Abstract：
:Many normal and pathological processes have been shown to be influenced by the quantitative and qualitative nature of cytokine production. Cytokines have also been shown to be central in modulating host responses to invasion by pathogens. Therefore detection and analysis of cytokine production are critical in the understanding of host responses to a variety of immunological insults. The detection of cytokines by bioassay or immunoassay provides data from in vitro experiments and can be used to determine the circulating plasma levels of particular cytokines. However, many investigations require the detection of cytokines in small numbers of cells or directly from tissue samples. It is in this particular area that detection and analysis of cytokine gene expression by use of the sensitive and specific reverse transcription-polymerase chain reaction (RT-PCR) has application. The purpose of these protocols is to provide a practical guide to the detection of cytokine gene expression using RT-PCR techniques. Procedures for analysing mRNA expression in cell culture, cell suspensions and tissue samples will be described and pitfalls and caveats concerning the techniques will be discussed. These protocols should provide a starting point for the development of RT-PCR technology in laboratories with only limited molecular experience.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Walker KBdoi
10.1016/s0022-1759(97)00124-5subject
Has Abstractpub_date
1998-03-01 00:00:00pages
113-23issue
1eissn
0022-1759issn
1872-7905pii
S0022-1759(97)00124-5journal_volume
212pub_type
杂志文章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：:Enzyme-linked immunoadsorbent assay (EIA) has widespread use for the measurement of antibody concentration. The affinity constant (Kaff) of the antibody has an effect upon the quantification by EIA. It is thus important to be able to measure Kaff by solid-phase EIA. Based upon the Law of Mass Action and using serial d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90187-6
更新日期：1987-06-26 00:00:00
abstract：:The proinflammatory cytokine IL-18 mediates IFN-gamma production as well as the induction of Th1 polarized immune responses in synergy with IL-12. In this study, we describe the production of isogeneic monoclonal antibodies (Mabs) directed against murine IL-18 (mIL-18). Immunization of IL-18-deficient mice with recomb...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00505-1
更新日期：2002-01-01 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：: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：INTRODUCTION:Basophils play a major physio-pathological role in hypersensitivity related diseases. Basophils express high affinity Immunoglobulin (Ig) E receptors (FcεRI), IgG and complement regulatory. Basophils also have immunoregulatory activity through interaction with T cells. The aim of this study was to look for...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112951
更新日期：2021-01-22 00:00:00
abstract：:To date, studies on T cells in acute myeloid leukemia (AML) have been limited to flow cytometric analysis of whole peripheral blood mononuclear cell (PBMC) specimens or functional work looking at the impact of AML myeloblasts on normal or remission T cells. This lack of information on T cells at the time of presentati...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.06.010
更新日期：2009-08-31 00:00:00
abstract：:To maximize safety in the setting of an increasing number of requests for flow cytometric analysis of specimens potentially contaminated with the AIDS retrovirus, we evaluated some commonly used fixatives for their ability to inactivate the infectious potential of the virus. We found that both formaldehyde (0.37% v/v)...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90278-4
更新日期：1986-01-22 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) for the quantitation of rat erythrocyte membrane sialoglycoproteins (glycophorins) has been developed. Samples of erythrocytes and reticulocytes were analysed using this assay, and response compared among them and purified glycophorins samples. A broadly homologous behaviou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90387-m
更新日期：1993-06-18 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：:An enzyme immunoassay for the detection of IgG and IgM anti-polymerized albumin autoantibodies (AAA) is described. It was found that polyalbumin receptors on HBsAg particles interfere in the detection of IgG AAA when polymerized human albumin (pHSA), but not polymerized bovine albumin (pBSA), is used as coating antige...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90484-3
更新日期：1985-01-21 00:00:00
abstract：:The possibility that immortalisation of in vitro immunised splenocytes may result in hybridomas secreting polyreactive antibodies was investigated. A panel of nine murine hybridomas, secreting IgM(kappa) anti-goat immunoglobulin G (anti-GIgG), was produced by immortalising splenocytes that had been immunised in vitro ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00481-1
更新日期：2001-12-01 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) using murine spermatozoa and thymocytes is described for the detection of anti-H-Y antibodies. ELISA is more sensitive and reproducible than cytotoxicity and the PA-SRBC test. It is particularly useful for the rapid screening of a large number of samples. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90182-4
更新日期：1981-01-01 00:00:00
abstract：:A polyethylene glycol (PEG) radioimmunoprecipitation assay for human IgG is described that is sufficiently sensitive to detect 0.5 ng of IgG. This model antibody-antigen system was also used to study the stoichiometries of PEG-precipitated complexes. Our results suggest that the presence of PEG may affect the stoichio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90091-7
