Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1991-11-13
1996-12-10
LeGuyader, John L.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
4352401, 4352402, C12N 508, C12Q 170
Patent
active
055829678
DESCRIPTION:
BRIEF SUMMARY
This invention relates to the human cell line LC5, as well as to the use of same in the investigation of replication inhibition in retroviruses. In particular, this invention is concerned with the investigation of inhibitors and neutralizing antibodies for immunodeficiency viruses of human beings (HIV) and primates (SIV) through use of the human cell line LC5.
The in-vitro investigation of inhibitors for retroviruses has increased in importance in recent years, particularly in the area of AIDS research. It is the goal of this research to develop a therapy for HIV-infected persons. The investigations of neutralizing antibodies are especially interesting with respect to prognosis criteria for investigating the course of HIV infections and for the production of vaccines. In both areas of inquiry--both therapy as well as vaccination--a simple as possible technique for quantifying of the number of infectious viral units is imperative.
Up to now for the in-vitro investigation, cell lines susceptible to retroviruses were utilized, the cells of which grow in a medium. Such a liquid medium with the cell line which is infected with retro-viruses, for example HIV-1-strains, cannot immediately be drawn off, i.e. pipetted off, as the cells float in the medium and consequently could be drawn off along with the liquid medium. Hence, the medium must be allowed to stand some time until the infected cells have settled, so that the liquid medium produced in the meantime can be removed. But, even so, it is always possible that individual cells will again be agitated through the drawing off and are removed together with the liquid medium. Hence, the evaluation of how many cells have become infected by the retrovirus leads to inaccurate test results. Beyond that, a technique of this type is associated with an increased expenditure of work and time.
The large number of sera and substances coming into consideration that may be suitable as inhibitors, makes a rapid and reliable screening procedure necessary. This should be achieved, by way of example, in such a manner, that it could be automated. The use of cell lines in media permits no automation of the procedure, because of the above noted disadvantages.
The object of the invention is to disclose a cell line, that is susceptible to retroviruses and which permits a rapid and reliable investigation of sera and substances with regard to their suitability as inhibitors for retroviruses. In particular, an economical and time saving automation of such an investigation should be possible.
The object of the invention is achieved through the cell line LC5, which is susceptible to retroviruses, in particular to immunodeficiency viruses of human beings (HIV) and primates (SIV) and which can be infected with these viruses. The human cell line LC5 according to the invention is thus particularly suited for the investigation of inhibitors for retroviruses of this type. According to the invention, the use of this new cell line LC5 for the investigation of neutralizing sera and inhibitors for retroviruses is, thus, proposed. The invention further comprises the features of the dependent claims.
The cell line LC5 originates from the lung tissue of a human embryo (E.V. Dantes and V.S. Bolin Fed. Proc. 19, 386, 1960). The original human cell line is deposited under the designation L 132 in the American Type Culture Collection (ATCC, Nr. CCL-5). The cell line L 132 was repeatedly cultured, treated with antibiotics against mycoplasm and subcloned, by which means the cell line LC5 was obtained.
The cell line LC5 according to the invention was deposited on Mar. 9, 1989 in the National Collection of Cultures of Micro-organisms (CNCM), Paris, under Deposit Number I-842.
The human cell line LC5 according to the invention grows by adhering to plastic or glass surfaces, and thus the investigation of inhibitors for retroviruses can be automated in a problem free manner. Owing to the fact that the cell line grows by adhering to plastic and glass surfaces, there is no longer the danger that, with the drawing o
REFERENCES:
American Type Culture Collection Catalogue Of Cell Lines And Hybridomas. Fifth Edition, 1985. p. 5, ATCC CCL5, L-132.
M. J. Tocci et al (1984) Antimicrobial Agents And Chemotherapy vol. 25, No. 2:247-252.
Erfle Volker
Mellert Werner
GSF Gesellschaft fur Strahlen-und Umweltforschung
LeGuyader John L.
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