Molecular Cloning and Expression of Rat Interleukin-1α cDNA

作者：Tsutomu Nishida, Naoki Nishino, Masaaki Takano, Yasuyo Sekiguchi, Kazuyoshi Kawai, Keiko Mizuno, Satoru Nakai, Yoshihiro Masui, Yoshikatsu Hirai · 发表于：The Journal of Biochemistry · 年份：1989 · DOI：10.1093/oxfordjournals.jbchem.a122667 · 被引用次数：327 · 研究领域：Immune Cell Function and Interaction、Immune Response and Inflammation、Toxin Mechanisms and Immunotoxins

A cDNA sequence coding for rat interleukin-1 alpha (IL-1 alpha) has been isolated from a cDNA library that was prepared with mRNA derived from LPS-stimulated rat peritoneal macrophages by using human IL-1 alpha cDNA as a probe. The rat cDNA encodes a 270 amino acid residue protein which is homologous (65%) to human IL-1 alpha. The rat cDNA sequence under SV40 early promoter directed the synthesis of biologically active IL-1 in monkey COS-1 cells. Rat IL-1 alpha mRNA is not expressed in spleen, lung, liver or brain, and is also not expressed in these organs of LPS-treated rat except spleen. This suggests that IL-1 alpha is not produced constitutively in various tissues and LPS is not sufficient to induce IL-1 alpha in most tissues. Our data indicate that the IL-1 activities which have been reported to be produced in the brain are not of alpha type. We have constructed a plasmid expressing the carboxy terminal 156 amino acids in Escherichia coli. Recombinant rat IL-1 alpha produced in COS cells or E. coli has cytotoxic activity against the human melanoma cell line A375S1 (GIF activity), which has been reported to be sensitive to human IL-1 alpha and IL-1 beta. This suggests that GIF activity is common to IL-1s derived from various sources.