PRODUCTION OF LARGE AMOUNTS OF RECOMBINANT INTERLEUKINS BY CDNA TRANSFECTED MOUSE MYELOMA CELLS CULTURED IN DIALYSIS TUBING

Studies of interleukin function often require large quantities of thes e highly expensive substances. The available interleukins are generall y recombinant proteins produced in bacteria or yeast and, less commonl y, interleukins produced by mammalian cells, which provide appropriate glycosylation and other post-translational modifications. Due to diff erences in biosynthesis, difficulties in production and purification t he quality of the interleukin preparations may vary. We have taken adv antage of the recently developed constitutively interleukin-secreting mouse myeloma cell lines and the dialysis tubing culture technique, wh ich permit cells to be grown at high densities, inorder to establish a method for the production of large amounts of recombinant murine IL-2 and IL-4. We show that these interleukins can be produced at low cost and in concentrations 20-30-fold higher than in conventional culture flasks. A single dialysis tubing culture will produce more than 10(6) U of interleukin which may be compared with the available commercial p reparations containing between 10- and a 100-fold less per vial. The I L-2 and IL-4 produced in this manner are biologically active molecules as demonstrated by the strong proliferative response of clonal T cell s and the isotype-switching effect in LPS-stimulated splenic B cell cu ltures. The dialysis tubing culture technique is a simple and highly c ost-effective means of generating large quantities of biologically act ive interleukins and is especially suitable for research laboratories interested in functional studies of these proteins.