Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells
碩士 === 國立臺灣大學 === 園藝學研究所 === 93 === Abstract IPAF (immunomodulatory protein from Anoectochilus formosanus Hayata) was purified from Anoectochilus formosanus Hayata by 30% saturation of ammonium sulfate precipitation. SDS-PAGE analysis showed that IPAF has a molecular weight of 14 kDa and isoelectr...
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ndltd-TW-093NTU053790452015-10-13T11:12:49Z http://ndltd.ncl.edu.tw/handle/06377654483210925949 Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells 台灣金線連免疫調節蛋白IPAF活化免疫細胞之機制 Ju-Chun Hsu 許如君 碩士 國立臺灣大學 園藝學研究所 93 Abstract IPAF (immunomodulatory protein from Anoectochilus formosanus Hayata) was purified from Anoectochilus formosanus Hayata by 30% saturation of ammonium sulfate precipitation. SDS-PAGE analysis showed that IPAF has a molecular weight of 14 kDa and isoelectric focusing electrophoresis revealed the isoelectric point of IPAF was 5.4. Culturing RAW 264.7 macrophages with IPAF in vitro showed that IPAF could activate the cells and then increase nitric oxide, TNF-α and IL-1β production. Moreover, IPAF also stimulated the proliferation of murine splenocytes and increased their IFN-γ secretion, but did not result in IL-4 production. However, the cell depletion test demonstrated that IPAF increased the B cell proliferation without T cells existing. These results suggested that IPAF was a thymus-independent antigen. Peritoneal cells from C57BL/10ScN mice, which have a null mutation in TLR4 (TLR4-/-), were hyporesponsive in TNF-α secretion to both LPS and IPAF, suggesting that LPS and IPAF may share a same signaling pathway involving TLR4. On the other hand, IPAF could increase TLR4-/- B cells proliferation. Therefore, we propose that IPAF might be not involved in TLR4 signaling pathway and have another signaling pathway in B cell proliferation. Taking together, this study clearly demonstrated that IPAF, which activated B cells and also activated macrophages through TLR4, is an immune stimulus and could be helpful to strength the immunity of its host. 許輔 2005 學位論文 ; thesis 138 zh-TW |
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碩士 === 國立臺灣大學 === 園藝學研究所 === 93 === Abstract
IPAF (immunomodulatory protein from Anoectochilus formosanus Hayata) was purified from Anoectochilus formosanus Hayata by 30% saturation of ammonium sulfate precipitation. SDS-PAGE analysis showed that IPAF has a molecular weight of 14 kDa and isoelectric focusing electrophoresis revealed the isoelectric point of IPAF was 5.4. Culturing RAW 264.7 macrophages with IPAF in vitro showed that IPAF could activate the cells and then increase nitric oxide, TNF-α and IL-1β production. Moreover, IPAF also stimulated the proliferation of murine splenocytes and increased their IFN-γ secretion, but did not result in IL-4 production. However, the cell depletion test demonstrated that IPAF increased the B cell proliferation without T cells existing. These results suggested that IPAF was a thymus-independent antigen.
Peritoneal cells from C57BL/10ScN mice, which have a null mutation in TLR4 (TLR4-/-), were hyporesponsive in TNF-α secretion to both LPS and IPAF, suggesting that LPS and IPAF may share a same signaling pathway involving TLR4. On the other hand, IPAF could increase TLR4-/- B cells proliferation. Therefore, we propose that IPAF might be not involved in TLR4 signaling pathway and have another signaling pathway in B cell proliferation.
Taking together, this study clearly demonstrated that IPAF, which activated B cells and also activated macrophages through TLR4, is an immune stimulus and could be helpful to strength the immunity of its host.
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許輔 |
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許輔 Ju-Chun Hsu 許如君 |
author |
Ju-Chun Hsu 許如君 |
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Ju-Chun Hsu 許如君 Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
author_sort |
Ju-Chun Hsu |
title |
Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
title_short |
Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
title_full |
Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
title_fullStr |
Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
title_full_unstemmed |
Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells |
title_sort |
activating mechanism of immunomodulatory protein from anoectochius formosanus (ipaf) on immune cells |
publishDate |
2005 |
url |
http://ndltd.ncl.edu.tw/handle/06377654483210925949 |
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