更新日期：1979-01-01 00:00:00
abstract：:Methods are described for the in vitro generation and detection of antibody-secreting cells (PFC) from channel catfish. Hapten-specific PFC can readily be enumerated by an indirect plaque assay employing rabbit antiserum to catfish Ig and guinea pig complement. A modified Mishell-Dutton-type culture system was develop...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90006-1
更新日期：1984-09-04 00:00:00
abstract：:It was found that 10% horse serum (HrS) could be used to support an in vitro antibody response to SRBC. It could replace the fetal calf serum (FCS) in both the culture medium and the nutritional feed. The response could be improved by the addition of 2-mercaptoethanol or by including both FCS (5%) and HrS (5%) in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90042-3
更新日期：1980-01-01 00:00:00
abstract：:Antibodies to a wide range of self-antigens, including those directed against nucleic acids or nucleic acid-binding proteins are the essential biomarkers for diseases such as systemic lupus erythematosus (SLE). Highly complex libraries of nonamers consisting of N-substituted glycines (peptoids) were screened for compo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.11.004
更新日期：2014-01-15 00:00:00
abstract：:A method for quantitative RT-PCR using ELISA detection was developed and applied to the quantitation of IL-1 beta mRNA in clinical samples. To compensate for the 'tube effect' of RT-PCR, a synthetic RNA, pRSET RNA, was added to sample solutions as an internal standard and co-amplified with IL-1 beta mRNA. Sense primer...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00321-m
更新日期：1995-03-27 00:00:00
abstract：:A competitive RT-PCR method that permits reliable quantification of minute amounts of reverse-transcribed mouse lymph node mRNA is described. Using this technique, an absolute number of cDNA copies ranging from 10(3) to 10(5) can be determined, with a precision superior to 25%. The standard templates described in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00186-5
更新日期：1997-12-29 00:00:00
abstract：:Most monoclonal antibodies to mouse antigens have been derived from rat spleen-mouse myeloma fusions. Many resultant hybridomas express one of several myeloma kappa chain transcripts, even though the parent myeloma may have been ascribed as not expressing light chain protein. Previous reports have only differentiated ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.07.002
更新日期：2006-08-31 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：: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：:A new sandwich-type solid-phase enzyme immunoassay (EIA) for carcinoembryonic antigen (CEA) was established by using two selected monoclonal antibodies (MAbs). One MAb, F82-61, which is immobilized on polystyrene beads, reacts with an epitope present on the N-terminal domain (N) of the CEA molecule and the other, F11-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90388-n
更新日期：1993-06-18 00:00:00
abstract：:Regulatory T cells (Treg) have recently come to the fore in studies of immune regulation, particularly in autoimmune disease and cancer. While there appear to be several distinct subsets of T cells with regulatory function, a population described as natural Treg and characterized by expression of the transcription fac...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.06.006
更新日期：2007-10-31 00:00:00
abstract：:In the field of sexually transmitted infections (STI), the cervicovaginal explant (CVEx) model, not only provides the opportunity to study the different immunological arms present in these tissues under steady state conditions, but also their response against ex vivo infection with relevant pathogens. The methodology ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2018.06.005
更新日期：2018-09-01 00:00:00
abstract：:A new method is described for demonstrating the individual subclasses of human IgG immunoglobulins by means of analytical isotachophoresis. Individual subclasses of IgG purified from sera of multiple myeloma patients have distinct mobility patterns. When a mixture with purified IgG for each of the 4 subclasses is anal...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90163-7
更新日期：1979-01-01 00:00:00
abstract：:Interleukin 2 (IL-2) is a class of T cell growth factors produced by T cells of a number of species. The unique growth-promoting properties of these molecules allow the development of antigen-specific effector T cell lines which can be used to analyze the molecular basis of lymphocyte interactions. A murine T cell tum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90055-1
更新日期：1980-01-01 00:00:00
abstract：:A method to purify cobra venom factor (CVF) from cobra venom by sequential column chromatography is described which yields a product virtually free of phospholipase A2, a common contaminant of CVF preparations. The separation of phospholipase A2 from CVF was achieved by chromatography on Cibacron blue-agarose, a resin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90045-0
更新日期：1984-10-12 00:00:00
abstract：:Anti-drug neutralizing antibodies (NAbs) formed due to unwanted immunogenicity of a therapeutic protein point towards a mature immune response. NAb detection is important in interpreting the therapeutic's efficacy and safety in vivo. In vitro cell-based NAb assays provide a physiological system for NAb detection, howe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.02.006
更新日期：2015-04-01 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
