Evaluation of the Immunoregulatory Activity of rFip-gts Purified from Baculovirus-infected Insect Cells
Fip-gts, an immunomodulatory protein purified from Ganoderma tsugae, has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. For medicinal application, a recombinant Fip-gts was successfully expressed and purified in Sf21 insect cells by our previously work. It is important to evaluate the immunomodulatory activity of the rFip-gts. To assess the immunomodulatory potential of rFip-gts, the T lymphocytes of murine splenocytes were used in the present study. Results revealed that rFip-gts induced cellular aggregation formation. Additionally, the expression of IL-2 and IFN-r were up-regulated after the treatment of rFip-gts, and a corresponding increased production of IL-2 and IFN-r in a dose-dependent manner. The results showed that rFip-gts has an immunomodulatory activity in inducing Th1 lymphocytes from murine splenocytes released IL-2 and IFN-γ, thus suggest that rFip-gts may have therapeutic potential in vivo as an immune modulator.
[1] Chiang Su New Medical College, "Dictionary of Chinese crude drug
Shanghai Scientific Technological Publishers," Shanghai, Chinese, 1979,
pp.16.
[2] W. H. Lin, C. H. Hung, C. I. Hsu, and J.Y. Lin, "Dimerization of the
N-terminal amphipathic alpha-helix domain of the fungal
immunomodulatory protein from Ganoderma tsugae (Fip-gts) defined by a
yeast two-hybrid system and site-directed mutagenesis," J Biol Chem,
vol.272, pp. 20044-20048, Aug. 1997.
[3] S. Tanaka, K. Ko, K. Kino, K. Tsuchiya, A. Yamashita, A. Murasugi, S.
Sakuma, and H. Tsunoo, "Complete amino acid sequence of an
immunomodulatory protein, ling zhi-8 (LZ-8)," J Biol Chem, vol. 264,
pp.16372-16377, Oct. 1989.
[4] K. Kino, A. Yamashita, K. Yamaoka, J. Watanabe, S. Tanaka, K. Ko, K.
Shimizu, and H. Tsunoo, "Isolation and characterization of a new
immunomodulatory protein, ling zhi-8 (LZ-8), from Ganoderma lucidium,"
J Biol Chem, vol. 264, pp. 472-478, Jan. 1989.
[5] N. S. Lai, R. H. Lin, R. S. Lai, U. C. Kun, S. C. Leu, "Prevention of
autoantibody tormation and prolonged survival in New Zealand Black/New
Zealand white F1 mice with anancient hinese herb, Ganoderma tsugae,"
Lupus., vol. 10, pp. 461-465, 2001
[6] C. H. Liao, Y. M. Hsiao, C. P. Hsu, M. Y. Lin, J. C. Wang, Y. L. Huang, and
J. L. Ko, "Transcriptionally mediated inhibition of telomerase of fungal
immunomodulatory protein from Ganoderma tsugae in A549 human lung
adenocarcinoma cell line," Mol. Carcinog., vol. 45, pp. 220-229, Apr.
2006.
[7] C. H. Liao, Y. M. Hsiao, C. H. Lin, C. S. Yeh, J. C. Wang, C. H. Ni, C. P.
Hsu, and J. L. Ko, "Induction of premature senescence in human lung
cancer by fungal immunomodulatory protein from Ganoderma tsugae,"
Food. Chem. Toxicol., vol. 46, pp. 1851-1859, May. 2008.
[8] P. H. Wang, S. F. Yang, G. D. Chen, C. P. Han, S. C. Chen, L. Y. Lin, and J.
L .Ko, "Human nonmetastatic clone 23 type 1 gene suppress migration of
cervical cancer cells and enhances the migration inhibition of fungal
immunomodulatory protein from Ganoderma tsugae," Reprod. Sci., vol. 14,
pp. 475-485, Jul. 2009.
[9] T. R .Jinn, C. M. Wu, W. C. Tu, J. L. Ko, and Jason T. C. Tzen "Functional
expression of Fip-gts, a fungal immunomodulatory protein from
Ganoderma tsugae in Sf21 insect cells," Biosci. Biotechnol. Biochem., vol.
70, pp. 2627-2634, Nov. 2006.
[10] T. Y. Wu, C. M. Wu, J. L. Ko, T .R. Jinn, "Efficient production of the
immunomodulatory glycoprotein Fip-gts from Ganoderma tsugae in
baculovirus-infected insect larvae," Biosci. Biotechnol. Biochem.,
submitted for publication.
[11] C. L. C. Giraudo, L. Rumi, and C. D. Pasqualini, "An improved method for
PHA-culturing mouse lymphocytes using Ficoll-Hypaque gradient," Boll.
Ist. Sieroter. Milan, vol. 55, pp.304-307, Sep. 1976.
[12] A. S. Wermerskirchen, D. H. LaTocha, and B. L. Clarke,
"Adrenocorticotropic hormone controls Concanavalin A activation of rat
lymphocytes by modulating IL-2 production," Life Science, vol. 67, pp.
2177-2187, Sep. 2000.
[1] Chiang Su New Medical College, "Dictionary of Chinese crude drug
Shanghai Scientific Technological Publishers," Shanghai, Chinese, 1979,
pp.16.
