Multiparametric flow cytometric analysis of biochemical and functional events associated with apoptosis and oncosis using the 7-aminoactinomycin D assay.

Abstract:

:Apoptosis and primary necrosis are the two modes of cell death induced by a lethal injury. The majority of structural and biochemical events occurring during cell death can be analysed by flow cytometry. The 7-aminoactinomycin D (7-AAD) assay can be used to detect the loss of membrane integrity during apoptosis of murine thymocytes and human peripheral lymphocytes. We describe here new applications of the 7-AAD assay. It can be applied to a variety of cell lines of different origins, including adherent cell lines, and it allows the co-detection of lipidic antigens such as phosphatidylserine (PS) residues, and biochemical processes linked to apoptosis, such as the loss of mitochondrial transmembrane potential, cardiolipin peroxidation, the expression of the 7A6 mitochondrial antigen and DNA fragmentation. Thus, this assay is a noninvasive method particularly adapted to the analysis of biochemical events associated with cell death. Finally, we show that this assay is not specific for apoptosis since it detects oncosis, the early stage of primary necrosis.

journal_name

J Immunol Methods

authors

Lecoeur H,de Oliveira-Pinto LM,Gougeon ML

doi

10.1016/s0022-1759(02)00072-8

keywords:

subject

Has Abstract

pub_date

2002-07-01 00:00:00

pages

81-96

issue

1-2

eissn

0022-1759

issn

1872-7905

pii

S0022175902000728

journal_volume

265

pub_type

杂志文章,评审
  • Indirect 125I-labeled protein A assay for monoclonal antibodies to cell surface antigens.

    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

    authors: Brown JP,Tamerius JD,Hellström I

    更新日期:1979-01-01 00:00:00

  • A technique for determining E rosette levels by cytofluorographic analysis.

    abstract::A method is described for determining the percentage of rosettes formed by sheep erythrocytes and human peripheral lymphocytes (ER) using cytofluorographic analysis (CFGA). The technique utilizes acridine orange dye for differentially staining nucleus and cytoplasm of the lymphocytes to distinguish them from erythrocy...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(76)90091-0

    authors: Goldberg NH,Kenady DE,Super BS,Chretien PB

    更新日期:1976-01-01 00:00:00

  • Rapid screening of monoclonal antibodies: new 'microstick' radioimmunoassay.

    abstract::A new system for assaying monoclonal antibodies consisting of an 8 x 12 array of sticks which fits into a 96-well microtiter plate is described. Tests using virus specific monoclonal antibodies and virus proteins demonstrated sensitivity equivalent to the conventional microtiter plate assay. Antibody production, antig...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90356-3

    authors: Scheinberg DA,Pan XQ,Wilsnack R,Strand M

    更新日期:1983-03-25 00:00:00

  • Production of a monoclonal antibody reactive with poly(A) and its application in the isolation of poly(A)+ RNA.

    abstract::Hybridoma cells producing antibody reactive with poly(A) were isolated following fusion of spleen cells from a BALB/c mouse immunized with poly(A).poly(dT) and myeloma cells. The antibody was purified from ascitic fluid by the formation of immune complexes with poly(A). The antibody reacted with poly(A) and some homop...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90269-d

    authors: Kitagawa Y,Ishibashi K

    更新日期:1993-08-26 00:00:00

  • Efficient display of scFv antibodies on bacteriophage lambda.

    abstract::In the present work we demonstrate the efficient display of functional scFv antibodies on the bacteriophage lambda capsid. A single-chain (scFv) anti-CEA antibody gene was cloned in two different vectors to obtain fusion of the scFv antibody to the N- or C-terminus of the bacteriophage lambda capsid protein D (gpD). L...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2006.01.003

    authors: Vaccaro P,Pavoni E,Monteriù G,Andrea P,Felici F,Minenkova O

    更新日期:2006-03-20 00:00:00

  • A rapid FPLC method for purification of the third component of human and guinea pig complement.

