Transfected Chinese hamster ovary cells as a model system for cytokine immunocytochemistry and in situ hybridisation.
The presence of cytokine producing cells is most easily revealed by techniques measuring the secreted cytokines in culture supernatants or body fluids. However, these techniques only measure the bulk cytokine release by a given, often mixed cell population. To demonstrate cytokine production at the single cell level, immunocytochemistry (ICC) and in situ hybridisation (ISH) are now widely used techniques. To establish these techniques, an easily accessible model system is needed which permits the evaluation of different ICC and ISH protocols. It can be used to demonstrate the specificity of the antibodies and may serve as a positive control for samples of unknown cytokine content. Here we propose the use of Chinese hamster ovary (CHO) cells transfected to express one specific cytokine as such a model system. Its usefulness is demonstrated by the characterisation of six monoclonal antibodies to human interleukin-4 and the establishment of two in situ hybridisation protocols.
Mueller
R
R
Ciba Geigy AG, Basel, Switzerland.
Junker
U
U
Wagner
K
K
Heusser
C
C
Bullock
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D016707:Tissue Fixation / Q000379:methods; D014162:Transfection
D000911:Antibodies, Monoclonal; D016207:Cytokines; D011993:Recombinant Fusion Proteins