Abstract：
:Natural killer (NK) cells are important mediators of virus- and tumor-specific immune responses. The transfection of genes into NK cells has been proven difficult and so far requires infection with virus-based vectors. Here, the application of a novel nonviral, electroporation-based gene transfer method is described for the rapid and highly efficient transient transfection of NK cell lines as well as freshly isolated NK cells. In contrast to conventional methods, this technique, termed nucleofection, leads to direct transfer of DNA into the nucleus. Using reporter proteins H-2K(k), luciferase+, and enhanced yellow green fluorescent protein (EYFP) as independent read-out systems, transfection efficiencies of well over 50% were achieved in transient transfection assays. The highest luciferase activity could be measured only 4 h after transfection, whereas EYFP, when analyzed by flow cytometry, showed expression peaks after 28 h. Interestingly, best transfection efficiencies were achieved with non-dividing NK cells. The novel nuclear gene transfer method presented here is highly useful for the analysis of NK cell-specific gene regulation and should facilitate the development of NK cell-based gene therapy approaches.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Trompeter HI,Weinhold S,Thiel C,Wernet P,Uhrberg Mdoi
10.1016/s0022-1759(02)00431-3keywords:
subject
Has Abstractpub_date
2003-03-01 00:00:00pages
245-56issue
1-2eissn
0022-1759issn
1872-7905pii
S0022175902004313journal_volume
274pub_type
杂志文章abstract：:The Raji cell assay for circulating immune complexes (CIC) is frequently the method of choice when the detection of large, complement fixing complexes is desired. We have developed an enzyme-linked immunosorbent assay modification (Raji-ELISA) of the Raji cell technique which is easy to perform, uses commercially avai...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90046-7
更新日期：1983-01-01 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：:Antigen-specific lymphocytes are important in the immune response to viral infection. Peripheral blood mononuclear cells (PBMC) are traditionally used as a source of effector cells in most immunological studies. We described here the use of the bispecific monoclonal antibodies (BSMAB) anti CD3:CD8 (CD3,8) and anti CD3...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00510-0
更新日期：2003-03-01 00:00:00
abstract：:Helper and suppressor activity of T cell lines were investigated using an assay system which is based on measurement by the enzyme-linked immunosorbent assay (ELISA) of the antibody produced in vitro. T cell lines were established from spleens of BALB/c mice immunized with ovalbumin and human serum albumin. The T cell...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90063-7
更新日期：1983-02-25 00:00:00
abstract：:High-titered antisera against human melanoma- and leukemia-associated antigens have been produced in non-human primates. The data from this and from other studies suggest that this model may prove a valuable source of highly specific antisera against a variety of human tumor-associated antigens or alloantigens. Our me...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90048-x
更新日期：1976-01-01 00:00:00
abstract：:The IgG binding domains of staphylococcal protein A and streptococcal protein G were expressed as a chimaera using the pGEX vector which has been advocated because its fusion proteins tend to be soluble and easily isolated on immobilised glutathione. This chimaera was soluble and abundant (yield = 18 mg/l of bacterial...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90087-a
更新日期：1992-07-31 00:00:00
abstract：:Differentiating foot-and-mouth disease virus (FMDV) antibodies generated during a natural infection from those due to vaccination (DIVA) is crucial for proving freedom from disease after an outbreak and allowing resumption of trade in livestock products. The World Organisation for Animal Health (OIE) recommends that F...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.11.004
更新日期：2010-01-31 00:00:00
abstract：OBJECTIVE:The central mechanism for establishing a self-tolerant and functional T cell repertoire includes the promiscuous expression of otherwise tissue-restricted proteins by medullary thymic epithelial cells (TEC). We here demonstrate a novel and highly efficient method for isolating this rare key cell type. METHOD...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.02.001
更新日期：2019-04-01 00:00:00
abstract：:A method is presented for measuring the hemolytic activity of undiluted guinea pig serum. Undiluted serum is placed in contact with a column of sensitized sheep red blood cells in an agar matrix in a capillary tube. After incubation, the length of the zone of lysis is measured. Comparison with a standard curve prepare...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90212-6
更新日期：1979-01-01 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：:Monoclonal and anti-dinitrophenyl and anti-trinitrophenyl IgE antibodies were used to measure heterocliticity using competitive inhibition assays with homologous and heterologous haptens. The antibodies or antibody-containing ascites fluids were diluted to give 50% of the maximum binding to wells of antigen-coated mic...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90516-3
更新日期：1987-11-23 00:00:00
abstract：:A new adhesion assay was developed that utilizes buoyancy, rather than washing or centrifugation, to remove non-adherent cells. Biotinylated cells were added to wells containing cell monolayers or purified protein substrates. Non-adherent cells were then removed by floatation on a dense Percoll solution. The adherent ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00187-6
更新日期：1995-12-01 00:00:00
abstract：:Purified polyclonal immunoglobulin preparations representing the 4 rat IgG subclasses were tested for binding to Staphylococcus aureus protein A attached to 3 different solid phases (Staphylococcus aureus Cowan I bacteria, Sepharose CL4B and Sepharose 6MB). Protein A Sepharose CL4B showed higher reactivity with IgG1 a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90251-x
更新日期：1983-08-26 00:00:00
abstract：:Incorporation of control valves into a previously described device enabled us to regulate the formation of 8 suction blisters on the upper surface of the forearm in adult human volunteers. After the removal of the raised epidermis and blister fluid, uniform areas of denuded dermis were obtained by placing hollow adhes...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90248-0
更新日期：1985-11-07 00:00:00
abstract：:The paper by Herzyk et al. which appears in this issue of the Journal of Immunological Methods is an important and long overdue contribution. The study clearly and convincingly demonstrates that two widely used commercial ELISA kits fail to detect over 90% of the 31 kDa human IL-1 beta precursor (pro-IL-1 beta) in var...