[2] W. H. Lin, C. H. Hung, C. I. Hsu, and J.Y. Lin, "Dimerization of the
N-terminal amphipathic alpha-helix domain of the fungal
immunomodulatory protein from Ganoderma tsugae (Fip-gts) defined by a
yeast two-hybrid system and site-directed mutagenesis," J Biol Chem,
vol.272, pp. 20044-20048, Aug. 1997.
[3] S. Tanaka, K. Ko, K. Kino, K. Tsuchiya, A. Yamashita, A. Murasugi, S.
Sakuma, and H. Tsunoo, "Complete amino acid sequence of an
immunomodulatory protein, ling zhi-8 (LZ-8)," J Biol Chem, vol. 264,
pp.16372-16377, Oct. 1989.
[4] K. Kino, A. Yamashita, K. Yamaoka, J. Watanabe, S. Tanaka, K. Ko, K.
Shimizu, and H. Tsunoo, "Isolation and characterization of a new
immunomodulatory protein, ling zhi-8 (LZ-8), from Ganoderma lucidium,"
J Biol Chem, vol. 264, pp. 472-478, Jan. 1989.
[5] N. S. Lai, R. H. Lin, R. S. Lai, U. C. Kun, S. C. Leu, "Prevention of
autoantibody tormation and prolonged survival in New Zealand Black/New
Zealand white F1 mice with anancient hinese herb, Ganoderma tsugae,"
Lupus., vol. 10, pp. 461-465, 2001
[6] C. H. Liao, Y. M. Hsiao, C. P. Hsu, M. Y. Lin, J. C. Wang, Y. L. Huang, and
J. L. Ko, "Transcriptionally mediated inhibition of telomerase of fungal
immunomodulatory protein from Ganoderma tsugae in A549 human lung
adenocarcinoma cell line," Mol. Carcinog., vol. 45, pp. 220-229, Apr.
2006.
[7] C. H. Liao, Y. M. Hsiao, C. H. Lin, C. S. Yeh, J. C. Wang, C. H. Ni, C. P.
Hsu, and J. L. Ko, "Induction of premature senescence in human lung
cancer by fungal immunomodulatory protein from Ganoderma tsugae,"
Food. Chem. Toxicol., vol. 46, pp. 1851-1859, May. 2008.
[8] P. H. Wang, S. F. Yang, G. D. Chen, C. P. Han, S. C. Chen, L. Y. Lin, and J.
L .Ko, "Human nonmetastatic clone 23 type 1 gene suppress migration of
cervical cancer cells and enhances the migration inhibition of fungal
immunomodulatory protein from Ganoderma tsugae," Reprod. Sci., vol. 14,
pp. 475-485, Jul. 2009.
[9] T. R .Jinn, C. M. Wu, W. C. Tu, J. L. Ko, and Jason T. C. Tzen "Functional
expression of Fip-gts, a fungal immunomodulatory protein from
Ganoderma tsugae in Sf21 insect cells," Biosci. Biotechnol. Biochem., vol.
70, pp. 2627-2634, Nov. 2006.
[10] T. Y. Wu, C. M. Wu, J. L. Ko, T .R. Jinn, "Efficient production of the
immunomodulatory glycoprotein Fip-gts from Ganoderma tsugae in
baculovirus-infected insect larvae," Biosci. Biotechnol. Biochem.,
submitted for publication.
[11] C. L. C. Giraudo, L. Rumi, and C. D. Pasqualini, "An improved method for
PHA-culturing mouse lymphocytes using Ficoll-Hypaque gradient," Boll.
Ist. Sieroter. Milan, vol. 55, pp.304-307, Sep. 1976.
[12] A. S. Wermerskirchen, D. H. LaTocha, and B. L. Clarke,
"Adrenocorticotropic hormone controls Concanavalin A activation of rat
lymphocytes by modulating IL-2 production," Life Science, vol. 67, pp.
2177-2187, Sep. 2000.
@article{"International Journal of Medical, Medicine and Health Sciences:56003", author = "Tzong Yuan Wu and Sheng Kuo Hsieh and Tzyy Rong Jinn", title = "Evaluation of the Immunoregulatory Activity of rFip-gts Purified from Baculovirus-infected Insect Cells", abstract = "Fip-gts, an immunomodulatory protein purified from Ganoderma tsugae, has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. For medicinal application, a recombinant Fip-gts was successfully expressed and purified in Sf21 insect cells by our previously work. It is important to evaluate the immunomodulatory activity of the rFip-gts. To assess the immunomodulatory potential of rFip-gts, the T lymphocytes of murine splenocytes were used in the present study. Results revealed that rFip-gts induced cellular aggregation formation. Additionally, the expression of IL-2 and IFN-r were up-regulated after the treatment of rFip-gts, and a corresponding increased production of IL-2 and IFN-r in a dose-dependent manner. The results showed that rFip-gts has an immunomodulatory activity in inducing Th1 lymphocytes from murine splenocytes released IL-2 and IFN-γ, thus suggest that rFip-gts may have therapeutic potential in vivo as an immune modulator.
", keywords = "Fungal immunomodulatory protein, Ganodermatsugae, Interleukin 2, Interferon γ, Lingzhi.", volume = "4", number = "5", pages = "205-3", }
