELISA system, designated as the 'C1q-micro ELISA,' has been developed for the measurement of circulating immune complexes. The sensitivity of the method ranged between 0.1 and 3 microgram equivalents of aggregated human gamma-globulins. The procedure requires small quantities of serum and reagents, and it...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90308-8
更新日期：1982-01-01 00:00:00
