Abstract：
:Real-time polymerase chain reaction (PCR) is now becoming an accepted tool for measuring gene expression at the transcriptional level. In this study, a direct comparison between real-time PCR, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunosorbent spot (ELISPOT) assay was performed. When interferon-gamma (IFN-gamma) gene expression was assessed, both ELISA and ELISPOT data strongly correlated to results obtained by real-time PCR. Real-time PCR was subsequently used to measure bovine IFN-gamma (bIFN-gamma) and bovine interleukin-4 (bIL-4) gene expression by antigen stimulated peripheral blood mononuclear cells (PBMC), isolated from bovine herpesvirus-1 (BHV-1) infected animals. BHV-1-infected animals were either non-vaccinated or vaccinated using one of two adjuvants prior to infection. With non-vaccinated infected animals, a Th1 bias occurred, based on IFN-gamma expression exceeding IL-4 expression. The level of cytokine expression, and the IFN-gamma/IL-4 ratio could be significantly affected, depending on the manner in which animals were vaccinated.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Mena A,Ioannou XP,Van Kessel A,Van Drunen Little-Van Den Hurk S,Popowych Y,Babiuk LA,Godson DLdoi
10.1016/s0022-1759(02)00029-7keywords:
subject
Has Abstractpub_date
2002-05-01 00:00:00pages
11-21issue
1-2eissn
0022-1759issn
1872-7905pii
S0022175902000297journal_volume
263pub_type
杂志文章abstract：:The application of freeze-dried paraffin sections for immunohistology eliminates many of the problems associated with the cryostat technique: the sections are thin sufficiently, the cell morphology is improved, there are no diffusion artifacts and the intensity of immunohistological reactions is superior to that obtai...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90365-9
更新日期：1982-01-01 00:00:00
abstract：:A simple, rapid and reproducible method is presented for direct determination of the substitution ratio of a carrier protein with a synthetic nonradioactively labeled peptide. The peptide was covalently linked by a thiol group of a cysteine residue to the immunogenic carrier protein using the heterobifunctional reagen...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90421-8
更新日期：1988-04-06 00:00:00
abstract：:A clonogenic assay system for thymic epithelial cells (TECs) is of crucial importance for identifying thymic epithelial stem and/or progenitor cells, evaluating their activities, and understanding the mechanisms of thymic involution. However, current systems are not sufficiently sensitive at detecting and quantifying ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.02.003
更新日期：2019-04-01 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：:A radioimmunoassay (RIA) for antibodies to the Ro/SSA particle is described using Iodogen to radiolabel the antigenic protein moiety of Ro/SSA with 125I. RIA methods utilizing either 33% saturated ammonium sulfate (NH4)2SO4 or 3.5% polyethylene glycol to separate bound from free antigen are comparable. Either method i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90240-v
更新日期：1990-07-20 00:00:00
abstract：:The development of MHC tetramer staining has opened the doors to multiple avenues of new research [Science 274 (1996) 94]. In this review, we will discuss the development and application of in situ MHC tetramer (IST) staining. We describe two independently developed IST staining methodologies and discuss current uses,...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(02)00197-7
更新日期：2002-10-01 00:00:00
abstract：:The technique of high performance electrophoretic chromatography (HPEC) has been used to fractionate soluble antigens from adult Schistosoma mansoni worms on the basis of molecular weight (MW), prior to screening for their ability to stimulate T lymphocyte activity. Approximately 250 micrograms of protein were separat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90183-8
更新日期：1993-04-02 00:00:00
abstract：:There are several double immunolabelling methods but each has its drawbacks. More often than not, antibodies with the required specificities are available in only one species and their use normally produces false labels due to cross-reactivity. We describe a new and reliable technique for staining with primary antibod...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00281-2
更新日期：1996-04-19 00:00:00
abstract：:A radioimmunoassay for phenobarbital has been studied. Antiphenobarbital antisera were obtained by repeated immunization of rabbits with p-succinamidophenobarbital conjugated to bovine serum albumin. Less than 0.2 pmol of phenobarbital could be measured by this procedure. The specificity of the antibodies was directed...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90327-2
更新日期：1979-01-01 00:00:00
abstract：:A flow cytometric method for enumeration of intracytoplasmic antibody-containing cells was developed. Flow cytometry was found to yield results comparable to traditional fluorescence microscopy in cells stimulated to produce intracytoplasmic antibody by either pokeweed mitogen or Epstein-Barr virus. This technique off...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90083-8
更新日期：1984-02-24 00:00:00
abstract：:Cytolytic T cells (CTL) play a critical role in providing protection against the liver stage of malaria infection. Previous investigations have shown that induction of CTL against peptide or proteins can be achieved by attachment of lipids. In the present study, we used the Plasmodium berghei circumsporozoite protein ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00052-c
更新日期：1995-06-09 00:00:00
abstract：:The short-term kinetics of T cell-mediated cytotoxicity was investigated using a cloning inhibition assay. Murine cytotoxic thymus-derived lymphocytes (CTL) generated in vitro in mixed leukocyte cultures (MLC), were incubated for various periods of time at 37 degrees C with allogeneic mastocytoma target cells. The mix...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90201-0
更新日期：1977-01-01 00:00:00
abstract：:A plasmid cassette vector was designed to generate transgenic mice expressing reporter cDNAs at high levels in antigen-presenting cells under the control of the murine invariant chain (Ii) promoter. Analysis of several transgenic mice harboring a chimeric Ii cDNA placed in this vector showed that it can drive expressi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00276-3
更新日期：2000-11-01 00:00:00
