Interleukin-8: Baculovirus Expression and the Receptor Signal Transduction Pathway
dc.contributor.advisor | Stephen R. Grant | |
dc.contributor.committeeMember | Rafael Alvarez | |
dc.contributor.committeeMember | Paula Sumstom | |
dc.creator | Kang, Xiaoqiang | |
dc.date.accessioned | 2019-08-22T19:38:23Z | |
dc.date.available | 2019-08-22T19:38:23Z | |
dc.date.issued | 1994-06-01 | |
dc.date.submitted | 2013-09-13T08:54:07-07:00 | |
dc.description.abstract | Xiaoqiang, Kang., Interleukin-8: Baculovirus Expression and the Receptor Signal Transduction Pathway. Doctor of Philosophy (Biomedical Sciences), June, 1994, 150 pp., 4 tables, 36 illustrations, bibliography, 212 titles. The cDNA for human interleukin-8 (IL8) was subcloned from a bacterial source into the eukaryotic baculoviral vector expression system. Recombinant human IL-8 (rhIL-8) was synthesized and secreted from SF9 cells following infection of a recombinant virus harboring the full-length IL-8 structural gene. Recombinant human interleukin-8 was purified ([greater than] 600 fold) to homogeneity using preparative HPLC. The rhIL-8 preparation retained all of the physical, immunological, and biochemical properties of the natural product (monocyte-derived IL-8). Baculovirus vector expression coupled to preparative HPLC proved to be a very efficient method for large-scale recombinant interleukin production. Biochemical mechanisms that mediate IL-8 receptor-stimulated activities are poorly understood. In this study, I have explored the intracellular mechanism(s) induced by IL-8 in differentiated HL-60 cells. IL-8 induced a rapid and transient activation of phospholipase A2 in differentiated HL-60 cells. A consequence of phospholipase A2 activation was the release of arachidonic acid and the generation of lysophospholipids from membrane phospholipids. The IL-8 stimulated-arachidonic acid release was pertussis toxin and phospholipase A2 inhibitor sensitive, and protein kinase C independent. In contrast to another neutrophil chemotactic factor, fMLP, IL-8 did not stimulate the activation of phospholipase C and phospholipase D. When comparing the phosphorylation events induced by IL-8 and fMLP, I found that these two chemotactic factors triggered different protein phosphorylation profiles. Tyrosine phosphorylation of proteins was not detected following IL-8 stimulation in HL-60 cells. However, IL-8 stimulated the rapid autophosphorylation of calcium/calmodulin-dependent protein kinase II (CaM kinase II). These results strongly suggest that the IL-8 receptor is closely coupled to the activation of PLA2 and that CaM kinase II is an integral component of IL-8 receptor signal pathway. | |
dc.format.mimetype | application/pdf | |
dc.identifier.uri | https://hdl.handle.net/20.500.12503/26536 | |
dc.language.iso | en | |
dc.provenance.legacyDownloads | 0 | |
dc.subject | Cell and Developmental Biology | |
dc.subject | Cell Biology | |
dc.subject | Cells | |
dc.subject | Immunology and Infectious Disease | |
dc.subject | Infectious Disease | |
dc.subject | Life Sciences | |
dc.subject | Medical Cell Biology | |
dc.subject | Medical Microbiology | |
dc.subject | Medicine and Health Sciences | |
dc.subject | Microbiology | |
dc.subject | Other Cell and Developmental Biology | |
dc.subject | Other Immunology and Infectious Disease | |
dc.subject | Virology | |
dc.subject | Virus Diseases | |
dc.subject | Viruses | |
dc.subject | Recombinant human interleukin-8 | |
dc.subject | eukaryotic baculoviral vector expression | |
dc.subject | PLA2 | |
dc.subject | CaM kinase II | |
dc.subject | calcium/calmodulin-dependent protein kinase II | |
dc.subject | autophosphorylation | |
dc.subject | baculovirus | |
dc.title | Interleukin-8: Baculovirus Expression and the Receptor Signal Transduction Pathway | |
dc.type | Dissertation | |
dc.type.material | text | |
thesis.degree.department | Graduate School of Biomedical Sciences | |
thesis.degree.discipline | Biomedical Sciences | |
thesis.degree.grantor | University of North Texas Health Science Center at Fort Worth | |
thesis.degree.name | Doctor of Philosophy |
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