title: Identification of the most active interleukin-32 isoform.
abstract: Cytokines are crucial in host defence against pathogens such as bacteria, viruses, fungi and parasites. A newly described cytokine, interleukin-32 (IL-32), induces various proinflammatory cytokines (tumour necrosis factor-alpha, IL-1beta, IL-6) and chemokines in both human and mouse cells through the nuclear factor-kappaB and p38 mitogen-activated protein kinase inflammatory signal pathway. The IL-32 primarily acts on monocytic cells rather than T cells. In an attempt to isolate the IL-32 soluble receptor, we used an IL-32 ligand-affinity column to purify neutrophil proteinase 3, which is a serine proteinase involved in many inflammatory diseases. IL-32 has biological activity associated with Mycobacterium tuberculosis and chronic proinflammatory diseases such as rheumatoid arthritis. IL-32 is transcribed as six alternative splice variants and the biological activity of each individual isoform remains unknown. Here, we cloned the complementary DNA of the four IL-32 isoforms (alpha, beta, gamma and delta) that are the most representative IL-32 transcripts. To produce recombinant protein with a high yield, the amino acids of two cysteine residues were mutated to serine residues, because serine residues are not conserved among different species. The multi-step purified recombinant IL-32 isoform proteins were assessed for their biological activities with different cytokine assays. The gamma isoform of IL-32 was the most active, although all isoforms were biologically active. The present study will provide a specific target to neutralize endogenous IL-32, which may contribute to basic and clinical immunology.
authors.affiliation: Laboratory of Cytokine Immunology, Institute of Biomedical Science and Technology, College of Medicine, Konkuk University, Seoul, Korea.
authors.forename: Ji-Da
authors.initials: JD
authors.lastname: Choi
authors.forename: Su-Young
authors.initials: SY
authors.lastname: Bae
authors.forename: Jae-Woo
authors.initials: JW
authors.lastname: Hong
authors.forename: Tania
authors.initials: T
authors.lastname: Azam
authors.forename: Charles A
authors.initials: CA
authors.lastname: Dinarello
authors.forename: Erk
authors.initials: E
authors.lastname: Her
authors.forename: Whan-Soo
authors.initials: WS
authors.lastname: Choi
authors.forename: Bo-Kyung
authors.initials: BK
authors.lastname: Kim
authors.forename: Chang-Kwon
authors.initials: CK
authors.lastname: Lee
authors.forename: Do-Young
authors.initials: DY
authors.lastname: Yoon
authors.forename: Sun-Jong
authors.initials: SJ
authors.lastname: Kim
authors.forename: Soo-Hyun
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D003545:Cysteine / Q000235:genetics; D016207:Cytokines / Q000096:biosynthesis; D004847:Epithelial Cells / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009154:Mutation; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D016207:Cytokines; C496971:IL32 protein, human; D007378:Interleukins; D020033:Protein Isoforms; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003545:Cysteine
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