    abstract::A method is described for the purification of human and guinea pig C3 from small amounts of serum. This procedure requires only two steps--polyethylene glycol (PEG) precipitation and fast protein liquid chromatography (FPLC) Mono Q HR 10/10 ion exchange chromatography. The protocol takes less than two hours to complet...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90290-v

    authors: Basta M,Hammer CH

    更新日期:1991-08-28 00:00:00

  • ELISA methods for the analysis of antibody responses induced in multiple sclerosis patients treated with recombinant interferon-beta.

    abstract::Upon treatment with protein therapeutics, a subset of patients will typically develop antibodies against the drug. These anti-drug antibodies can be of concern because they have the potential to alter the drug's therapeutic activity. In the case of relapsing-remitting multiple sclerosis (RRMS) patients receiving recom...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(99)00073-3

    authors: Brickelmaier M,Hochman PS,Baciu R,Chao B,Cuervo JH,Whitty A

    更新日期:1999-07-30 00:00:00

  • Detection of circulating immune complexes with a Raji cell enzyme immunoassay.

    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

    authors: Rote NS,Caudle MR

    更新日期:1983-01-01 00:00:00

  • Production and characterisation of monoclonal antibodies to parathyroid hormone-related protein.

    abstract::The production and characterisation of 17 monoclonal antibodies to human parathyroid hormone-related protein (PTH-rP) 1-34 is described. Five of the antibodies were shown to be of high avidity (Ka 4 X 10(10)-1.9 X 10(11) L/M) and able to detect 15-100 pg PTH-rP 1-34 per tube by RIA. None cross-reacted with PTH 1-34, a...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(90)90346-w

    authors: Ratcliffe WA,Hughes S,Gilligan MG,Heath DA,Ratcliffe JG

    更新日期:1990-02-20 00:00:00

  • Hepatocyte growth factor reverses the TGF-beta-induced growth inhibition of CCL-64 cells. A novel bioassay for HGF and implications for the TGF-beta bioassay.

    abstract::The influence of human hepatocyte growth factor (HGF) on the transforming growth factor beta (TGF-beta) bioassay CCL-64 was examined. HGF induced proliferation of the CCL-64 cells and potently counteracted TGF-beta-induced growth inhibition. HGF was not inactivated by transient acidification to pH 2, a commonly used p...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(95)00228-6

    authors: Börset M,Waage A,Sundan A

    更新日期:1996-01-16 00:00:00

  • A modified screening method for pcDNA-1 expression libraries which is applicable to both surface and intracellular antigens. Cloning of a colon carcinoma antigen.

    abstract::A modification of the COS cell transient expression system for isolating cDNAs encoding antibody defined molecules was used to clone the carcinoma antigen defined by monoclonal antibody CORC-6. Bacteria were transformed with cDNA prepared from colon carcinoma cells and immediately divided into aliquots containing appr...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(95)00155-4

    authors: Grimm T,Johnson JP

    更新日期:1995-10-26 00:00:00

  • Estimation of low frequency antigen-presenting cells with a novel RELISPOT assay.

    abstract::Adequate presentation of self and foreign antigens is a key factor for efficient T-cell immunosurveillance against pathogens and tumors. Cells presenting foreign antigens usually comprise a rare population and are difficult to detect even at the peak of infection. Here we demonstrate a CD8(+) T-cell-based approach tha...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2008.01.008

    authors: Dzutsev AK,Belyakov IM,Isakov DV,Gagnon SJ,Margulies DH,Berzofsky JA

    更新日期:2008-04-20 00:00:00

  • Scanning tunnelling microscopy and dynamic contact angle studies of the effects of partial denaturation on immunoassay solid phase antibody.

    abstract::A range of partial denaturation antibody pre-treatments that affect immunoassay performance have been evaluated. Monoclonal anti-ferritin antibody was either partially denatured by heat, urea or pH pre-treatment or left untreated and then passively adsorbed to polystyrene microtiter wells. The adsorption characteristi...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(95)00141-v

    authors: Davies J,Dawkes AC,Haymes AG,Sunderland RF,Edwards JC

    更新日期:1995-10-12 00:00:00

  • In vitro assessment of delayed hypersensitivity in the human. Inhibition of cell migration from agarose microdroplets.

