Abstract:
:Fibroblasts are capable of secreting a number of inflammatory mediators and cytokines and may exhibit a marked heterogeneity in this capacity. The relative frequency of cytokine-secreting fibroblasts in chronic inflammatory connective tissue disorders may affect the amount of these molecules secreted locally and dictate the intensity or chronicity of the disease process. We have devised a simple, in situ immunodetection method for assessment of the frequency of actively secreting cells in adherent cell cultures. Our technique is based on the principle of immunoprinting and is coupled with an enzyme-linked immunodetection system. The methodology, sensitivity, specificity and accuracy of this technique are described.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Dongari-Bagtzoglou AI,Ebersole JLdoi
10.1016/s0022-1759(96)00147-0subject
Has Abstractpub_date
1996-11-13 00:00:00pages
145-54issue
2eissn
0022-1759issn
1872-7905pii
S0022175996001470journal_volume
198pub_type
杂志文章abstract::Phagocyte functions such as chemotaxis and phagocytosis involve the rapid and transient development of cellular polarity. Study of this highly complex spatial and temporal cellular remodelling has been limited by the static nature of immunofluorescence and immunogold microscopy and because biochemical techniques are n...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/s0022-1759(99)00155-6
更新日期:1999-12-17 00:00:00
abstract::Mast cells are important effector cells of allergy and are involved in the pathology of many other diseases. Measurement of β-hexosaminidase activity, the most commonly used method for evaluation of murine mast cell activity, requires a large number of cells and thus is of limited utility for studying mast cells in mo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.02.014
更新日期:2012-05-31 00:00:00
abstract::Melanins are notoriously difficult to work with because of their unique physical and chemical properties. The study of melanins is hampered by the scarcity of melanin-specific reagents and serological techniques. In this study we describe modifications to the standard method for the isolation of melanins from in vitro...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00255-6
更新日期:2000-10-20 00:00:00
abstract::Historically, tumour markers are substances selectively released by tumour cells in the blood stream so that they can be detected in the serum or other body fluids for clinical monitoring of various malignancies. The term is now extended to cell or tissue characteristics, such as cytogenetic markers, oncogenes or abno...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(92)90072-2
更新日期:1992-06-24 00:00:00
abstract::The method of extraction of outer membrane proteins (OMPs), the conditions of electrophoretic transfer, and the conditions of antibody binding, were compared in Western blotting studies of Neisseria meningitidis outer membrane antigens. The OMP profiles obtained by SDS-PAGE of outer membrane vesicles extracted with li...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90383-7
更新日期:1990-12-05 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::We describe here a lateral flow-based assay (LFA) for the detection of antibodies against immunodominant antigen Pgp3 from Chlamydia trachomatis, the causative agent of urogenital chlamydia infection and ocular trachoma. Optimal signal detection was achieved when the gold-conjugate and test line contained Pgp3, creati...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2016.05.008
更新日期:2016-08-01 00:00:00
abstract::A serum-free lymphocyte culture medium is described in which serum is replaced by bovine serum albumin, transferrin, insulin, ethanolamine and a mixture of saturated and unsaturated fatty acids (linoleic acid, oleic acid and palmitic acid). In this serum-free medium proliferative and cytotoxic responses induced in mix...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90450-2
更新日期:1984-08-03 00:00:00
abstract::The performance of immunoassays for the detection of autoantibodies is of critical importance to the diagnosis and assessment of patients with systemic lupus erythematosus (SLE). Our objective was to compare 3 multiplexed assays for measurement of multiple autoantibodies and their association with global disease activ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.04.005
更新日期:2010-06-30 00:00:00
abstract::Transduction of bone marrow stem cells with retroviral expression vectors represents a cheaper and more rapid alternative to conventional transgenesis for studies of in vivo gene function. However, achieving tissue-specific expression of genes inserted into retroviral vectors is notoriously difficult. We have develope...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.06.007
更新日期:2010-08-31 00:00:00
abstract::In vitro cell culture models can be of great value in order to further analyze the regulatory mechanisms underlying the inappropriate function of the immune system in diseases such as autoimmunity and cancer. Cell culture conditions have to be well controlled in a way that they mirror the in vivo situation. The object...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2003.07.013
更新日期:2003-11-01 00:00:00
abstract::Several methods have been used to measure anti-double-stranded DNA auto-antibody (anti-dsDNA). Our aim was to determine the most efficient strategy to test anti-dsDNA in systemic lupus erythematosus (SLE). In this study, anti-dsDNA and anti-nuclear antibody (ANA) tests were requested for 644 patients. Anti-dsDNA was t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.09.003
更新日期:2015-12-01 00:00:00
abstract::The concept of TH17 stemness is attracting increasing attention in the field of tumor immunology. The expression of stem cell-like properties and the promotion of long-term immunity by TH17 cells are also of outmost relevance for autoimmunity. Studying two mouse models of multiple sclerosis (MS), we show that CNS anti...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2013.09.010
