Abstract：
:A novel enzyme-linked bio-immunoassay (bio-ELISA) has been developed to detect interferon-gamma (IFN-gamma) induced HLA-DR antigen on the surface of human tumor cells. Cells are cultured at 37 degrees C in 96-well microtiter plates in the presence of IFN-gamma for 2 days. After fixation with reagent alcohol the HLA-DR antigen is detected using a monoclonal antibody, followed by goat anti-mouse IgG-HRP conjugate. Four human cell lines were evaluated and all expressed HLA-DR in response to IFN-gamma in a dose-related fashion. Based on sensitivity, reproducibility and absence of antiproliferative effect by IFN-gamma, the COLO 205 cells (colon adenocarcinoma) were determined to be optimal. The bioassay is sensitive to 0.3 ng/ml IFN-gamma with a range to 10 ng/ml. The specificity of HLA-DR induction by IFN-gamma was demonstrated using an isotype specific monoclonal antibody as well as IFN-gamma neutralizing monoclonal and polyclonal antibodies. The effect of other cytokines on HLA-DR induction with COLO 205 cells was also investigated in this bioassay and only IFN-beta and interleukin-1 (IL-1) showed slight induction of HLA-DR. IFN-alpha had no effect at the concentration tested. Evaluation of assay parameters including reproducibility, sensitivity, simplicity, speed, cost and ability to standardize support the conclusion that this bioassay is a substantial improvement over the routinely used viral inhibition assay as a measure of IFN-gamma biological activity. The bio-ELISA technique also has potential applications for the quantitation of other cellular surface antigens induced by cytokines.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Gibson UE,Kramer SMdoi
10.1016/0022-1759(89)90083-5subject
Has Abstractpub_date
1989-12-20 00:00:00pages
105-13issue
1-2eissn
0022-1759issn
1872-7905pii
0022-1759(89)90083-5journal_volume
125pub_type
杂志文章abstract：:We have designed a cytometry-based competition assay to evaluate peptide binding to empty recombinant HLA class II molecules. The efficiency of this assay was evaluated using recombinant HLA-DP0401 molecules (HLA-DP) produced in insect cells and 13 peptides from human telomerase reverse transcriptase (hTERT). We demon...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.018
更新日期：2011-08-31 00:00:00
abstract：:A simple method for the simultaneous isolation of IgG2, IgG1, IgA and IgM from porcine colostral whey is described. It makes use of DEAE-cellulose chromatography followed by repeated Sepharose 6B exclusion chromatography after the removal of fat, casein and lipoproteins, Fused rocket immunoelectrophoresis is used to s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90058-8
更新日期：1977-01-01 00:00:00
abstract：:A leishmania amastigote radiobinding assay has been developed using organisms labeled with tritiated uracil. These labeled amastigotes resemble freshly isolated unlabeled amastigotes in metabolic activity, buoyant density, morphology, viability and their ability to transform into promastigotes. Organisms routinely inc...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90053-x
更新日期：1990-07-03 00:00:00
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
更新日期：1987-06-26 00:00:00
abstract：:Ferromagnetic iron dextran particles were prepared by reacting a mixture of ferrous chloride and ferric chloride with dextran polymers under alkaline conditions. Particles purified by gel filtration chromatography were in the size range of 30-40 nm, had an electron dense core of about 15 nm, were stable against aggreg...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90007-2
更新日期：1982-08-13 00:00:00
abstract：:We evaluated an enzyme-linked immunosorbent assay (ELISA) for antibodies to native DNA (nDNA) in which protamine was used to link DNA to polystyrene. Elevated anti-nDNA was largely restricted to patients with systemic lupus erythematosus (SLE), and within this group good correlation between ELISA and the ammonium sulf...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(83)80009-x
更新日期：1983-10-28 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：:Radioimmunoassays are usually developed empirically since there have been few established rules which cover general RIA behaviour. Evidence is presented which enables the delineation of 2 types of RIA with distinct group characteristics. (1) Simple RIA, conforming to the law of mass-action kinetics, is a result of uni...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90441-7
更新日期：1985-11-28 00:00:00
abstract：:Monoclonal antibodies (Mab) are the fastest growing group of biopharmaceuticals in development. For production in mammalian cells, the four polypeptide chains of the immunoglobulin diheterotetramer must be assembled prior to exit from the endoplasmic reticulum. Various recombinant Mab expression vectors have been deve...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.10.010
更新日期：2007-01-10 00:00:00
abstract：:The goals for the development of multiple antigen peptides (MAP) are to provide a rational and unambiguous system to multimerize different types of synthetic peptide antigens and to attach immunomodulating molecules for targeting and delivery. These goals have been largely realized and new designs of MAPs now permit a...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(96)00066-x
更新日期：1996-09-13 00:00:00
abstract：:Rapid development in design and production of recombinant antibodies and antibody fragments specific for cell surface markers opens new opportunities for targeted delivery of therapeutic or imaging agents. However, the progress in this field is slowed by inactivation of many antibodies by chemical conjugation of paylo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.03.011
更新日期：2004-06-01 00:00:00
abstract：:The migration inhibition by soluble PPD of peripheral blood mononuclear cells showed a much better reflection of delayed hypersentitivity to tuberculin in man than the migration inhibition of buffy coat cells. The migration index (MI) with mononuclear cells in a group of tuberculin positive subjects was 0.51 +/- 0.14 ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90012-0
