Abstract:
:The clinical value of assessing immunoglobulin (Ig)G and IgA subclasses in addition to the isotypes of soluble Igs in serum has been well established. >20years ago, the International Union of Immunological Societies and the World Health Organization performed collaborative studies in order to validate antibody (Ab) clones for the detection of IgG and IgA subclasses for a broad range of laboratory assays, except for flow cytometry. Here we analyzed the performance of commercially available Ab clones to detect IgG and IgA subclasses in memory B-cells and plasma cells (PCs) by flow cytometry. In a first step, 28 Ab clones were evaluated in peripheral blood from healthy donors. Only 17/28 clones showed reactivity against IgG and IgA subclasses expressed on the B-cell and PC surface membrane, including Ab clones for IgG1 (SAG1, HP6188, HP6001 and HP6186), IgG2 (SAG2, HP6014 and HP6002), IgG3 (SAG3, HP6095 and HP6050), IgG4 (SAG4), IgA1 (SAA1, H69-11.4 and B3506B4) and IgA2 (SAA2, 2E2, and A9604D2). In a second step, for each Ig subclass a single clone was selected according to its specificity and fluorescence intensity (resolution power), for further more detailed validation (SAG1, SAG2, SAG3, SAG4, SAA1 and SAA2). This validation process was carried out in 4 different laboratories by testing the selected Ab clones in human peripheral blood, bone marrow and tonsil samples, using different staining protocols (e.g. surface membrane and/or cytoplasmic staining). All selected Ab clones displayed strong positivity, high specificity and optimal resolution between negative and positive cells. Alternative Ab clones were also validated. Thus, our results show the feasibility of using the validated Ig subclass Ab clones in combination with other B cell-associated markers for detailed dissection of the memory B-cell and PC compartments that express distinct Ig subclasses in different human tissues.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Blanco E,Perez-Andres M,Sanoja-Flores L,Wentink M,Pelak O,Martín-Ayuso M,Grigore G,Torres-Canizales J,López-Granados E,Kalina T,van der Burg M,Arriba-Méndez S,Santa Cruz S,Puig N,van Dongen JJM,Orfao Adoi
10.1016/j.jim.2017.09.008subject
Has Abstractpub_date
2019-12-01 00:00:00pages
112372eissn
0022-1759issn
1872-7905pii
S0022-1759(17)30079-0journal_volume
475pub_type
杂志文章abstract::Dot-immunobinding assay for anti-calmodulin (CaM), and immunoblot assay for CaM have become feasible by proper choice of a fixative, i.e., formaldehyde vapor. The protocol for these simple and convenient immunological procedures is presented. Dot-immunobinding assay could be accomplished in 3 h, and immunoblot assay i...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90089-6
更新日期:1989-12-20 00:00:00
abstract:BACKGROUND:In light of the HIV pandemic, significant strides have been made in improving treatment options for patients. Technologies to monitor the progress of a patient on such treatment have therefore also been scaled up. Immune activation as measured by CD38 mean fluorescence intensity (MFI) on CD8 T cells has been...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.02.013
更新日期:2012-04-30 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::Antibodies directed against advanced glycosylation end products (AGEs) formed during a Maillard reaction have been generated and characterized. Since protein-bound AGEs recognized by the antibodies were labile to acid hydrolysis, the antibodies were further characterized by using the AGE-alpha-acetyl-L-lysine methyl e...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90133-z
更新日期:1991-06-24 00:00:00
abstract::Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90368-h
更新日期:1993-12-03 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::DNA microarray technology has developed rapidly in recent years and has become an essential tool, providing novel approaches to biomedical research. In this paper, we describe a self-designed ImmunoChip cDNA array for immunological research. With a comprehensive selection of genes of interest, we can focus on key sign...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2005.06.004
更新日期:2005-08-01 00:00:00
abstract::A simple two stage assay using latex particles as a reaction indicator has been developed for the detection of IgM antibodies to Trichinella spiralis. In the first stage, magnetic polystyrene beads (Dynabeads) coated with T. spiralis antigen were incubated for 30 min with the test serum. After washing, in the second s...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90149-x
