| Autor: |
Mølvig, J., Sehested Hansen, B., Worsaae, H., HejnÆs, K. R., Helle, M., Dalbøge, H., Nerup, J. |
| Předmět: |
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| Zdroj: |
Scandinavian Journal of Immunology; Feb1990, Vol. 31 Issue 2, p225-235, 11p |
| Abstrakt: |
Recombinant human interleukin I β (rhIL- 1β) and supernatants of Escherichia coli lipopolysac- charides-stimulated human monocyte (Mo) cultures, containing native human IL- 1β (nhlL- 1β). demonstrate significant differences when tested in the mouse co-stimulatory thymocyte (lymphocyte activating Factor [LAF]) assay. The aims of the present study were to investigate this characteristic difference between rhlL- lβ and Moculture supernatants(Mo supernatants), and to compare the biological and the immunological activity of preparations of rhlL-1β and nhlL- 1β during each step of an identical purification procedure The biological activity of rhiL- lβ:nhIL- 1β preparations was characterized by the use of the LAY assay and the rat islet insulin release assay. An IL- 1β enzyme-linked immunosorbent assay (ELISA) was established in order to compare the biological and immunological responses of the IL- 1β preparations. We report that the significant difference between rh1L - 1β and supernatants of Mo cultures. which was only demonstrable in the LAF assay, is due to the presence of interleukin 6(IL-6) in the Mo supernatants. We describe a simple cation exchange chromatography separating nhIL.- 1β and 1L-6 of Mo supernatants. The highly purified rh1L- 1β possessing the correct amino-terminal sequence and nhIL- 1β have identical biological and immunological activities demonstrating a specific biologyica activity (SBA) of 3 × 102 using IL- 1β. Thus, we have no indications of secondary or tertiary structural differences between rhIL- 1β and purified nh1L-1β. In contrast, both in the LAF assay and in the rat islet insulin release assay the SBA of an amino-extended rh1L-1β from. Met-Glu-Ala-Glu-rh1L-1β was only 1.2% of the SBA of rh1L-1β suggesting that structural changes were introduced into the molecule by the amino-terminal extension. In the present study we have demonstrated that systematic combine testing of IL-1β preparations in two different biological assays and an immunological assay is useful for the characterization and comparison of the activity of recombinant and native IL-1β preparations purified by the use of exactly the same procedures. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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