Rabbit IL-1. Cloning, expression, biologic properties, and transcription during endotoxemia.
The cloning, sequencing, expression, and biologic activities of rabbit IL-1 alpha and beta are described. A cDNA library was constructed in lambda gt10 by using polyadenylated RNA extracted from rabbit adherent splenic macrophages 4 h after stimulation with endotoxin. By using the cDNA for human IL-1 beta and IL-1 alpha as hybridization probes, cDNA for both forms of rabbit IL-1 were isolated. The cDNA for rabbit IL-1 beta encodes a precursor polypeptide of 268 amino acids with an overall homology to human IL-1 beta of 74% (81% in the mature region coding for a 17.5 kDa carboxyl-terminal protein). The similarity between the two rabbit IL-1 forms is 31% for the entire molecule and 34% for the mature protein. The mature polypeptides of both forms were expressed in Escherichia coli. The recombinant proteins were purified to homogeneity and tested in a variety of biologic assays. Both forms produced typical endogenous pyrogen fevers in rabbits and augmented murine thymocyte and Th cell proliferation. Rabbit IL-1 alpha and beta were more pyrogenic in rabbits than human rIL-1 beta, whereas human rIL-1 alpha and beta were slightly more potent lymphocyte-activating factors. The recombinant rabbit proteins induced PGE and IL-1 production from human PBMC in vitro. A RIA for human IL-1 alpha did not recognize rabbit IL-1 alpha or beta, but rabbit IL-1 beta cross-reacted (as much as 30%) in a RIA for human IL-1 beta. Rabbits were injected with endotoxin and mRNA for both forms of IL-1 were observed primarily in the spleen and liver. The mRNA reached maximal levels after 60 min, then declined rapidly over the next 3 h, but were still present after 24 h. Liver tissue removed 4 h after endotoxin infusion produced lymphocyte-activating factors which were neutralized by more than 90% with a combination of goat anti-rabbit IL-1 alpha and anti-IL-1 beta.
Cannon
J G
JG
Department of Medicine, Tufts University, Boston, MA 02111.
Clark
B D
BD
Wingfield
P
P
Schmeissner
U
U
Losberger
C
C
Dinarello
C A
CA
Shaw
A R
AR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D005260:Female; D005334:Fever / Q000209:etiology; D007106:Immune Sera / Q000032:analysis; D007375:Interleukin-1 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D011458:Prostaglandins E / Q000096:biosynthesis; D011817:Rabbits; D012772:Shock, Septic / Q000209:etiology / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D007106:Immune Sera; D007375:Interleukin-1; D011458:Prostaglandins E; D004247:DNA