更新日期:1989-02-24 00:00:00
abstract::We have developed a competitive enzyme immunoassay suitable for routine monitoring of intracellular levels of 5'-monophosphate-AZT (AZT-MP). This assay is performed in 96-well microtiter plates coated with anti-rabbit immunoglobulin antibodies and is based on the use of rabbit polyclonal antibodies raised against an A...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(98)00101-x
更新日期:1998-09-01 00:00:00
abstract::Two highly sensitive immunoassays for measuring soluble DNA are described. These methods employ a purified mouse monoclonal anti-DNA antibody in enzyme-linked immunoassays. One assay is an antigen capture method utilizing immobilized antibody. Bound DNA is subsequently detected with biotinylated monoclonal anti-DNA an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90005-0
更新日期:1986-04-17 00:00:00
abstract::A plasmid cassette vector was designed to generate transgenic mice expressing reporter cDNAs at high levels in antigen-presenting cells under the control of the murine invariant chain (Ii) promoter. Analysis of several transgenic mice harboring a chimeric Ii cDNA placed in this vector showed that it can drive expressi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00276-3
更新日期:2000-11-01 00:00:00
abstract::Real-time polymerase chain reaction (PCR) is now becoming an accepted tool for measuring gene expression at the transcriptional level. In this study, a direct comparison between real-time PCR, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunosorbent spot (ELISPOT) assay was performed. When interferon-...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(02)00029-7
更新日期:2002-05-01 00:00:00
abstract::This paper describes the application of experimental design techniques to optimize a sensitive ELISA for a hapten molecule with a calibration range of 0-1000 pg/ml. Ten factors that were expected to affect the assay performance were initially screened, followed by factorial experiments to delineate the effects of the ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00262-6
更新日期:1996-04-19 00:00:00
abstract::We describe a solid-phase fluoroimmunoassay, based on competition between europium-labeled 5 MeCyd (5-methylcytidine) and sample 5MedCyd (5-methyl-2'-deoxycytidine) for polyclonal anti-5MedCyd antibodies (rabbit). Europium labeling of antigen was performed using a novel polylysine-5MeCyd conjugate. Standard and sample...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90115-4
更新日期:1989-02-08 00:00:00
abstract::As a tetrameric major histocompatibility complex (MHC) class I-peptide complex (tetramer) is capable of detecting antigen-specific cytotoxic T lymphocyte (CTL) by flow cytometry, significant information about the generation of in vivo immunity can be obtained. It is, however, difficult to make a soluble wild type of M...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(02)00329-0
更新日期:2002-12-20 00:00:00
abstract::Four alleles are currently recognised at the HLA-DRB3 locus (DRB3*0101, DRB3*0201, DRB3*0202 and DRB3*0301). We studied whether 271 bp fragments of the polymorphic second exon, which were prepared using the polymerase chain reaction, could be typed using temperature gradient gel electrophoresis. Thermal stability curv...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)90063-9
更新日期:1994-02-10 00:00:00
abstract::Studies describing the induction of apoptosis for CD4 mAbs do not delineate between epitope-dependent and Fc-driven epitope cross-linking induced cell death. Keliximab and clenoliximab are two CD4 mAbs that differ only in their heavy chain isotypes, being an IgG1 and a modified IgG4, respectively. These antibodies sup...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00447-1
更新日期:2001-11-01 00:00:00
abstract::We wished to determine whether injection of mice with anti-isotype antibody would be a means to regulate in vivo isotype expression and to obtain hybridomas secreting monoclonal antibodies (mAbs) of desired antigen specificity and isotype. Treatment with a rat mAb (7D2) reactive with both the IgG2a and IgG2b isotypes ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90007-5
更新日期:1988-02-24 00:00:00
abstract::An indirect immunofluorescence technique was developed for the detection of antibodies to dsDNA and the simultaneous assessment of antinuclear antibodies 'in toto' (ANA). This assay was based upon the use as substrate of smears of peripheral blood derived from rats infected with Trypanosoma lewisi. T. lewisi possesses...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90192-2
更新日期:1982-01-01 00:00:00
