FIELD: microbiological and immunological methods.
SUBSTANCE: invention is intended for use to study structural and functional arrangement of nucleolus and mechanisms of action of pharmacological preparations on human cells. Mouse hybridoma cell strain, called A3, is obtained through fusing mouse splenocytes immunized by coarse fraction of nuclei of human cell line RAMOS with cells of mouse myeloma line P30X63-Ag8.653. Resulting hybridoma secretes monoclonal antibodies against antigen, called A3 antigen, localized inhuman cell nucleoli irrespective of their tissue or line origin. In cases of different cell fixation ways. A3 antigen is revealed as incorporated in discrete (typically several tens) foci located exclusively in the zone of nucleoli. Unique property of A3 antigen is its high sensitivity to the action of various protein synthesis inhibitors, e.g. emetine, anisomicyne, cycloheximide, and puromicyne. During incubation of cells in presence of above-listed substances, A3 antigen migrates from nucleoli to numerous foci located in nuclear nucleoplasma. This A3 antigen migration precedes apoptotic death of cells. Monoclonal antibodies A3 produced by the strain A3 are recommended to reveal nucleoli and to estimate total level of protein synthesis in human cells by cell biology techniques. Antibodies can be used to reveal possible contamination of human cell cultures with another-species cells as well as to investigate biological activity of known and novel protein synthesis inhibitors, including pharmacological preparations in human in vitro cells.
EFFECT: expanded microbiological and immunological possibilities.
2 cl, 6 dwg