abstract：:Precipitation with polyethylene glycol 6000 is a satisfactory technique for "bound from free" separation in gastrin radioimmunoassay (RIA). This reagent, however, cannot be used in stoichiometric invertase-peptide conjugate-based enzyme immunoassays. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90306-3
更新日期：1980-01-01 00:00:00
abstract：:The chemiluminescence (CL) assay has been used to measure the reactive oxygen species (ROS)-generating capacity of phagocytes. To achieve more optimal measurement conditions for a multi-channel microplate photon-counting CL analyzer with the cooled charge-coupled device (CCD) camera which offers enhanced sensitivity, ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00159-2
更新日期：1997-12-15 00:00:00
abstract：:Paraneoplastic neurological syndromes (PNS) are devastating neurological disorders secondary to cancer, associated with onconeural autoantibodies. Such antibodies are directed against neuronal antigens aberrantly expressed by the tumor. The detection of onconeural antibodies in a patient is extremely important in diag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.02.017
更新日期：2013-05-31 00:00:00
abstract：:We have recently shown that the semi-continuous cultivation of a mouse hybridoma line in spinner flasks, with a basal defined medium (BDM) devoid of serum and protein, increases the secretion of the immunoreactive monoclonal antibody (MAb) by a factor of ca. 2.4, compared to culture in flasks with serum-containing med...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90446-3
更新日期：1990-05-25 00:00:00
abstract：:A sensitive in situ hybridization procedure using both digoxigenin and 35S-labeled riboprobes is described that allows detection of single T cells expressing cytokine mRNA species in both single and double label formats. Modifications to existing procedures have been developed that allow in situ hybridization to be pe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00027-8
更新日期：1995-05-11 00:00:00
abstract：:Pathological prion protein (PrPSc) which is a conformational isoform of a host-encoded protein designated (PrPC) serves as a specific marker protein for the immunochemical diagnosis of transmissible spongiform encephalopathies (TSE). The generation of suitable antibodies to PrPSc therefore underlies the specificity an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00121-x
更新日期：1997-08-22 00:00:00
abstract：:A sensitive assay, using [14C]lecithin as a substrate, has been developed for the measurement of phospholipase activity in rat peritoneal polymorphonuclear leukocytes. Cell extracts were found to contain a phospholipase D activity and indirect evidence suggested that eosinophils are responsible for the cleavage of lec...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90212-4
更新日期：1980-01-01 00:00:00
abstract：:Autoimmune (Hashimoto's) thyroiditis is a chronic inflammatory disease which affects >3% of the population and shows an increasing prevalence with increasing age. Anti-thyroid autoantibodies, particularly against thyroperoxidase (also known as thyroid peroxidase or TPO), occur commonly in humans with autoimmune thyroi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.04.002
更新日期：2007-06-30 00:00:00
abstract：:The influence of human hepatocyte growth factor (HGF) on the transforming growth factor beta (TGF-beta) bioassay CCL-64 was examined. HGF induced proliferation of the CCL-64 cells and potently counteracted TGF-beta-induced growth inhibition. HGF was not inactivated by transient acidification to pH 2, a commonly used p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00228-6
更新日期：1996-01-16 00:00:00
abstract：:There is an emerging trend in the pharmaceutical industry to evaluate a variety of surrogate biomarkers in Phase I/II clinical studies with the intention of determining potential activity of drugs early in clinical development. A number of cytokines expressed in pathological conditions are currently being considered a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00543-9
更新日期：2002-02-01 00:00:00
abstract：:The sensitivity of an enzyme-linked immunosorbent assay (ELISA) for human blood group antigens extracted from blood stains with the surfactant, n-octyl-beta-D-glucopyranoside (OBG), at concentrations below the critical micelle concentration can be increased by the introduction of a single freeze-thaw step. The ELISA s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90245-o
更新日期：1992-05-18 00:00:00
abstract：:Immune analysis of polypeptides separated by SDS-PAGE provides a technique useful in the characterisation of complex mixtures. We describe a simple procedure involving immune analysis of fractionated PAGE gels. Polypeptides of purified herpesvirus simplex, HSV 'excreted antigen' and cell culture 'control' preparations...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90474-5
更新日期：1984-03-16 00:00:00
abstract：:Ia molecule (Ek,b beta:Ek alpha or Ak beta:Ak alpha)-containing planar membranes were constructed with cholesterol and a 9:1 molar ratio of the phospholipids dipalmitoyl phosphatidylcholine and dilinoleoyl phosphatidylcholine. This lipid composition was found to be optimal for the stimulation of T cell hybridomas of d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90432-7
更新日期：1987-04-02 00:00:00
abstract：:Flow cytometry offers a promising alternative to the current methods of amniocentesis or chorionic villus sampling (CVS) for fetal cell sorting for prenatal diagnosis. While flow cytometric methods have been greatly improved to be more sensitive at detecting fetal cells within the maternal circulation, there are still...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2010.09.039
更新日期：2011-01-05 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：:Relatively purified populations of helper T cells free of active PFC precursors can be prepared from primed splenocytes by irradiation, hypotonic shock, and filtration over glass wool. These purified populations can be stored and helper activity recovered undiminished after up to 6 months of cryopreservation. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90082-0
更新日期：1978-01-01 00:00:00
abstract：:A microplate enzyme-linked immunosorbent assay (microELISA) for the study of immunochemical relationships between rabbit myosin light chains is described. Purified individual fast-muscle myosin light chains (LC1F, LC2F and LC3F) and their respective antisera, obtained in chicken, were used. Optimal conditions for anti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90289-8
更新日期：1979-01-01 00:00:00
