Abstract：
:IgA antibodies are key immune effectors against invading pathogens but also possess essential immunoregulatory functions. Detecting and quantifying human IgA+ B-cell subsets and secreted IgA molecules is needed for investigating the protective, modulatory and pathophysiologic roles of IgAs. Here, we produced a recombinant tagged trimeric form of the streptococcal IgA-binding peptide (SAP) by transient transfection-based eukaryotic expression system. The trimeric SAP (tSAP) probe had a higher production yield and apparent binding affinity to human IgA1 and IgA2 immunoglobulins when compared to the dimeric SAP molecule classically used to purify IgAs. tSAP bound both monomeric and dimeric IgAs, and allowed immunoblot detection and ELISA quantification of serum IgA antibodies in humans and non-human primates. Fluorescently labeled tSAP also permitted an accurate quantification of circulating human blood IgA-expressing memory B cells by flow-cytometric analyses. Thus, the easy-to-produce high affinity recombinant tSAP probe we developed is a versatile and valuable tool to quantify secreted and membrane-bound human but also primate IgA immunoglobulins.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Planchais C,Mouquet Hdoi
10.1016/j.jim.2020.112833subject
Has Abstractpub_date
2020-01-01 00:00:00pages
112833eissn
0022-1759issn
1872-7905pii
S0022-1759(20)30117-4journal_volume
484-485pub_type
杂志文章abstract：:The 7th International Workshop on Human Leucocyte Differentiation Antigens (HLDA7) studied a number of newly characterised molecules relevant to human leucocyte differentiation and function. The HLDA organisation, which devised and continues to maintain the CD nomenclature, is responsible, under the auspices of IUIS a...
journal_title：Journal of immunological methods
pub_type：
doi：10.1016/s0022-1759(03)00057-7
更新日期：2003-04-01 00:00:00
abstract：:Efficacious typhoid vaccines for young children will significantly reduce the disease burden in developing world. The Vi polysaccharide based conjugate vaccines (Vi-rEPA) against Salmonella Typhi Vi positive strains has shown high efficacy but may be ineffective against Vi negative S. Typhi. In this study, for the fir...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.07.005
更新日期：2017-11-01 00:00:00
abstract：:The study of organogenesis, tissue-homeostasis and regeneration requires the precise assessment of in vivo cell proliferation. To this end a host of methods have been developed to detect and quantify DNA synthesis in proliferating cells. These include cell labeling with various nucleotide analogues and fluorescence re...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.03.015
更新日期：2018-06-01 00:00:00
abstract：:Certain serum samples produce high background in Western and direct immunoblot assays that detect human serum IgG against specific antigens. We determined that this was due to a reaction between endogenous IgG and the membrane blocking agent (we refer to this as blocking-specific background). Using milk as blocking ag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90259-a
更新日期：1993-01-14 00:00:00
abstract：:Limited data exists regarding antibody avidity for human papillomavirus (HPV). We describe development of a multiplex electrochemiluminescent avidity ELISA for four HPV types (HPV 6, 11, 16, 18) by adding a dissociating step to our established multiplex HPV VLP ELISA. Initial experiments exploring ammonium thiocyanate...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.04.004
更新日期：2017-08-01 00:00:00
abstract：:A new micro-glass-tube leukocyte adherence inhibition (LAI) assay which is appropriate for detecting delayed type hypersensitivity in vitro has been developed for human leukocytes. Enumeration of adherent cells is replaced by a cellular radioimmunoassay determining antibody binding of the monoclonal reagents, OKT4, OK...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90482-5
更新日期：1986-07-24 00:00:00
abstract：:A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90149-8
更新日期：1985-12-27 00:00:00
abstract：:We have developed a solid-phase assay for the quantification of terminal deoxynucleotidyl transferase (TdT) enzymatic activity in crude cellular extracts. Affinity-purified, polyclonal anti-TdT antibodies are bound to the wells of a microtiter plate, and TdT in extracts is then bound to the immobilized antibodies. The...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00004-t
更新日期：1995-04-26 00:00:00
abstract：:An assay to detect circulating immune complexes is described and exemplified with serum from SLE and RA patients. The method is based on selective precipitation of immune complexes with PEG followed by a specific ELISA assay to detect the immunoglobulins present in the precipitate. The immunoglobulins of the precipita...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90186-3
更新日期：1985-02-28 00:00:00
abstract：:Human monoclonal antibodies may be generated by electrofusion of human B lymphocytes with a human/mouse heteromyeloma line. In addition to a fusion protocol optimised for the fusion partners, the activation of B lymphocytes is crucial for fusion and hybrid efficiency. In this study, we initially treated peripheral blo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00431-8
更新日期：2001-09-01 00:00:00
abstract：:Antibody phage display technology is a well established method for selecting specific antibodies against desired targets. Although phage display is the most widely used method of generating synthetic antibodies, it is laborious to perform multiple selections with different antigens simultaneously using conventional ma...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.09.034
更新日期：2011-03-31 00:00:00
abstract：:Immune complexes attached to and ingested by human polymorphonuclear (PMN) cells were quantified by cytofluorometry using a fluorescence quenching assay which permits differentiation between attachment and ingestion. The fluorescence intensity decreased after ingestion as a result of the low pH in the phagolysosomes. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90340-x
更新日期：1983-05-27 00:00:00
