Development of a simple, sensitive and specific bioassay for interleukin-1 based on the proliferation of RPMI 1788 cells. Comparison with other bioassays for IL-1.
The IL-1-dependent proliferation of RPMI 1788, a human EBV-transformed cell line, was used to develop a biological assay system for IL-1. Preparations of rhIL-1 alpha and rhIL-1 beta, as well as rmIL-1 beta exhibited a specific biological activity (50% of the maximal response) between 5.8 x 10(8) and 8.6 x 10(8) U/mg. Remarkably, a 3-5-fold reduced specific biological activity was noticed for rm-IL-1 alpha, viz. 1.7 x 10(8) U/mg. The IL-1-dependent proliferation of RPMI 1788 cells was compared with other IL-1 test systems, such as the IL-1-mediated induction of IL-2 in EL4-NOB-1, LBRM-33-1A5 and thymocytes, and the IL-1-driven induction of cytotoxic activity by PC60 cells, the so-called CIA assay. The cytokine-dependent growth of RPMI 1788 cells is highly specific for IL-1, and no other cytokine tested induced a proliferative response. The presence of high concentrations of rmTNF, rhTNF or rhIL-6 did not interfere with the quantification of IL-1. Additionally, we evaluated the detection of IL-1 in the presence of mitogens, phorbol ester or calcium ionophore, as well as the determination of IL-1 in serum and PF samples of human and murine origin.
Vandenabeele
P
P
Laboratory of Molecular Biology, State University of Ghent, Belgium.
Declercq
W
W
Libert
C
C
Fiers
W
W
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects; D001681:Biological Assay / Q000379:methods*; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis* / Q000097:blood; D007476:Ionophores / Q000494:pharmacology; D051379:Mice; D008809:Mice, Inbred C3H; D008934:Mitogens / Q000494:pharmacology; D010529:Peritoneal Cavity; D010703:Phorbol Esters / Q000494:pharmacology; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects
D007375:Interleukin-1; D007476:Ionophores; D008934:Mitogens; D010703:Phorbol Esters; D011994:Recombinant Proteins