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1. WO2005100553 - METHOD FOR ASSESSMENT OF CYTOTOXIC LYMPHOCYTE ACTIVITY

Publication Number WO/2005/100553
Publication Date 27.10.2005
International Application No. PCT/US2005/013120
International Filing Date 14.04.2005
IPC
C12N 5/0783 2010.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
07Animal cells or tissues
071Vertebrate cells or tissues, e.g. human cells or tissues
078Cells from blood or from the immune system
0783T cells; NK cells; Progenitors of T or NK cells
G01N 33/50 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
CPC
C12N 2503/00
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
2503Use of cells in diagnostics
C12N 5/0636
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor;
06Animal cells or tissues; Human cells or tissues
0602Vertebrate cells
0634Cells from the blood or the immune system
0636T lymphocytes
G01N 33/5005
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
5005involving human or animal cells
G01N 33/505
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
5005involving human or animal cells
5008for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
5044involving specific cell types
5047Cells of the immune system
505involving T-cells
Applicants
  • BOARD OF SUPERVISORS OF LOUISIANA STATE UNIVERSITY AND AGRICULTURAL AND MECHANICAL COLLEGE [US]/[US] (AllExceptUS)
  • CUI, Yan [CA]/[US] (UsOnly)
  • CHEN, Kong [CN]/[US] (UsOnly)
Inventors
  • CUI, Yan
  • CHEN, Kong
Agents
  • DAVIS, Bonnie, J.
Priority Data
60/563,04316.04.2004US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) METHOD FOR ASSESSMENT OF CYTOTOXIC LYMPHOCYTE ACTIVITY
(FR) PROCÉDÉ D'ÉVALUATION DE L'ACTIVITÉ LYMPHOCYTAIRE CYTOTOXIQUE
Abstract
(EN)
A new cytotoxic T lymphocyte (CTL) assay has been discovered using two cell lines that stably express either green fluorescent protein (GFP) or red fluorescent protein (DsRed), which are distinguishable by FACS, fluorescence microplate reader, or fluorescence microscopy. Using one cell line as a target (T) to present antigen and the other, at the same number, as an internal control (reference, R), a new CTL assay (named fluorolysometric (FL)-CTL assay) way developed based on cytolysis of these fluorescent protein-expressing targets detectable by FACS. This FL-CTL assay was further extended for use with a fluorescent microplate reader. This FL-CTL assay was reproducibly used to determine primary CTL activity at high sensitivity when compared to other conventional assays with in vivo activated T cells against different antigens. This new reliable, sensitive, convenient, and economical CTL assay has broad application potentials for experimental and clinical use in different antigen and effector-target systems.
(FR)
La présente invention a trait à un dosage lymphocytaire T cytotoxique (CTL) au moyen de deux lignées cellulaires d'expression stable de protéine fluorescente verte (GFP) ou de protéine fluorescente rouge (DsRed) qui sont aptes à être distinguées par un trieur de cellules marqué par fluorescence (FACS), un lecteur pour microplaques par fluorescence, ou la microscopie par fluorescence. En utilisant une lignée cellulaire comme cible (T) pour la présentation de l'antigène et l'autre, au même numéro, comme contrôle interne (R), on a développé un nouveau type de dosage (désigné dosage lymphocytaire T cytotoxique lysométrique par fluorescence, FL-CTL) basée sur la cytolyse de ces cibles d'expression de protéines fluorescentes détectables par trieur de cellules marqué par fluorescence. Ce dosage lymphocytaire T cytotoxique lysométrique par fluorescence a été développé davantage pour être utilisé avec un lecteur pour microplaques par fluorescence. Ce dosage lymphocytaire T cytotoxique lysométrique par fluorescence a été utilisé de manière reproductible pour la détermination de l'activité lymphocytaire T cytotoxique à sensibilité élevée par rapport aux autres dosages classiques avec des lymphocytes T activés in vivo contre différents antigènes. Ce nouveau dosage lymphocytaire T cytotoxique fiable, sensible, simple et économique présente de vastes potentiels d'applications pour une utilisation expérimentale et clinique dans divers systèmes d'antigènes et de cibles effecteurs.
Also published as
US2007224660
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