IL-24; a glycosylated dimeric cytokine affecting monocytes and cytotoxic to melanoma cells | Posted on:2006-08-18 | Degree:Ph.D | Type:Dissertation | University:The University of Texas Graduate School of Biomedical Sciences at Houston | Candidate:Mumm, John | Full Text:PDF | GTID:1454390008454984 | Subject:Health Sciences | Abstract/Summary: | | IL-24 is an unusual member of the IL-10 family, which is considered a Th1 cytokine that exhibits tumor cell cytotoxicity. I describe the purification of this novel cytokine from the supernatant of IL-24 gene transfected human embryonic kidney cells and define the biochemical and functional properties of the soluble, human IL-24 protein.; I showed IL-24 non-covalently associates with bovine albumin. Immunoaffinity purification followed by cation exchange chromatography resulted in the significant enrichment of N-glycosylated IL-24. This protein elicited dose-dependent secretion of TNF-alpha and IL-6 from purified human monocytes and TNF-alpha secretion from PMA differentiated U937 cells. I showed this same protein was cytotoxic to melanoma tumor cells via the induction of IFN-alpha.; I reported IL-24 associates as at least two disulfide linked, N-glycosylated dimers. Enzymatic removal of N-linked-glycosylation from purified IL-24 partially diminished its cytokine and cytotoxic functions. Disruption of IL-24 dimers via reduction and alkylation of intermolecular disulfide bonds nearly abolished IL-24s cytokine function.; I elucidated IL-24 induced TNF-alpha secretion was pSTAT1, pSTAT3 as well as the class II heterodimeric receptors IL-20R1/IL-22R2 independent. I identified a requirement for the heterodimer of Toll-like Receptors 1 and 2 for IL-24s cytokine function and show a physical interaction between IL-24 and the extracellular domain of TLR-1.; Thus, I demonstrated that purified N-glycosylated, soluble, dimeric, human IL-24 exhibits both immunomodulatory and anti-cancer activities and these functions remain associated during purification. IL-24 induced TNF-alpha secretion required an interaction with the heterodimeric receptor TLR-1/2 and IL-24s cytotoxic affect to melanoma tumor cells was in part due to its induction of IFN-beta. | Keywords/Search Tags: | Cytokine, Cytotoxic, IL-24 induced tnf-alpha secretion, Melanoma, Human IL-24 | | Related items |
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