Purification and biological function study of interleukin-22 Purification and characterization of novel protein of SARS and generation of monoclonal antibody
| Autor: | Chia -ying Yen, 顏佳瑩 |
|---|---|
| Rok vydání: | 2004 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 92 Interleukin-10 is a pleiotropic immunosuppressive and immune-stimulatory cytokine. IL-22 belongs to the interleukin-10 (IL-10)family, which includes IL-10, IL-19, IL-20, IL-22, MDA-7(IL-24), and AK155 (IL-26). Interleukin-22 encodes a protein of 179 amino acids that shares 25%identify with interleukin-10, and structural similarity with IL-10. The biological activities of IL-22 known thus for are the induction of acute-phase reactants in liver and hepatoma cells as well as induction of pancreatitis-associated protein (PAP1) in pancreatic acinar cells, suggesting its role in inflammatory response .To further explore any novel biological fuction of IL-22 and the interaction of IL-22 with other cytokine and anelyze it’s biological function on monocye; we purified IL-22 recombinant proteins expressed in the yeast pichia system. I treated monocytes with IL-22 and analyzed the induced cytokines. To explore if IL-22 was also associated with pathogenesis of asthma and pancreatic cancer, we employed ELISA to analyze the serum level of IL-22 in the patients. The results demonstrated that treatment of monocytes with IL-22 induced dose-dependent production of IL-6 and TNF-α. To further study the relationships between IL-22 and other members of IL-10 family; IL-10 and IL-19, monocyte were treated with different combination of IL-10, IL-19 and IL-22. IL-22 induced the production of IL-6 and TNF-α,which was slightly enhanced by the addition of IL-19. IL-10 completely inhibited the induction of IL-6 and TNF-αby IL-22. There is no significant difference in the IL-22 serum level of asthmatic and pancreatic cancer patients compared to the normal control. Severe Acute Respiratory Syndrome (SARS) is a disease caused by a coronavirus. The genome of SARS-CoV containing 29,727 nucleotides in length, has 11 open reading frames. The genome organization is similar to that of other corona-viruses. Phylogenetic analyses and sequence comparison showed that SARS-CoV is not closely related to any of the previously characterized corona-viruses. The genome of SARS-CoV has several novel open reading frames that could be of biological significance. SARS patients have been shown to have serious infiltrate of lymphocytes in the lung and high level of inflammatory cytokines such as IL-6, IL-8 and TNF-alpha. My study was aimed to study the functions of ORF-10 protein encoded by open reading frames from SARS virus genome. To this aim I constructed and expressed the ORF-10 protein in vitro in Pichia; E.coli and mammalian cells. The recombinant protein will be analyzed in vitro such as lung cells or immune cells to see if it induces any chemokine or cytokine. I also generated the monoclonal antibody against this protein to explore its therapeutic potential. I also demonstrated that ORF10 is a cytosol protein . |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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