Journal of Microbiology

Tumor Necrosis Factor Receptor (TNFR)-associated factor 2 (TRAF2) is not Involved in GM-CSF mRNA Induction and TNF-Mediated Cytotoxicity: Kim, Jung Hyun , Cha, Myung Hoon , Lee, Tae Kon , Seung, Hyo Jun , Park, Choon Sik , Chung, Il Yup; J. Microbiol. 1999;37(2):111-116.

Abstract: Tumor necrosis factor receptor (TNFR)-associated factor 2 (TRAF2) is known to act as a signal transducer that connects TNFR2 to its downstream effector functions such as proliferation of thymocytes, regulation of gene expression, and cell death. TRAF2 consists of largely two domains, the N-terminal half that contains a signal-emanating region and the C-terminal half that is responsible for binding to the intracellular region of TNFR2. In this study, we examined the possible roles of TRAF2 in granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression and cell death. A truncated mutant of TRAF2 (Δ2-263) that contains only a C-terminal half was generated, and transiently transfected to the A549 cell, a human lung cancer cell line, and L929 cell, a murine TNF-sensitive cell line. GM-CSF mRNA was induced in untransfected A540 cells both in dose- and time-dependent manner upon the exposure of TNF. However, neither the full length TRAF2 nor the mutant altered GM-CSF mRNA production regardless of the presence or absence of TNF. Furthermore, neither TRAF2 versions significantly changed the cytotoxic effect of TNF on L929 cells. These data suggest that TRAF2 may not be involved in the signal transduction pathway for GM-CSF gene induction and cell death mediated by TNF.

A Recombinant Mouse GM-CSF Protein Expressed as an Inclusion Form Shows Colony Stimulating Activity: Jin-Kyoo Kim , Eun-Jung Sohn , Soo-O Lee , Choon-Taek Lee , Ah Young Lee , Hye Kyung Chung , Bong Whan Sung , Hyun Joo Youn; J. Microbiol. 2000;38(2):109-112.

Abstract: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic hematopoietic growth factor and an activator of mature myeloid cells, and recombinant GM-CSF is increasingly under clinical studies for the treatment of various diseases including cancer, infectious diseases and hematopoietic diseases. We constructed a recombinant mouse GM-CSF expression plasmid with pelB leader sequence and His.Tag under T7 promoter control, and showed that the construct produced a 20 kDa recombinant protein in 8M urea. We also showed that the 20 kDa recombinant protein prepared in 8M urea stimulated colony formation in vitro, indicating that the recombinant mGM-CSF can be renatured to its native form to show the colony stimulating activity.

A Recombinant Human GM-CSF Protein Expressed as an Inclusion form in Escherichia coli Stimulates Colony Formation and Cell Proliferation in vitro: Ah Young Lee , Jin-Kyoo Kim , Hye Kyung Chung , Eun Kyong Bae , Jung Suk Hwang , Chung Won Cho , Dong Seok Lee , Jae Yong Han , Choon-Taek Lee , Soon-; J. Microbiol. 2002;40(1):77-81.

Abstract: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic hematopoietic growth factor involved in the development of myeloid cells from bone marrow, and an activator of mature myeloid cells functioning in a variety of antimicrobial and inflammatory responses. Recently, recombinant GM-CSF is increasingly under clinical study for treatment of various diseases including cancer, infectious diseases and hematopoietic diseases as well as for an immune response modulator. In this study, we constructed a recombinant human GM-CSF (rhGM-CSF) expression plasmid with a pelB leader sequence and His.Tag under T7 promoter control. The expression construct was shown to produce a recombinant protein of 20 kDa in the 8M urea preparation, indicating the rhGM-CSF may be expressed as an insoluble inclusion form. The 20 kDa recombinant protein in 8M urea was transformed into the water-soluble form by dialysis against PBS buffer (phosphate buffered saline). The soluble rhGM-CSF protein was shown to stimulate colony formation and cell proliferation in vitro, indicating that the rhGM-CSF could be refolded into its native form to show colony stimulating activity.

Search