Abstract：
:Plasma cells (PCs) secrete antibodies and play an essential role in protective immunity, but also in pathogenesis of antibody-mediated diseases. Physiologically, PCs mainly reside within bone marrow and spleen. In autoimmune diseases such as systemic lupus erythematosus (SLE) autoantibody-producing PCs can also be found at sites of inflammation, e.g. in nephritic kidneys. Therefore, efficient methods are required to reliably analyze and compare PCs at different sites. Flow cytometry and ELISpot analyses are frequently employed for PC characterization and require the preparation of single cell suspensions. To that end, enzymatic digestion is commonly used to isolate immune cells from solid organs like kidneys, occasionally also from lymphoid organs. In this study we show that enzymatic digestion using collagenase may lead to a loss of certain surface markers, e.g. the PC markers CD138 and CD267 (TACI). Therefore, we established an optimized protocol for preparing renal single cells by merely applying mechanical tissue disruption. Omitting enzymatic digestion, this method enables a reliable characterization of viable renal PCs by flow cytometry and cell sorting. We further show that mechanic cell preparation is favorable for lymphocytic immune cell enrichment, while enzymatic disruption improves the yield of digitating or stroma cell populations.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Schaffer S,Maul-Pavicic A,Voll RE,Chevalier Ndoi
10.1016/j.jim.2019.06.019subject
Has Abstractpub_date
2019-11-01 00:00:00pages
112628eissn
0022-1759issn
1872-7905pii
S0022-1759(19)30063-8journal_volume
474pub_type
杂志文章abstract：:A cell ELISA for human leukocytes has been established and standardized. The assay was based on the use of air-dried and methanol-fixed cells which were attached to microplate wells. Cellular antigens were measured through the subsequent binding of monoclonal antibody (MoAb), biotinylated sheep Ig against mouse IgG an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90219-x
更新日期：1986-11-20 00:00:00
abstract：:A method for bleeding rats from the tail vein was developed. Sufficient vasodilation of the tail was accomplished by the use of a specially designed warming device. With the help of a physical restrainer, up to 5 ml of blood can be collected from the tail vein and no anaesthesia is required. This bleeding procedure is...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90148-x
更新日期：1982-09-17 00:00:00
abstract：:Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer antibody therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of antibody drug conjugates (ADCs), efficient removal of surface receptors from cancer cells and changing the pharmacokinetics profile of the...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.02.001
更新日期：2016-04-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.12.007
更新日期：2011-02-28 00:00:00
abstract：:We have developed a sensitive and flexible method for the qualitative evaluation of IgG-Fc receptor interactions in cell suspensions. The assay is based on the flow cytometric quantitation of antibody-coated erythrocyte (EA) rosetting using fluorescein-labelled indicator erythrocytes (E). The number of IgG molecules o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90070-c
更新日期：1990-03-09 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：BACKGROUND:Dendritic cells (DCs) are the most potent antigen-presenting cells. They can be generated in vitro from CD14+ cells, and also from CD34+ progenitor cells. Although T cell proliferation using [3H] thymidine incorporation assay has been used widely to check DC function, this technique only provides limited inf...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00002-4
更新日期：2003-04-01 00:00:00
abstract：:Vaccination against drugs of abuse shows efficacy in animal models, yet few subjects achieve effective serum antibody titers in clinical studies. A barrier to translation is the lack of pre-vaccination screening assays that predict the most effective conjugate vaccines or subjects amenable to vaccination. To address t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2014.01.010
更新日期：2014-03-01 00:00:00
abstract：:We have developed a sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of human heart type fatty acid-binding protein (H-FABP) in human plasma and urine using the combination of two distinct monoclonal antibodies (MAbs) directed against human H-FABP purified from human heart muscle. The total ass...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00248-u
更新日期：1995-01-13 00:00:00
abstract：:Mouse models of allergic asthma are increasingly used to study the immunopathology of this complex disorder. The degree and type of airway inflammation is often studied by determination of differential cell counts on cytospins of bronchoalveolar lavage fluid (BALF) cells stained with May-Grünwald Giemsa, in which the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.03.004
更新日期：2004-05-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90317-z
更新日期：1993-09-15 00:00:00
abstract：:The aim of this study was to establish a cytokine-free, serum-free system which would enable the long-term survival and proliferation of human peripheral blood monocytes. Monocytes were isolated from peripheral blood mononuclear cells (PBMC) by adherence to untreated plastic petri dishes and maintained up to 6 weeks i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90323-l
更新日期：1992-08-30 00:00:00
abstract：:Comparing the marker enzymes horseradish peroxidase (HRP), alkaline phosphatase (AP) and beta-galactosidase (beta Gal) in IgG-coupled form with respect to their temperature-dependent kinetics over a period of 22 h the temperature of 37 degrees C warrants highest substrate turnover for all enzymes at all reaction times...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90388-6