更新日期：1976-01-01 00:00:00
abstract：:We developed a reliable method for reconstituting thymectomized, lethally irradiated guinea pigs. Injection of 2.5-10 X 10(7) syngeneic bone marrow cells into adult thymectomized, lethally irradiated guinea pigs produced survival of 46-100% of treated animals. Gentamycin sulfate (5 mg/kg of body weight) for 10 days wa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90187-x
更新日期：1984-05-25 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：:Establishment of humoral immunity against pathogens is dependent on events that occur in the germinal center and the subsequent induction of high-affinity neutralizing antibodies. Quantitative assays that allow monitoring of affinity maturation and duration of antibody responses can provide useful information regardin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.11.026
更新日期：2014-02-01 00:00:00
abstract：:A sensitive method for staining proteins after transfer from polyacrylamide gels to nitrocellulose paper is described. Transferred proteins are first derivatized by reaction of the nitrocellulose replica with sulfosuccinimidobiotin and are then reacted sequentially with streptavidin, rabbit anti-streptavidin, and hors...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90504-1
更新日期：1986-08-21 00:00:00
abstract：:Rhesus monkey serum was applied to a protein A-Sepharose column at pH 7.3. IgG was not detectable in the unbound effluent. Bound protein was eluted with a pH gradient of citrate-phosphate buffer. Two major elution peaks reproducibly resulted at pH 4.85-4.9 and pH 4.35-4.43, respectively, with only slight variability a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90170-3
更新日期：1982-01-01 00:00:00
abstract：:The measurement of the Epstein-Barr viral load in peripheral blood has been recognised as an important way of monitoring the response to treatment in patients with Epstein-Barr virus (EBV)-related malignancies. In particular, EBV load in transplant recipients can be used as a predictive parameter for Post-transplant L...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00333-2
更新日期：2002-12-15 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：:Fab's with hinges based on the human gamma1 sequence containing 1, 2, or 4 cysteines have been produced by high level Escherichia coli periplasmic secretion, and coupled in vitro by reduction/oxidation to form F(ab')2. We find that the F(ab')2 made with hinges containing 2 or 4 cysteines have a high level (approximate...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00061-1
更新日期：1998-08-01 00:00:00
abstract：:The in vitro assessment of T-cell-mediated cytotoxicity plays an important and increasingly relevant role both in preclinical target evaluation and during immunomonitoring to accompany clinical trials employing targeted immunotherapies. For a long time, the gold standard for this purpose has been the chromium release ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2014.01.012
更新日期：2014-03-01 00:00:00
abstract：:Among the many methods currently available for quantifying mRNA transcript abundance, reverse transcription-polymerase chain reaction (RT-PCR) has proved to be the most sensitive. Recently, several protocols for real-time relative RT-PCR using the reporter dye SYBR Green I have appeared in the literature. In these met...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00103-0
更新日期：2003-12-01 00:00:00
abstract：:A new, relatively simple, spectrophotometric technique has been developed which is useful for accurately determining the extent of chromophore labeling of proteins. Often the absorbance spectra and extinction coefficients of dye/protein conjugates are strongly affected by changes in the chromophore microenvironment th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90051-g
更新日期：1991-10-25 00:00:00
abstract：:The automated complement fixation system presented in this paper differs from those described by Studievic et al. (1971) and Vargues and Henley (1974). In the one developed here only one sampler is used with a wheel presenting a double row of cups. This system can assay 60 samples per h and can be modified to double t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90183-6
更新日期：1976-01-01 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：:The influence of size heterogeneity of IgA on its immunonephelometric (IN) assay was studied using highly purified preparations of monoclonal (MC) or polyclonal (PC) monomeric (M), dimeric (D), trimeric (T) and tetrameric (Q) IgA, as well as of secretory IgA (sIgA). Concentrations measured by optical density (OD) were...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90381-7
更新日期：1982-01-01 00:00:00
abstract：:Recent data generated in our laboratory indicates that human lymphokine-activated killer (LAK) cells can be induced in several commercially available serum-free medium formulations. Interestingly, LAK cells were also generated in RPMI 1640 medium plus 1000 U/ml IL-2 without addition of human serum. However, higher lev...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90239-6
更新日期：1988-05-09 00:00:00
abstract：:The aim of this study was to compare the techniques of indirect immunofluorescence assay (IFA) and flow cytometry to clinical and laboratorial evaluation of patients before and after clinical cure and to evaluate the applicability of flow cytometry in post-therapeutic monitoring of patients with American tegumentary l...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.11.002
更新日期：2013-01-31 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：:We have developed a standardized procedure for the isolation of monocytes from peripheral blood by negative selection using magnetic polymer particles coated with monoclonal antibodies against T and B lymphocytes. The average purity of the monocyte suspension was 85%, and monocyte recovery was 72% from Ficoll-Hypaque ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90397-x
更新日期：1991-03-01 00:00:00
