Abstract：
:Cytoplasmic free calcium [( Ca2+]i) is a key intracellular messenger in many cell types. We have used fluo-3, a recently developed calcium probe, to study [Ca2+]i in resting and stimulated human peripheral blood T lymphocytes. The spectral properties of fluo-3 permit analysis of [Ca2+]i in flow cytometers with a 488 nm argon laser excitation source and fluorescein filter settings. The data obtained with fluo-3 are both qualitatively and quantitatively in good agreement with the data in the literature. After stimulation of T lymphocytes with the mitogens phytohaemagglutinin, concanavalin A and with OKT3, and anti-CD3 monoclonal antibody, a biphasic [Ca2+]i response was observed, with an early EGTA-insensitive [Ca2+]i rise, followed by an EGTA-sensitive sustained [Ca2+]i plateau. Non-mitogenic monoclonal antibodies directed against the CD5, CD28 and CD7 T cell surface antigens elicited [Ca2+]i rises only when crosslinked on the cell surface with goat anti-mouse IgG. Conversion of fluorescence data into absolute [Ca2+]i values by means of a non-disruptive calibration procedure, yielded a [Ca2+]i of 107 +/- 18 nM (mean +/- SD, n = 13) in resting T lymphocytes. Time-dependent loss of cellular dye content limits the precision of the calibration procedure in experiments of longer duration. We conclude that fluo-3 promisingly extends the potential application field of flow cytometers with 488 nm argon lasers to [Ca2+]i studies in T lymphocytes.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Vandenberghe PA,Ceuppens JLdoi
10.1016/0022-1759(90)90069-8subject
Has Abstractpub_date
1990-03-09 00:00:00pages
197-205issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(90)90069-8journal_volume
127pub_type
杂志文章abstract：:The effect of various physical or chemical treatments of splenic stimulator cells on their endogenous, mitogen-inducible IL-2 production and on their ability to induce IL-2 production in clonally developing helper T lymphocytes was investigated. While most methods (T cell depletion by monoclonal antibodies plus comple...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90241-4
更新日期：1988-05-09 00:00:00
abstract：:Ammonium-chloride-containing solutions (AC) are routinely used to lyse red blood cells during preparation of PBMC. Although exposure to AC has been described to affect the ultrastructural appearance of large granular lymphocytes and to temporarily inhibit cytolytic activity of PBMC preparations, the cellular basis of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00326-x
更新日期：2001-06-01 00:00:00
abstract：:The existing methods for phagocytosis evaluation are inadequate for assessing all the real events occurring during phagocytosis, or for continuously following the kinetics of the process. Our purpose is to establish the use of luminous bacteria as an object for phagocytosis. The bioluminescence test offers an easy and...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90443-x
更新日期：1983-11-25 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：:An ELISA method for the detection of monoclonal antibodies (MAb) to the T3-T cell antigen receptor (TCR) complex was devised. The T3-TCR complex was solubilised using digitonin and a rat anti-T3 MAb (Campath 3) was used to bind it to an ELISA plate. Normal rat serum was used to block cross-reactivity between the rat M...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90280-3
更新日期：1988-01-21 00:00:00
abstract：:Helper and suppressor activity of T cell lines were investigated using an assay system which is based on measurement by the enzyme-linked immunosorbent assay (ELISA) of the antibody produced in vitro. T cell lines were established from spleens of BALB/c mice immunized with ovalbumin and human serum albumin. The T cell...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90063-7
更新日期：1983-02-25 00:00:00
abstract：:One of the major endpoints of cellular immunological tests remains the quantitative assessment of lymphocyte proliferation as estimated from incorporation of tritiated thymidine into dividing cells. These data are usually generated by beta scintillation counting as strings of counts per minute which require considerab...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90163-a
更新日期：1991-04-25 00:00:00
abstract：:The majority of immunological processes are mediated by cell-to-cell contact or receptor-ligand interactions that transmit intracellular signals and affect the regulation of transcription in the nucleus. As a consequence, precursor cells develop into their respective lineages and cells differentiate further during an ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(01)00304-0
更新日期：2001-04-01 00:00:00
abstract：:A mixed hemadsorption (MH) test for naturally acquired rubella immunity and immune responses after rubella vaccination has been compared with the hemagglutination-inhibition (HI) test, the immunodiffusion (ID) test, the neutralization test (NT), the hemolysis-in-gel (HIG) test and the indirect immunofluorescence (IF) ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90282-3
更新日期：1980-01-01 00:00:00
abstract：:We have developed a family of monoclonal antibodies directed against the retinoblastoma gene product (p110RB). One of these monoclonal antibodies, 3C8, binds p110RB near the C-terminal end of the protein (aa886-aa905). It was characterized by immunoblotting, ELISA, fluorescence-activated flow cytometry and immunohisto...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90267-4
更新日期：1994-03-10 00:00:00
abstract：:Levels of acute phase cytokines secreted ex vivo by peripheral blood mononuclear cells (PBMCs) have been shown to be associated with clinical conditions or histologic lesions in renal transplant recipients. One of the limiting factors for the potential use of this assay as a diagnostic tool is the incubation time need...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.01.003
更新日期：2011-03-07 00:00:00
abstract：:For microcytotoxicity, when numbers of effector or target cells are limited, the method of Takasugi and Klein (1970) is at present the most convenient way of assessing cytotoxicity. In this procedure, target cell survival is affected by minor variations in culture conditions or washing vigour and the variations may be...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90384-2
更新日期：1982-01-01 00:00:00
