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Calcein-acetyoxymethyl cytotoxicity assay: standardization of a method allowing additional analyses on recovered effector cells and supernatants.
Clin Diagn Lab Immunol
November 2001
Dipartimento di Medicina Interna e Gastroenterologia, University of Bologna, Bologna, Italy.
Cytotoxicity assays provide an in vitro evaluation of the lytic activity of NK and T cells against tumors or transformed cells. However, none of these methods allow the recovery of cells or supernatants after the assay. We standardized a microcytotoxicity test using calcein-acetoxymethyl (calcein-AM) dye that requires very small quantities of cells while maintaining the same sensitivity as the traditional (51)Cr assay.
View Article and Find Full Text PDFMeasurement of NK activity by the microcytotoxicity assay (MCA): a new application for an old assay.
J Immunol Methods
July 2001
Department of Pharmacology, University of Pittsburgh School of Medicine, 15213, Pittsburgh, PA, USA.
Natural killer (NK) cells are spontaneously cytotoxic immune effector cells with the ability to selectively destroy tumor cells without harming normal cells. To perform this function, NK cells utilize two main cytotoxicity pathways, the well known perforin/granzyme-mediated secretory/necrotic killing and the recently defined TNF family ligand-mediated non-secretory/apoptotic killing. The former mechanism is manifested mainly against a few cultured leukemia cell targets, while the latter mediates killing against a large variety of tumor cell targets.
View Article and Find Full Text PDFA reliable, rapid and inexpensive two-color fluorescence assay to monitor serum cytotoxicity in xenotransplantation.
J Immunol Methods
January 1999
Oklahoma Transplantation Institute, Integris Baptist Medical Center, Oklahoma City, USA.
Removal and/or neutralization of preformed anti-pig antibodies in non-human primate blood have been shown to prevent the hyperacute rejection of transplanted pig organs. The purpose of this study was to establish a suitable in vitro method that would allow for screening and comparison of various agents and methods potentially useful in the prevention of hyperacute rejection. The pig kidney cell line (PK15), pig aortic endothelial cell line (AG08472), and a primary culture of endothelial cells explanted from a pig aorta were incubated with either human or baboon sera.
View Article and Find Full Text PDFUse of [35S]methionine-labelled rat lymphoblasts in micro-cytotoxic and limiting dilution assays.
J Immunol Methods
June 1996
Axe Immunologie, Laboratoires Fournier, Daix, France.
When only limited numbers of effector cells are available for in vitro T cytotoxic determinations, standard assays cannot be performed. 51Cr is still the most commonly used marker of target cells in cytotoxicity assays but since the incorporation of this marker is low, especially in non-tumor cells such as lymphoblasts, larger numbers of both target and effector cells are required. Here we report the use of [35S]methionine-labelled rat ConA blasts in cytotoxic, micro-cytotoxic and limiting dilution assays.
View Article and Find Full Text PDFStandardization of a micro-cytotoxicity assay for human natural killer cell lytic activity.
J Immunol Methods
June 1994
Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla Putti I.O.R., Bologna, Italy.
Cytotoxicity assays are widely used to evaluate the functional activity of NK and T cells against tumour target cells and the release of radioactive sodium chromate from labelled target cells is still the most commonly used marker of target lysis in culture supernatants. We describe here the standardization of a micro-cytotoxicity test in which the number of cytolytic effector and tumour target cells have been decreased by a factor of 10. The release obtained by 500 tumour target cells was compared with the release obtained by 5000 target cells in the standard cytotoxicity assay for target:effector cell ratios from 1:1 to 1:100.
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