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 surprisingly, we have observed that 2ME induces the conversion of CD138(-) B lymphocytes into CD138(+) cells of phenotype similar to immunoglobulin (Ig)-secreting plasma cells. Normal human B lymphocytes expressing CD138 increased in response to 2ME in a dose-dependent fashion, from 2% at baseline up to 31% in cells cultured in the presence of 0.75 microM 2ME. Moreover, most of the converted cells were also CD27(+) and secreted high levels of IgG (151 microg/10(6)cells/24h). IEF studies revealed that conversion occurred in a polyclonal manner. We then exploited this effect of 2ME to gain further insights into the molecular mechanisms that govern changes in transcription factors involved in plasma cells differentiation. Plasma cells generated by 2ME treatment of normal human B lymphocytes expressed elevated levels of IRF4 and reduced levels of Pax5 and Bcl-6. Similarly, levels of XBP-1 and Blimp-1 transcripts were increased. Our results suggest that the differentiation of peripheral blood B lymphocytes into plasma cells requires a similar modulation of transcription factors expression that for tonsil and bone marrow B lymphocytes.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Cayer MP,Drouin M,Proulx M,Jung Ddoi
10.1016/j.jim.2010.02.012subject
Has Abstractpub_date
2010-04-15 00:00:00pages
29-39issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(10)00046-3journal_volume
355pub_type
杂志文章abstract：:This study was designed to assess the use of flow cytometry to observe the binding, under physiological conditions, of anti-lipopolysaccharide (LPS) monoclonal antibodies (mAbs) to whole bacteria, and to compare this with the more conventional whole cell ELISA and immunoblotting techniques. The bacteria consisted of t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90363-z
更新日期：1990-10-19 00:00:00
abstract：:A new strategy has been developed for enzyme immunoassay of small molecules. The method is based on the formation and estimation of the complex Ab.H-H.Ab* where Ab and Ab* are immobilised and labeled antibodies respectively and H-H is a synthetic bis-analogue of the hapten H. The amount of the complex formed is invers...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(12)80005-6
更新日期：1992-03-04 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90268-9
更新日期：1980-01-01 00:00:00
abstract：:A sensitive direct assay has been developed to quantitate antigen specific helper factors (ThF). The assay depends on the ability of ThF to block the binding of free antibody to immobilized antigen. It has been used to follow the course of purification of ThF produced in short-term cultures of in vitro primed helper c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90098-x
更新日期：1984-02-24 00:00:00
abstract：:We describe the preparation of a new rat monoclonal antibody (CRC1) to the N-terminal sequence of the 43 kDa subunit of human ovarian inhibin, and its use together with other anti-peptide monoclonal antibodies, in two-site immunoassays for the detection of inhibin-related material in biological fluids. The Fab fractio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90342-5
更新日期：1993-10-15 00:00:00
abstract：:Phycobilisomes are cyanobacterial photosynthetic energy transfer complexes partly composed of phycobiliproteins, proteins that are widely used as conjugable fluorochromes for flow cytometry. The brightness and photostability of phycobiliproteins suggest that intact phycobilisomes could constitute even brighter probes ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00367-2
更新日期：2001-08-01 00:00:00
abstract：:A simple technique for raising specific antiserum to the native molecule of Epstein-Barr virus (EBV)-encoded nuclear antigen-2 (EBNA-2) from Raji cells is described. The procedure involves the use of immunoblotting to identify the EBNA-2 polypeptide followed by subsequent excision from an SDS-gel and immunization of e...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90543-0
更新日期：1986-03-13 00:00:00
abstract：:A rapid, direct method for the purification of sheep red cell rosetting lymphocytes (ERFC) was developed. The whole procedure, including rosette formation, density separation and hemolysis could be completed within 10 min. A mixture of human peripheral blood mononuclear cells (PBMC) and 2-aminoethylisothiouronium brom...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90217-7
更新日期：1988-03-16 00:00:00
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
更新日期：1995-10-26 00:00:00
abstract：:Foot-and-mouth disease virus (FMDV) is an important veterinary pathogen which can cause widespread epidemics. Due to the high antigenic variability of FMDV, it is important to undertake mutation analysis under immunological pressure. To study the bovine antibody response at a molecular level, phage display technology ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.01.002
更新日期：2004-03-01 00:00:00
abstract：:A new sandwich immunoassay method for measuring human C5a-des-Arg was developed using monoclonal antibody specifically reactive with C5a-des-Arg. Monoclonal antibodies were obtained from a panel of hybridomas produced by fusion of mouse myeloma cells, P3 X 63-AG8,653, with spleen cells from a CBF1(C57BL/6 X BALB/c) mo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90159-1
更新日期：1987-08-03 00:00:00
abstract：:This communication describes an automated micro-adherence inhibition assay. Tumor-specific immunity was demonstrated with B16 melanoma and MCA-38 colon adenocarcinoma, both of which are syngeneic to the same strain of mouse (C57B16/J). Abrogation of the leukocyte adherence inhibition (LAI) response of sensitized leuko...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90086-2
更新日期：1977-01-01 00:00:00
abstract：:A solid-phase chemiluminescence immunoassay for determining immunoglobulin G (IgG) in eluates of psoriatic scales is described. Polystyrene balls coated with protein A bound the IgG present in the eluates and in human sera. The bound IgG was then detected by a luminol-coupled goat anti-human IgG conjugate, characteriz...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90184-4
更新日期：1984-05-25 00:00:00