更新日期:2013-12-15 00:00:00
abstract::Bromodeoxyuridine (BrdU) is used to label synthesizing DNA and to chase label-retaining cell (LRC). As stem cells divide slowly in adult tissues, they can be visualized as LRCs. In order to identify LRCs in hard tissue, we examined optimal conditions of fixation, demineralization, and DNA denaturation/antigen retrieva...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.07.013
更新日期:2008-11-30 00:00:00
abstract::This work presents a method for analyzing protein microarrays using a colorimetric nanogold probe coupled with silver enhancement (gold-silver detection). In this method, the gold nanoparticles were introduced to the microarray by the specific binding of the gold-conjugated antibodies or streptavidins and then coupled...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2003.11.008
更新日期:2004-02-15 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::In inbred or outbred mice lysis of virally modified cells by thymus-derived lymphocytes occurs only within a H-2 compatible system. Peritoneal macrophages provide a convenient source of syngeneic target cells. They are readily infected in vitro with LCM virus, and show high levels of specific 51Cr release when exposed...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(75)90120-9
更新日期:1975-09-01 00:00:00
abstract::Human peripheral blood lymphocytes were stained simultaneously with the lipophilic membrane label PKH-2 and surface markers and analysed by multiparameter flow cytometry. A biphasic staining distribution was observed for T (CD3+) as well as B (CD19+) cells. Within the T subset, CD4+ cells stained almost exclusively wi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(92)90211-b
更新日期:1992-09-18 00:00:00
abstract::Nitrocellulose was activated with divinyl sulfone, a spacer of ethylenediamine, and glutaraldehyde. The aldehyde groups on the activated nitrocellulose, Nit-CHO, were stable through one month at 4 degrees C. Peptides were attached to the membrane by reaction of the amino group with the free carbonyl, forming peptide b...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90197-4
更新日期:1990-08-07 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 periodate oxidation and chromium chloride coupling methods were compared for their ability to sensitize indicator erythrocytes with staphylococcal lipoteichoic acid (LTA) for the detection of specific antibody. Erythrocytes, sensitized with periodiate-activated lipoteichoic acid, were found to be superior for use ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90070-8
更新日期:1980-01-01 00:00:00
abstract::Colloidal gold particles coated with rabbit anti-HCG (Au-(anti-HCG)) were used in a spectrophotometric 'sandwich' SPIA for HCG. The effects of several reagent properties and assay variables were investigated in order to optimize the assay. Best results were obtained with 55 nm particles. Optimum assay conditions invol...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90244-2
更新日期:1983-08-26 00:00:00
abstract::A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modifica...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90332-6
更新日期:1984-02-10 00:00:00
abstract::We developed a simple and sensitive enzyme-linked immunosorbent assay (ELISA) for human pulmonary surfactant protein D (SP-D). Human SP-D was purified from bronchoalveolar lavage fluids of patients with pulmonary alveolar proteinosis. Nine monoclonal antibodies (MAbs) were established from BALB/c mice immunized with t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)90295-x
更新日期:1994-08-01 00:00:00
abstract::We describe here a spot test to detect antibodies to non-erythroid cell surface antigens, using ethidium bromide staining to visualize areas of target cell lysis. This assay allows rapid screening of large numbers of samples of culture supernatants, antisera, or ascitic fluids. Antibody titers determined by this spot ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90367-2
更新日期:1982-01-01 00:00:00
abstract::We found that some plasma proteins such as haptoglobin (Mr 200,000) and alpha 1-acid glycoprotein (Mr 40,000) do not bind at all to nitrocellulose membranes in classical transfer conditions. In contrast, these proteins can be clearly identified when a charged nylon membrane is disposed on the anodal side of the nitroc...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90226-8
更新日期:1988-03-16 00:00:00
abstract::The production and characterisation of 17 monoclonal antibodies to human parathyroid hormone-related protein (PTH-rP) 1-34 is described. Five of the antibodies were shown to be of high avidity (Ka 4 X 10(10)-1.9 X 10(11) L/M) and able to detect 15-100 pg PTH-rP 1-34 per tube by RIA. None cross-reacted with PTH 1-34, a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90346-w
更新日期:1990-02-20 00:00:00
abstract::AMPKalpha is a subunit of AMP-activated protein kinase (AMPK), a heterotrimeric enzyme that works as a fuel sensor activated in response to the depletion of cellular ATP. AMPKalpha is considered as a master switch in regulating glucose and lipid metabolism. Determining its presence in patient sera may help in diagnosi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2009.10.001
更新日期:2009-12-31 00:00:00
abstract::Contamination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis samples has commonly produced artefactual bands when sensitive detection systems, such as silver staining or immunoblotting procedures, have been used. Evidence suggests that these bands originate from human skin contamination of reagents and ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90090-7
更新日期:1988-05-25 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