Purification and characterization of the immunostimulatory properties of recombinant human interleukin-1 beta.
In this study we have used a new method for human recombinant IL-1 beta (rIL-1 beta) purification and investigated its immunostimulatory biological activity. The IL-1 beta gene was cloned using a novel mRNA preparation from activated human blood monocytes. The purification protocol consists of extraction and two chromatographic steps using the new Soloza cation exchange resin. The purified protein was characterized electrophoretically, by amino acid analysis and reverse phase chromatography. The protein migrated on SDS-PAGE with a molecular weight of 18.200 but demonstrated the minor presence of aggregates (dimers and trimers). Specific activity of purified rIL-1 beta in comitogenic assay on mouse thymocytes was 10(8) U/mg protein. rIL-1 beta increased in a dose dependent manner proliferation of Con A-stimulated murine thymocytes, splenocytes, PHA-stimulated human peripheral blood lymphocytes and transformed B-cell lines. Comitogenic activity depended on the degree of lymphocyte preactivation and was similar to that of natural human IL-1 beta. rIL-1 beta enhanced IL-2 production by murine spleen cells and EL-4 cell line and IL-2 receptor expression by human peripheral blood mononuclear cells. It induced PGE2 release from human blood monocytes but had no effect on human neutrophil chemotaxis, phagocytosis and respiratory burst.
Ketlinsky
S
S
All-Union Research Institute of Highly Pure Biopreparations, Leningrad, USSR.
Simbirtsev
A
A
Poltorack
A
A
Protasov
E
E
Solovjeva
L
L
Putchkova
G
G
Konusova
V
V
Pigareva
N
N
Kalinina
N
N
Perumov
N D
ND
D000818:Animals; D002852:Chromatography, Ion Exchange; D015232:Dinoprostone / Q000096:biosynthesis; D006801:Humans; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D007376:Interleukin-2 / Q000096:biosynthesis; D008213:Lymphocyte Activation; D051379:Mice; D008934:Mitogens; D008970:Molecular Weight; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology
D007375:Interleukin-1; D007376:Interleukin-2; D008934:Mitogens; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D015232:Dinoprostone