[Dye exclusion test and LDH release assay (author's transl)].
Trypanblue exclusion test and LDH release assay were compared concerning correlation and sensitivity. Short time cultures (4 h; 37 degrees C) of human blood leucocytes were carried out in the presence of various doses of either different tissue extracts or digitonin and assayed by the described methods. Three different reaction types have been found: 1. synchronic type: In dependence on the used antigen the LDH release and the number of coloured cells both show either an analogous dose-dependent positive tendency or a negative reaction. 2. asynchronic type: In this case the trypanblue exclusion test shows negative results. In contrast an increase of LDH release is determined suggesting a permeability-increasing influence of the used substance. 3. stabilization type: In comparison with the control the release of LDH is clearly suppressed up to a definite concentration of the substance. The trypanblue exclusion test is not able to detect this cell stabilizing effect. The LDH release assay is sensitive, objective and in the case of an automatic measurement very effective. A necessary presumption for the test is the absence of LDH or enzyme activity-influencing factors in the substance itself. Reasons for the three types of reaction were discussed. It is recommended to apply this enzyme release assay to methods basing on cellular in-vitro short-time-cultures of nucleic cells (testing of pharmaca, potential antigens or allergen preparations and bio-materials).
D002470:Cell Survival; D006801:Humans; D007111:Immunity, Cellular; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D007962:Leukocytes / Q000201:enzymology / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods; D014343:Trypan Blue
D007770:L-Lactate Dehydrogenase; D014343:Trypan Blue
Farbstoffexklusions- und LDH-Freisetzungstest.