abstract：:The aim of the study was to develop a novel system permitting automated analysis of multicolor immunofluorescence-staining of cells in solid tissues which would be comparable to the analytical capacity of flow cytometry. In the user friendly automated data acquisition and image processing system which was established,...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00240-9
更新日期：2000-04-03 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90533-8
更新日期：1986-03-13 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：:This paper describes a modification of the peroxidase technique by which immunophenotyping may be carried out on routinely air-dried blood and bone marrow (BM) smears. The method is simple and quick, requires no special equipment, can be performed on fresh or stored specimens and gives a standard of morphological deta...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90428-x
更新日期：1986-10-23 00:00:00
abstract：:An improved fixation method for antigen detection in lymphoblastoid cells is described. Herpesvirus papio nuclear antigen (HUPNA) could be stained in several transformed lymphoid cell lines by anti-complement immunofluorescence (ACIF). Antibody to HUPNA was detected in many human sera containing antibodies to Epstein-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90162-9
更新日期：1981-01-01 00:00:00
abstract：:We have developed an enzyme-linked immunosorbent assay (ELISA) for the quantitation of C4 nephritic factor (C4NeF). Incubation of the C4NeF-positive serum from patient M.I. with normal human serum (NHS) in the presence of human aggregated IgG (AHG) resulted in the formation of stable C4-C2 complex. No complex was form...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90468-b
更新日期：1990-03-27 00:00:00
abstract：:Serum factors which interact with human peripheral blood lymphocyte Fc gamma receptors (Fc gamma Rs) may be detected in vitro by the EA rosette inhibition assay (EARIA). This assay has been used to detect circulating immune complexes and certain alloantibodies directed against cell surface antigens situated in close p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90222-s
更新日期：1993-02-03 00:00:00
abstract：:Direct reverse transcriptase in situ polymerase chain reaction (RT-in situ PCR) of selected mRNA expression in rat mast cells (MC) and alveolar macrophages (AM) was optimized. Rat peritoneal mast cells (PMC), rat cultured mast cells (RCMC), rat bronchoalveolar lavage cells (BALC) or rat cultured alveolar macrophages (...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00061-7
更新日期：1999-06-24 00:00:00
abstract：:A rapid and highly reproducible protocol which permits the detection of quantities of estrogen receptor as low as 5 fmol/mg protein is described. The separation of free and receptor-bound hormone is achieved by specific immunoprecipitation of the hormone-receptor complex. This procedure can be performed without pertur...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90234-7
更新日期：1991-11-05 00:00:00
abstract：:A high non-specific binding of immunoglobulins to plastic surfaces was noted with a number of rat sera, when tested in an indirect 125I-labeled protein-A assay for detection of cell-surface-bound rat immunoglobulins of various classes and IgG subclasses. This type of non-specific binding was found with all types of Ig...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90243-6
更新日期：1984-01-20 00:00:00
abstract：:2-Methoxyestradiol (2ME), an end-metabolite of 17beta-estradiol, is an antiproliferative agent that is currently being tested in clinical trials for cancer treatment. We hereby report that sub-cytotoxic concentrations of 2ME influence the in vitro proliferation of human peripheral blood B lymphocytes. More surprisingl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.02.012
更新日期：2010-04-15 00:00:00
abstract：:A new and simple method for quantitation of IgM of low molecular weight in sera by radial immunodiffusion in 7% agar gel is described. The test is easy to perform and permits exact determination of low molecular weight IgM concentration down to very small quantities (1 mg%). Results obtained with the method described ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90094-7
更新日期：1978-01-01 00:00:00
abstract：:We have developed specific immunoassays for secretory peroxidase (SP) and for myeloperoxidase (MP) (polymorphonuclear leukocyte-derived peroxidase) in human saliva. Antibodies against SP and MP were produced using bovine milk lactoperoxidase (LP) and human MP as the immunogens, respectively. The methods developed are ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90154-8
更新日期：1991-08-09 00:00:00
abstract：:To reduce the cost, time and waste in screening for HIV and HBV infections a combined assay for HIV-1 and -2 antibodies and HBsAg has been developed. Monoclonal anti-HBs antibodies were co-immobilized with synthetic peptides representing immunodominant regions of HIV-1 and -2. The presence of anti-HIV antibodies in th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90262-6
更新日期：1993-01-14 00:00:00
abstract：:We studied the binding of dog immunoglobulins G, A, M and E to protein A and protein G. Passive cutaneous anaphylaxis (PCA) testing was used for the measurement of dog IgE and enzyme-linked immunosorbent assays (ELISA) were used for the measurements of dog IgG, IgA and IgM. Protein A from lyophilized cells of Staphylo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90335-d
更新日期：1991-12-15 00:00:00
abstract：:An antibody binding-inhibition test is described, which allows the detection of P. falciparum in red blood cells (RBC) infected in vitro, using a crossreacting, monoclonal anti-P. berghei antibody and P. berghei coated microtiter plates. Experiments carried out to determine the coating efficiency of various P. berghei...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90231-5
更新日期：1985-09-03 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：:Use of whole blood for lymphocyte proliferation experiments have advantages over the usual isolated lymphocyte procedure. We have systematically optimized methodology for whole blood lymphocyte proliferation in male Sprague-Dawley rats. Both phytohemagglutinin (PHA) and concanavalin A (ConA) stimulate lymphocyte blast...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00084-n
更新日期：1995-08-18 00:00:00
