Detection of interleukin 1 with human dermal fibroblasts.
A fibroblast proliferation assay was developed for the detection of interleukin 1 (IL 1). Proliferation was measured by thymidine incorporation and by staining of cellular proteins with crystal violet. Response of fibroblasts was optimal at cell numbers of 4,000 to 9,000 cells/culture and an incubation period of four days. Serum content of the culture medium, ranging from 1 to 10% fetal calf serum (FCS), enhanced the proliferative response in a concentration-dependent manner, while higher concentrations of FCS did not lead to further increase. Both detection methods were equally suitable for the measurement of IL 1 biological activity in purified and crude preparations. In contrast to the conventional thymocyte comitogenic assay, the fibroblasts in this assay did not proliferate in response to IL 2 or IL 6. Fibroblasts were weakly stimulated by recombinant (rec) tumor necrosis factor (rec TNF-alpha); they did, however, not proliferate in response to mitogens, lipopolysaccharide, rec granulocyte-macrophage colony stimulating factor (rec GM-CSF), macrophage-CSF, rec interferon-gamma, insulin or transferrin. The detection of IL 1 activity by crystal violet staining of human dermal fibroblasts was easier and faster than by measurement of thymidine incorporation of fibroblasts or mouse thymocytes; without loss of sensitivity, the sample capacity of the IL 1 assay could be enhanced, and the use of experimental animals was avoided.
Loppnow
H
H
Forschungsinstitut Borstel, Department of Immunology, Borstel, Federal Republic of Germany.
Flad
H D
HD
Dürrbaum
I
I
Musehold
J
J
Fetting
R
R
Ulmer
A J
AJ
Herzbeck
H
H
Brandt
E
E
D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007118:Immunoassay; D007375:Interleukin-1 / Q000032:analysis*; D008222:Lymphokines / Q000494:pharmacology; D012867:Skin / Q000378:metabolism*; D013194:Staining and Labeling; D013997:Time Factors
D007375:Interleukin-1; D008222:Lymphokines