abstract：:Monoclonal antibodies (mAbs) have emerged as a promising tool for cancer therapy. Differing approaches utilize mAbs to either deliver a drug to the tumor cells or to modulate the host's immune system to mediate tumor kill. The rate by which a therapeutic antibody is being internalized by tumor cells is a decisive feat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.04.024
更新日期：2015-08-01 00:00:00
abstract：:A recombinant retroviral vector with an inserted bacterial neomycin resistance (neo) gene was used to transfer in vitro neomycin resistance (neoR) into murine cytotoxic lymphocyte precursors (CLP). The infection protocol involved co-cultivation of mitogen-activated splenic T-blasts with irradiated cells that produced ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90036-0
更新日期：1986-05-01 00:00:00
abstract：:Monoclonal antibodies (MAbs) were selected for specific binding to spent media of cultured tumor cells. Out of more than 12,000 hybridomas screened, 19 were selected in preliminary inhibition assays for secretion of MAbs which detected antigens in cancer patients' sera. Antibody CO 29.11, which was studied in detail, ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90169-3
更新日期：1985-06-12 00:00:00
abstract：:Small haptens such as methylamphetamine (MW 149) cannot, on their own, induce an immune response. It is also unlikely that they fill the binding site of any antibody that recognises them. Under such circumstances any attached label might be expected to enter the area of the binding site and exert an influence on overa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90375-p
更新日期：1991-07-05 00:00:00
abstract：:A radioimmunoassay is described that allows rapid determination of a urinary oligosaccharide -- Glc alpha 1-6Glc alpha 1-4Glc alpha 1-4Glc [(Glc)4] -- at concentrations greater than 2 pmol/microliter. Antibodies produced in rabbits immunized with the phenethylamine derivative of (Glc)4 coupled to keyhole limpet hemocy...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90215-0
更新日期：1982-01-01 00:00:00
abstract：:Lymphocytes preincubated with various activities of Na251CrO4 were tested for their ability to induce a local, popliteal lymph node graft-versus-host (GVH) reaction. Incubation with 100 micronCi of the isotope per ml of cell suspension impaired the GVH activity, 25 micronCi gave a reduced response in 1 out of 3 experi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90153-9
更新日期：1978-01-01 00:00:00
abstract：:Many research and clinical applications require large quantities of full-length antibodies with long circulating half-lives, and production of these complex multi-subunit proteins has in the past been restricted to eukaryotic hosts. In this report, we demonstrate that efficient secretion of heavy and light chains in a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00036-4
更新日期：2002-05-01 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：:A conjugate of 2-aminohexylamino-4-ethylamino-6-isopropylamino-1,3,5-triazine (AHA), a derivative of the herbicide atrazine, with dextran as carrier has been synthesized and used as the coating antigen in ELISA procedures. The quantification of terbutryn, atrazine and prometryn in ELISA formats using monoclonal antibo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90100-8
更新日期：1992-07-06 00:00:00
abstract：:Pertussis toxin is known to elicit lymphocytosis in whooping cough patients and experimental animals, by blocking the extravasation of lymphocytes and stimulating their release from lymphoid organs such as the thymus. The mechanisms responsible for these unique effects of PT are not fully understood. The effect of per...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90090-3
更新日期：1991-11-22 00:00:00
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
更新日期：1983-10-28 00:00:00
abstract：:A dot-ELISA technique for the detection of G. lamblia specific antigen in stool eluates of clinical cases of giardiasis was developed and evaluated employing monospecific antibodies to a G. lamblia specific coproantigen with a molecular mass of 66 kDa. The assay detected 22 (91.7%) of the 24 microscopically confirmed ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90030-j
更新日期：1991-03-21 00:00:00
abstract：:5-Bromo-2-deoxyuridine (BrdUrd) is a thymidine analogue which can be used to detect DNA synthesis in cells. The present paper describes a method for the fixation and DNA denaturation of cells that affects neither the morphology of the cells not their adhesion to a glass surface. The cells were fixed in absolute ethano...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90401-2
更新日期：1988-04-06 00:00:00
abstract：:ELISAs for urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) have shown that tumor levels of these molecules are prognostic parameters in breast cancer as well as other types of cancer. These ELISAs measure the total amount of the given component, including preforms, active, inac...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00008-2
更新日期：1997-04-11 00:00:00
abstract：:The fluorescence activated cell sorter (FACS) was used with an indirect membrane immunofluorescence technique to detect antibody against the Forssman antigen, a glycosphingolipid. Sheep erythrocytes, which contain Forssman antigen as a major membrane glycosphingolipid, were used as the target antigen. Detection of the...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90098-4
更新日期：1983-02-25 00:00:00
abstract：:alpha-2-macroglobulin (alpha-2-M), a plasma proteinase inhibitor, plays an important role in the pathogenesis of lung inflammation. The purpose of this study was to develop a sensitive ELISA assay for rabbit alpha-2-M to allow us to define the role of this protein in a rabbit model of lung inflammation/injury. Therefo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00278-8
更新日期：2002-12-15 00:00:00
abstract：:Irradiated human peripheral blood lymphoid cells undergo apoptosis and progressively exhibit typical changes in light scatter and plasma membrane integrity that can be easily tracked by flow cytometry. Using this model, we assessed the capacity of a newly developed fluorochrome, Apostain, in identifying early apoptosi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00054-9
更新日期：1997-06-23 00:00:00
abstract：:Elucidation of local immune response at the cervix is important for understanding and evaluating STD vaccine approaches currently being proposed. However, no well-validated method exists for the collection of cervical secretions for evaluation of cervical immune response. The purpose of this study was to determine the...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00037-x
更新日期：1999-05-27 00:00:00