更新日期：1985-05-10 00:00:00
abstract：:A fast and simple indirect immunoperoxidase staining technique is described, which can also be used for the characterization of cells with high endogenous peroxidase activity such as many haemopoietic cells. It is based on the combination of a newly developed glucose-oxidase plus glucose procedure for the inhibition o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90267-x
更新日期：1986-01-22 00:00:00
abstract：:The reactivity of monoclonal antibodies (Mabs) raised against a cyclic peptide representing a chimeric V3 loop of HIV-1 gp120 with different peptide analogues was studied with a biosensor system (BIAcore) and by ELISA. In both assays, the Mabs cross-reacted extensively with the V3 regions of different HIV-1 strains an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90316-6
更新日期：1994-12-02 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90130-3
更新日期：1979-01-01 00:00:00
abstract：:Different Percoll density gradients were used to purify granulocytes from Long-Evans, Fischer, Sprague-Dawley, and Fischer/Long-Evans hybrid rats. Three different discontinuous gradient types were developed which permitted the separation of polymorphonuclear leukocytes (PMNLs) from the peripheral blood of the differen...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90093-0
更新日期：1994-01-03 00:00:00
abstract：:Staphylococcus aureus causes a variety of infections. Knowledge about the physiological role of most S. aureus antigens in colonization and infection is only limited. This can be studied by measuring antigen-specific antibody responses. In this study, we optimized the multiplex microsphere bead-based flow cytometry te...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.12.013
更新日期：2011-02-28 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：:We report a procedure for optimisation of Western blotting using protein G-horseradish peroxidase (protein G-HRP) which avoids the false positive reactions often caused by second antibodies and increases the detection of autoantibodies by protein G conjugate. A number of modifications were investigated. Higher concent...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00235-9
更新日期：1997-03-28 00:00:00
abstract：:Many labs wish to measure cytokines in an accurate, reproducible, and rapid manner. An antibody-based membrane array for measuring cytokines has been developed based on the same technology as the traditional ELISA. The aim of this study was to compare results obtained with the traditional ELISA method with those from ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.10.011
更新日期：2004-01-01 00:00:00
abstract：:Ten solid-phases were evaluated for their usefulness in a two-site immunoradiometric assay for serum thyroid-stimulating hormone. The criteria used to assess each reagent included the minimum detection limit attainable, the change of binding over the concentration range (signal: noise ratio), and ease of preparation o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90491-1
更新日期：1987-11-23 00:00:00
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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：:Recently the analytical power of the latest high throughput next generation DNA sequencing platforms has been used to analyse phage that have been selected from the panning of large combinatorial libraries displaying either peptide or antibody ligands. This process, commonly referred to as next generation phage displa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.03.005
更新日期：2015-05-01 00:00:00
abstract：:With the use of a panel of monoclonal antibodies against structurally related carbohydrate antigens, finely regulated cellular phenotype of the carbohydrate antigens can be characterized. As a model system, the expression of N-acetyllactosamine-related antigens in human biliary epithelial cells was investigated using ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90364-x
更新日期：1988-09-13 00:00:00
abstract：:A new radioimmunoassay has been developed for measuring IgM and IgG rheumatoid factors. Diluted sera from donors and patients were incubated with immunoprecipitates prepared from sheep serum and rabbit anti-sheep IgG antiserum. The precipitates were washed, and radioiodinated rabbit F(ab')2 fragments specific from hum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90259-3
更新日期：1981-01-01 00:00:00
abstract：:Two sets of expression vectors were constructed that permitted the efficient expression of single-chain Fv fragments (scFvs) fused N-terminally to an enhanced mutant of the green fluorescent protein GFP+ or the red fluorescent protein DsRed in insect and mammalian cells. The vectors allowed rapid cloning of scFv fragm...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.12.001
更新日期：2004-02-15 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：:In the current study we attempted to evaluate the suitability of T7 Select 10-3b and lambdaKM8 phage display systems for the identification of antigens eliciting B cell responses in cancer patients and the production of phage-displayed antigen microarrays that could be exploited for the monitoring of autoantibody prof...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.01.022
更新日期：2008-05-20 00:00:00
abstract：:CD4(+) T cells play a central role in regulating the immune response. Their effector function is commonly assessed by their capacity to secrete cytokines detected by ELISPOT and intracellular cytokine staining. However, one aspect of their effector function that is often overlooked is their ability to help activation ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.10.013
更新日期：2013-01-31 00:00:00