abstract::We describe an ex vivo rat infection model to study protective immunity against Fasciola hepatica at the gut level. An exact number of newly excysted juveniles (NEJs) was injected into a gut segment with an intact blood supply and which was still attached to a live anaesthetized rat. NEJs that penetrated the gut wall ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(98)00026-x
更新日期:1998-04-15 00:00:00
abstract::A method is described for isolation of antibody-forming cells with the help of cluster formation. This was achieved by incubating spleen cells from immunized mice with sheep erythrocytes at 37 degrees C. During incubation rabbit-anti-mouse immunoglobulin serum was present in order to facilitate the formation of cluste...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90103-4
更新日期:1976-01-01 00:00:00
abstract::Polymorphisms of immune defence genes may act as disease modifiers and are studied by many researchers. A conclusive analysis of the impact of genetic variations typically requires a large number of sample specimens, and in retrospective studies this may include samples of reduced quality, e.g. formalin-fixed paraffin...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2007.05.006
更新日期:2007-07-31 00:00:00
abstract::Protein tyrosine kinase Syk is known to play critical roles in the signal transduction from receptors for Fc portion of immunoglobulins (FcRs) and B cell receptor complex (BCR). Its importance was well studied in Fc epsilon RI-induced activation of mast cells; therefore, Syk inhibitors are expected to have anti-allerg...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(02)00188-6
更新日期:2002-10-15 00:00:00
abstract::We have investigated the correlation between results obtained by three different methods (semi-quantitative RT-PCR, ELISA and ELISPOT) used to measure cytokine expression by mouse leukocytes. The production of the cytokines tumour necrosis factor-alpha (TNF alpha), interferon-gamma (IFN gamma) and interleukin-4 (IL-4)...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00033-1
更新日期:1997-05-12 00:00:00
abstract::Although many different protocols for transfection of lymphoid cell lines exist, successful DNA transfer into primary B lymphocytes has, to date, not been demonstrated. We now describe a simple method for gene transfer into highly purified normal and malignant B lymphocytes by electroporation. Using a powerful express...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90321-l
更新日期:1990-10-04 00:00:00
abstract::A new method is described for demonstrating the individual subclasses of human IgG immunoglobulins by means of analytical isotachophoresis. Individual subclasses of IgG purified from sera of multiple myeloma patients have distinct mobility patterns. When a mixture with purified IgG for each of the 4 subclasses is anal...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90163-7
更新日期:1979-01-01 00:00:00
abstract::A sensitive assay for the simultaneous detection of multiple serum antibodies by flow cytometry was developed. Polystyrene microspheres of 5, 7 and 9.3 micron in diameter were used as solid supports for the attachment of three different antigen preparations from Candida albicans. These antigens were a whole cell extra...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90206-8
更新日期:1989-01-17 00:00:00
abstract::Etanercept, a recombinant human tumor necrosis factor (TNF) receptor Fc fusion protein is an effective treatment option in adults with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis or plaque psoriasis and paediatrics with juvenile idiotypic arthritis and plaque psoriasis. Patent expiration in Europ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.03.007
更新日期:2017-08-01 00:00:00
abstract::CD3-specific antibodies have shown clinical efficacy in both transplantation and autoimmunity. However, targeting CD3 in this way can lead to T-cell activation and a serious cytokine release syndrome mediated by Fcγ receptor binding. An in vivo mouse model has been developed using severe combined immunodeficient (SCID...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.07.001
更新日期:2012-10-31 00:00:00
abstract::Laurell crossed immunoelectrophoresis (two-dimensional electroimmunodiffusion) was used to prepare minute amounts of purified parasite antigens complexed with their precipitating antibodies obtained from rabbits. These complexes, emuslified in Freund's complete adjuvant, were then used to prime rabbits for selective p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90273-9
更新日期:1978-01-01 00:00:00