Process for in vitro analysis of toxic and allergenic substances

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 71, 435 72, 435 721, 435325, 435326, 435372, 4353723, 435373, 435375, 530351, G01N 3353, G01N 33567

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active

060460105

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BRIEF SUMMARY
The present invention concerns a process for in vitro analysis of toxic and allergenic substances. Substances intended for use as pharmaceuticals, food additives, cosmetic or hygienic products, industrial chemicals and other sub- stances are analysed for adverse reactions. Such reactions may be allergic reactions, skin irritating effects and toxic effects.
Today such tests are performed in vivo on animals. Animals are expensive to raise and keep. Further, some tests may involve suffering of the animals.
The present invention provides a process for in vitro analysis of the adverse effects of substances. The analysis of the substances is performed on blood cells from warm blooded animals. No animals are involved since the substances are tested on human cells i.e. cells from the same species as they are intended for. Some substances may not have the same effect on different species, and tests performed on animals may give false results. Therefore the test is preferably performed with human blood.
Thus, the method will be offered as an alternative to animal tests for food additives, cosmetic or hygienic products, industrial chemical, drugs and other substances where adverse reactions are to be avoided.
In vitro evaluation of toxic effects on the immune system has been tested on splenocytes from mice ("In vitro evaluation of drug-induced toxic effects on the immune system as assessed by proliferative assays and cytokine production", M. Pallardy et. al. Eur. Cytokine Net., Vol. 2 No. 3, May-June 1991, pp. 201-206). The method was poorly effective detecting molecules inducing autoimmunity and hypersensitivity.
It has now turned out that according to the present invention it is possible to analyse adverse tissue reactions to foreign substances in vitro by cultivating them with human whole blood, prevented from coagulating and analysing the cytokine produced by the human cells.
Before a substance has been in use and before the target group has been exposed the present method offers the possibility to predict whether a risk for adverse reactions of inflammatory or allergic type will occur.
The invention provides an interpretation key for evaluating the adverse tissue reactions to foreign substances.


SUMMARY OF THE INVENTION

The method is defined in claim 1 and is based on cultivation of blood cells in vitro. The substance to be evaluated is added to the blood culture. If the reactive cells of the blood recognises the test material as foreign or damaging a production of inflammatory signal substances, cytokines occurs. The blood contains different white blood cells with different functions in the response to foreign substances. The different types of inflammatory cells secrete different repertoirs of cytokines. The patterns, (profiles) of the secreted cytokines indicate what compartment of the immune system that has been activated. Thereby is it also possible to predict what kind of adverse reaction to be expected from a substance if administered to humans or experimental animals. Some substances will induce a typical cytokine profile driving the immune system towards IgE mediated allergy, thus indicating a potential of those substances to evoke asthma, hay fever and urticaria. Other substances will induce a cytokine pattern driving the immune system to cellular immunity thus having the potential to result in contact excema as an adverse reaction. Finally, a chemical or toxic effect will result in a cytokine pattern typical of alarm reactions seen in tissue damage and operations. Thus, the concept that the present method is based on is to use an interpretation of the cytokine pattern that a substance can induce in blood in vitro cells to predict whether the substance has the potential to cause allergy of different types or is generally hazardous to induce tissue damaging or toxic effects.
We classify adverse tissue reactions to foreign substances as follows. hypersensitivity and is mediated by IgE antibodies produced specifically to the foreign substance. An acute inflammatory reaction is produced, histamine is of

REFERENCES:
patent: 5439799 (1995-08-01), Rautenberg et al.
McHugh et al, Allergy, vol. 49, pp. 751-759, Oct. 1994.
M. Imada et al., "Allergen-stimulated Interleukin-4 and Interferon-gamma Production in Primary Culture: Responses fo Subjects with Allergic Rhinitis and Normal Controls", Dialog Information Services, File 155, Medline Dialog accession No. 09484227, Immunology, England, Jul. 1995, pp. 373-380.
K. Mueller-Decker et al., "Development of an In-Vitro Alternative Assay to the Draize Skin Irritancy Test Using Human Keratinocyte-Derived Proinflammatory Key Mediators and Cell Viability as Test Parameters", In Vitro Toxicol 5 (4), Dialog Information Services, file 5, Biosis Dialog accession No. 10103327, Biosis accession No. 95103327, 1992, pp. 191-209.
R. V. House et al., "In Vitro Evaluation Fentanyl and Meperidine for Immunomodulatory Activity", Dialog Information Services, file 155, Medline accession No. 09556564, Immunol Letters, The Netherlands, May 1995, pp. 117-124.
C.G. Larsen et al, "The Delayed-type Hypersensitivity Reaction is Dependent on IL-8. Inhibition of a Tuberculin Skin Reaction by an Anti-IL-8 Monoclonal Antibody", Dialog Information Services, file 155, Medline accession No. 09433182, J Immunol, United States, Aug. 15, 1995, pp. 2151-2157.
J.D. Dewitte et al., "Improved HPLC Determination of Urinary Neopterin", Dialog Information Services, file 155, Medline accession No. 06865075, Biomed Chromatogr, England, 1987, 2 (5), pp. 183-188.

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