    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

    authors: Lavergne JA,Harrington JT

    更新日期:1978-01-01 00:00:00

  • The in vitro production of chimeric murine thymus from nonlymphoid embryonic precursors.

    abstract::A simplified technique for the production of chimeric thymus completely in vitro from the third pharyngeal pouch (TPP) of 10-day mouse embryos has been examined. The results are similar to those obtained by a more cumbersome technique published by others. This modified technique permits the production of large numbers...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90279-x

    authors: Pyke KW,Bartlett PF,Mandel TE

    更新日期:1983-03-11 00:00:00

  • Detection of human in vitro hapten specific antibody responses.

    abstract::A microculture and radioimmunoassay is described which is capable of generating and detecting very small levels of human anti-DNP antibody. The antibody produced is hapten specific and the response is T-cell dependent. The microculture system described avoids the need for high serum supplements, mitogens, in vivo immu...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90322-8

    authors: Macdonald PR,Simpson EJ,Crosier PS

    更新日期:1983-12-16 00:00:00

  • Immunoabsorbents prepared from cell membrane antigens: efficiency of incorporation and shedding of protein during use.

    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

    authors: Zola H

    更新日期:1978-01-01 00:00:00

  • Application of a fluorometric assay to detect caspase activity in thymus tissue undergoing apoptosis in vivo.

    abstract::To date, in vivo apoptosis within the thymus has been assessed using morphological criteria and/or detection of a DNA ladder indicative of oligonucleosomal fragmentation of the DNA. Here, we have used a fluorometric method to investigate activation of the caspase protease family in the thymus following in vivo inducti...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(99)00054-x

    authors: Gorman AM,Hirt UA,Zhivotovsky B,Orrenius S,Ceccatelli S

    更新日期:1999-06-24 00:00:00

  • Methods for generation of monoclonal antibodies to the very small drug hapten, 5-benzimidazolecarboxylic acid.

    abstract::Drug-specific monoclonal antibodies (MAbs) were produced against the very small drug hapten (162.15 Da), 5-benzimidazolecarboxylic acid, an analogue of 2-(4-Thiazolyl)benzimidazole (TBZ) but lacking the thiol group. TBZ is widely used as a broad-spectrum anthelmintic in various animal species and humans and also as a ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(02)00340-x

    authors: Moran E,O'Keeffe M,O'Connor R,Larkin AM,Murphy P,Clynes M

    更新日期:2002-12-20 00:00:00

  • Comparison of highly NK active human lymphocyte subsets separated by various procedures involving E, EA rosetting, density gradients and adherence to immune complexes.

    abstract::Lymphocyte subsets from human blood obtained by different procedures were analyzed for cytotoxic potential and phenotypic characteristics. Nylon wool column passed lymphocytes were fractionated on the basis of: (1) E and Fc gamma receptor expression, (2) cell density and Fc gamma receptor expression, (3) Fc gamma rece...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90209-0

    authors: Masucci G,Masucci MG,Bejarano MT,Klein E

    更新日期:1983-09-30 00:00:00

  • Antigen leakage from immunosorbents. Implications for the detection of site-directed auto-anti-idiotypic antibodies.

    abstract::The detection of site-directed anti-idiotypic antibodies is usually based on their ability to inhibit the binding of antigen to idiotype, in either solid- or fluid-phase radioimmunoassays. Passage of serum over antigen-coupled immunosorbents for the purpose of removing the idiotypes from complexes with putative anti-i...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(87)90172-4

    authors: Hagen M,Strejan GH

    更新日期:1987-06-26 00:00:00

  • The use of supernatants from neuraminidase and galactose oxidase (NAGO)-treated lymphocytes as a source of T cell growth factor.

    abstract::Human peripheral blood or tonsil lymphocytes produce T cell growth factor (TCGF), when activated with neuraminidase (NA) and galactose oxidase (GO). Partial purification of NAGO-TCGF on Sepharose G-100 columns gave a TCGF-active fraction within the same molecular weight range as the conventional lectin-induced TCGF (a...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(83)80004-0

    authors: Berzins T,Wikén M,Hellström U,Perlmann P

    更新日期:1983-10-28 00:00:00

  • Leukopak PBMC sample processing for preparing quality control material to support proficiency testing programs.