abstract：:Investigation of MHC class I endogenous antigen processing and presentation is important for understanding non-self immune recognition, vaccine design, immune evasion by pathogens and tumours, and elucidation of the mechanisms of certain auto-immune diseases. To study the MHC class I antigen-processing pathway, the pr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.02.017
更新日期：2008-06-01 00:00:00
abstract：:We report a procedure for optimisation of Western blotting using protein G-horseradish peroxidase (protein G-HRP) which avoids the false positive reactions often caused by second antibodies and increases the detection of autoantibodies by protein G conjugate. A number of modifications were investigated. Higher concent...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00235-9
更新日期：1997-03-28 00:00:00
abstract：:A novel technique for establishing short term clones of antigen- or mitogen-activated splenic B lymphocytes is described. Spleen cells are plated onto the surface of filter paper discs and subsequently stimulated by antigen or mitogen in situ; activated B cells proliferate and differentiate into pure colonies of cells...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90312-6
更新日期：1985-02-11 00:00:00
abstract：:There is a need for simple and sensitive assays to assess innate and adaptive immune responses to microbial agents and vaccines. Herein, we describe a whole blood method allowing to measure the induction of cytokine synthesis at the mRNA level. The originality of this method consists in the combination of PAXgene tube...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00074-7
更新日期：2003-05-01 00:00:00
abstract：:The ability to isolate antibodies against any antigen of interest has become increasingly important as antibodies have proved their utility both in antigen detection, quantification and as specific in vivo targeting agents. To this end, we have constructed a large antibody phage library in the single chain Fv (scFv) p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.01.014
更新日期：2005-04-01 00:00:00
abstract：:We describe a simple and efficient non-chromatographic method for the purification of murine IgG3 and IgM monoclonal antibodies (MAbs) which takes advantage of their euglobulin properties. Following filtration, ascitic fluid is dialysed against demineralized water and centrifuged at 22,000 X g for 30 min. The resultin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90054-3
更新日期：1988-06-28 00:00:00
abstract：:When limited amounts of antigens are available, it is often difficult to obtain large quantities of polyclonal antisera. Although antisera can be raised in small rodents, yields are usually small. To circumvent these problems, we have designed a modified method for generating polyclonal ascites fluid (AF). Using the a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90108-j
更新日期：1993-09-27 00:00:00
abstract：:The Immune Epitope Database and Analysis Resource (IEDB, http://www.iedb.org) hosts a continuously growing set of immune epitope data curated from the literature, as well as data submitted directly by experimental scientists. In addition, the IEDB hosts a collection of prediction tools for both MHC class I and II rest...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2010.10.010
更新日期：2011-11-30 00:00:00
abstract：:High-titered antisera against human melanoma- and leukemia-associated antigens have been produced in non-human primates. The data from this and from other studies suggest that this model may prove a valuable source of highly specific antisera against a variety of human tumor-associated antigens or alloantigens. Our me...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90048-x
更新日期：1976-01-01 00:00:00
abstract：:An indirect binding assay, the fluorometric immunoassay (FIAX), was established for the detection of anti-M2 antibodies which are specific markers for primary biliary cirrhosis (PBC). Submitochondrial particles (SMP) from beef heart and rat liver and the ATPase-associated antigen (M2) were used. The antigens were fixe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90401-5
更新日期：1983-11-11 00:00:00
abstract：:Cultured human melanoma and gastrointestinal carcinoma cells were detached from substrate and further dissociated by placing the culture vessel into a water-filled ultrasonic cleaner (43 kHz) and sonicating it for 10-50 s. Plating efficiency and long-term growth of three melanoma cell lines were similar after ultrasou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90480-7
更新日期：1987-11-23 00:00:00
abstract：:Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation onl...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(93)90063-d
更新日期：1993-01-04 00:00:00
abstract：:A method is presented for measuring the hemolytic activity of undiluted guinea pig serum. Undiluted serum is placed in contact with a column of sensitized sheep red blood cells in an agar matrix in a capillary tube. After incubation, the length of the zone of lysis is measured. Comparison with a standard curve prepare...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90212-6
更新日期：1979-01-01 00:00:00
abstract：:A micro-assay has been developed for the separate evaluation of phagocytosis and intracellular killing of Staphylococcus aureus by human monocytes and granulocytes. The minimal number of phagocytes required for the investigation of these functional activities of phagocytic cells has been established by performing phag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90095-9
更新日期：1983-02-25 00:00:00
abstract：:A mixed haemadsorption procedure for the detection and localisation of spermatozoal antigens is described. Sperm are immobilised by attaching them to the bases of wells on plastic plates using poly-L-lysine and antisera and sensitised red cells are then added to these wells. The technique uses simple reagents and allo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90242-4
更新日期：1979-05-10 00:00:00
abstract：:The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a mu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.05.005
更新日期：2004-08-01 00:00:00
abstract：:We describe optimal conditions for protein labeling with a new fluorescent probe, 4,7-bis-(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). The labeled proteins are suitable for time-resolved fluorometric applications because BCPDA forms fluorescent complexes with Eu3+ which exhibit very long fluo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90031-2
更新日期：1988-08-09 00:00:00