journal_title：Journal of immunological methods
pub_type： 评论,杂志文章
doi：10.1016/0022-1759(92)90179-w
更新日期：1992-04-08 00:00:00
abstract：:The attachment of cells to plastic tissue culture plates is a widely used criterion of cell viability in microcytotoxicity assays. When cell suspensions of primary human colon carcinoma and melanoma cells which were of low initial viability (assessed by trypan blue exclusion) were allowed to adhere to tissue culture p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90039-9
更新日期：1981-01-01 00:00:00
abstract：:There has been recent interest in the depletion of regulatory T cells (Tregs) as part of a multi-faceted approach to the immunotherapy of melanoma patients. This is in part due recent findings that convincingly show that Tregs are an integral part of regulating and even suppressing an immune response to growing tumor ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.01.012
更新日期：2008-04-20 00:00:00
abstract：:Tetramolecular monoclonal antibody complexes were used to selectively cross-link a subset of human peripheral blood T cells (CD8 positive) to glass beads coated with hapten (fluorescein) modified bovine serum albumin. Tetramolecular antibody complexes were prepared with anti-CD8 (Leu 2a), anti-FITC mouse IgG1 monoclon...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90361-4
更新日期：1988-09-13 00:00:00
abstract：:IgA antibodies are key immune effectors against invading pathogens but also possess essential immunoregulatory functions. Detecting and quantifying human IgA+ B-cell subsets and secreted IgA molecules is needed for investigating the protective, modulatory and pathophysiologic roles of IgAs. Here, we produced a recombi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112833
更新日期：2020-01-01 00:00:00
abstract：:Enzyme-linked antibody was used to treat unfixed herpesvirus-infected human fetal lung cell cultures in a mode which permitted the visualizing of local sites of infection. Foci containing as few as 20 herpesvirus-infected cells produced sufficient viral mass to be easily detectable by this method. 'Clouds' or 'plumes'...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90361-6
更新日期：1981-01-01 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：:An antibody binding-inhibition test is described, which allows the detection of P. falciparum in red blood cells (RBC) infected in vitro, using a crossreacting, monoclonal anti-P. berghei antibody and P. berghei coated microtiter plates. Experiments carried out to determine the coating efficiency of various P. berghei...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90231-5
更新日期：1985-09-03 00:00:00
abstract：:The histamine content of commercial preparations of pokeweed mitogen was measured by amino acid analysis technique, automated fluorometry, and bioassay employing the guinea pig ileum. Ten samples from 5 companies were examined and found to contain between 0.026 micrograms and 167.5 micrograms of histamine per ml of so...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90095-3
更新日期：1982-12-30 00:00:00
abstract：:A specific and sensitive enzyme immunoassay system for human secretory IgA was developed using anti-alpha-chain antibodies coupled to activated thiol-Sepharose, and anti-secretory component antibodies labeled with beta-D-galactosidase from Escherichia coli. The dose response of the enzyme activity in eluate was observ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90053-4
更新日期：1983-01-01 00:00:00
abstract：:C3 nephritic factor (C3NeF) is an autoantibody against the C3 convertase which stabilizes this otherwise inherently labile neoenzyme and induces a continuous activation of the alternative pathway with C3 depletion. NeF is found in patients with membranoproliferative glomerulonephritis and/or partial lipodystrpohy. NeF...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00295-2
更新日期：2001-05-01 00:00:00
abstract：:Dispase-treated murine hybridoma cells (SP2/0-Ag14) were transfected with the G418 resistance gene bearing plasmid pSV2-neo by electropermeabilization with a high degree of efficiency. The cells were subjected to intermittent multiple high-voltage short duration (5 microsecond) DC pulses at intervals of 1 min in a wea...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90452-8
更新日期：1988-04-22 00:00:00
abstract：:Certain serum samples produce high background in Western and direct immunoblot assays that detect human serum IgG against specific antigens. We determined that this was due to a reaction between endogenous IgG and the membrane blocking agent (we refer to this as blocking-specific background). Using milk as blocking ag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90259-a
更新日期：1993-01-14 00:00:00
abstract：:Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants, which can cause cancer in humans. The maximum tolerable limit of benzo[a]pyrene (B[a]P) in drinking water was set to 10 ng/L by the European Commission (Council Directive 98/83/EC), because of its highly carcinogenic and mutagenic effect ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.016
更新日期：2011-08-31 00:00:00
abstract：:This investigation describes a new procedure for the preparation of immunogenic liposomes which circumvents the need to synthesize the N-(hapten)-substituted derivatives of phosphatidylethanolamine that were previously employed for this purpose. The method is based on the generation of liposomes containing the N-hydro...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90125-4
更新日期：1983-12-30 00:00:00
abstract：:A dot-ELISA technique for the detection of G. lamblia specific antigen in stool eluates of clinical cases of giardiasis was developed and evaluated employing monospecific antibodies to a G. lamblia specific coproantigen with a molecular mass of 66 kDa. The assay detected 22 (91.7%) of the 24 microscopically confirmed ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90030-j
更新日期：1991-03-21 00:00:00