    abstract::External proficiency testing programs designed to evaluate the performance of end-point laboratories involved in vaccine and therapeutic clinical trials form an important part of clinical trial quality assurance. Good clinical laboratory practice (GCLP) guidelines recommend both assay validation and proficiency testin...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2014.05.019

    authors: Garcia A,Keinonen S,Sanchez AM,Ferrari G,Denny TN,Moody MA

    更新日期:2014-07-01 00:00:00

  • Measurement of IL-21 in human serum and plasma using ultrasensitive MSD S-PLEX® and Quanterix SiMoA methodologies.

    abstract::IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commer...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2018.12.005

    authors: Poorbaugh J,Samanta T,Bright SW,Sissons SE,Chang CY,Oberoi P,MacDonald AJ,Martin AP,Cox KL,Benschop RJ

    更新日期:2019-03-01 00:00:00

  • A general and mild procedure for the purification of rabbit Fab' antibodies.

    abstract::A mild procedure for the purification of rabbit Fab' antibodies is described. A small column of normal goat IgG-Sepharose 4B (3 cm x 5mm) was saturated with F(ab')2 prepared from rabbit anti-goat IgG, and anti-goat IgG Fab was eluted from the column by splitting the disulfide bond in the hinge of the F(ab')2 molecule ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(80)90336-1

    authors: Ishikawa E,Yoshitake S

    更新日期:1980-01-01 00:00:00

  • Fluorescent target array T helper assay: a multiplex flow cytometry assay to measure antigen-specific CD4+ T cell-mediated B cell help in vivo.

    abstract::CD4(+) T cells play a central role in regulating the immune response. Their effector function is commonly assessed by their capacity to secrete cytokines detected by ELISPOT and intracellular cytokine staining. However, one aspect of their effector function that is often overlooked is their ability to help activation ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2012.10.013

    authors: Quah BJ,Wijesundara DK,Ranasinghe C,Parish CR

    更新日期:2013-01-31 00:00:00

  • Difficulties in obtaining monoclonal antibodies to subsets of human leukocytes, using neonatal tolerance induction in mice.

    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

    authors: Yamaguchi Y,O'Doherty U,Peng M,Steinman RM

    更新日期:1995-04-12 00:00:00

  • An enzyme-linked immunosorbent assay (ELISA) for the determination of diphtheria toxin antibodies.

    abstract::An enzyme-linked immunosorbent assay (ELISA) for the quantitation of diphtheria toxin antibodies is described. When the ELISA technique was compared to a single radial immunodiffusion assay the results correlated well but the ELISA technique was ten thousand times more sensitive. It was also at least ten times as sens...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(77)90107-7

    authors: Svenson SB,Larsen K

    更新日期:1977-01-01 00:00:00

  • Discovery of biomarkers for systemic lupus erythematosus using a library of synthetic autoantigen surrogates.

    abstract::Antibodies to a wide range of self-antigens, including those directed against nucleic acids or nucleic acid-binding proteins are the essential biomarkers for diseases such as systemic lupus erythematosus (SLE). Highly complex libraries of nonamers consisting of N-substituted glycines (peptoids) were screened for compo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2013.11.004

    authors: Quan J,Lakhanpal A,Reddy MM,Zaman S,Li QZ,German DC,Olsen NJ,Kodadek T,Karp DR

    更新日期:2014-01-15 00:00:00

  • Optimization of the modification of carrier proteins with aminated haptens.

    abstract::In this report we show that succinic groups are far more reactive to amino compounds than the carboxylic groups derived from Asp and Glu on the protein when using coupling via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDCI) (even by an 8 fold factor). Accordingly, a new carrier-protein was designed where both na...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2005.10.003

    authors: Fuentes M,Palomo JM,Mateo C,Venteo A,Sanz A,Fernández-Lafuente R,Guisan JM

    更新日期:2005-12-20 00:00:00