Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms
Aktivni sastojci gljiva - ispitivanje promene imunomodulatorske aktivnosti enzimski modifikovanih glukana gljiva iz klase Basidiomycetes
Апстракт
Chemical modifications of bioactive β-D-glucans in higher mushrooms are often necessary to improve their immunomodulating properties and clinical qualities. The aim of the investigation was to examine immunomodulating activity of native myco-glucans and glucans of lower molecular weight (MW lt 80kDa) remaining after enzymatic degradation with 1,6 β-glucanase and to evaluate structural features of primary stimulation of human immune cells. The immunomodulating activity of native and enzimatically modified glucans of Agaricus blazei and Agaricus bisporus fruit body powder was tested in vitro by stimulation peripheral blood mononuclear cells (PBMC's) and measuring interferon-gamma (IFN-γ) production by enzyme linked immunosorbent assay (ELISA). Titers of IFN-γ measured after incubation of activated cells with native A. blazei and A. bisporus glucans confirmed their characteristic immunostimulating activity on PBMC's. For A. bisporus the resulting β-(1,3)-glucan fragments of high molecular... weight (MW>80 kDa) and the small fragments (MW lt 80 kDa) of A. blazei β-(1,3)-glucan remaining after enzymatic modification showed a significant enhancement of immunostimulating activity compared to the native glucans. The results confirmed that β-(1,3) bonds are the primary determinants of immunomodulatory activities and stimulation of IFN-γ synthesis. It indicated that primary structure is more important than the tertiary structure of the triple helix for their immunostimulating activity and synthesis of IFN-γ. The results confirmed that glucans of lower molecular weight are effective stimulatiors of IFN-γ synthesis.
Hemijske modifikacije biološki aktivnih β-D-glukana viših gljiva su često neophodne kako bi se poboljšala njihova imunomodulatorska svojstva, a samim tim i njihova klinička primena. Cilj rada je ispitivanje imunomodulatorske aktivnosti nativnih miko-glukana, kao i glukana manjih molekulskih masa (MM lt 80 KD), nastalih nakon enzimske modifikacije 1,6 β-glukanazama i utvrđivanje koja je strukturna karakteristika primarno značajna za stimulaciju ćelija humanog imunog sistema. Imunomodulatorska aktivnost nativnih i enzimski modifikovanih glukana izolovanih iz praha plodonosnih tela gljiva Agaricus blazei i Agaricus bisporus testirana je in vitro stimulacijom mononuklearnih ćelija krvi iz perifernog krvotoka (PBMCPeripheral Blood Mononuclear Cell) molekulima glukana i merenjem količine sintetisanog interferona-gama (IFN-γ) od strane stimulisanih ćelija enzimoimunotestom (ELISA-enzyme linked immunosorbent assay). Merenjem titra IFN-γ nastalog nakon inkubacije aktiviranih ćelija sa nativnim ...A. blazei i A. bisporus glukanima potvrđeno je da su ovi molekuli ispoljili izraženo imunostimulativno dejstvo na PBMC ćelije. A. bisporus β-(1,3)-glukani velikih molekulskih masa (MM>80 kDa), kao i mali fragmenti A. blazei β-(1,3)-glukana (MM lt 80 kDa), nastali nakon enzimske modifikacije, ispoljili su značajno povećanje imunostimulativne aktivnosti u odnosu na nativne molekule. Ovim je potvrđeno da je za imunostimulativnu aktivnost i stimulaciju sinteze IFN-γ primarno značajno prisustvo β-(1,3)-glikozidnih veza, odnosno da je bitna primarna struktura, a ne konformacija trostrukog heliksa nativnih molekula. Dobijenim rezultatima je pokazano da su glukani manjih molekulskih masa efikasni stimulatori sinteze IFN-γ.
Кључне речи:
Agaricus blazei / Agaricus bisporus / glucans / immunomodulating activity / IFN-γ / Agaricus blazei / Agaricus bisporus / glukani / imunomodulatorskaaktivnost / IFN-γИзвор:
Hrana i ishrana, 2010, 51, 1-2, 15-20Издавач:
- Društvo za ishranu Srbije, Beograd
Институција/група
Poljoprivredni fakultetTY - JOUR AU - Kozarski, Maja AU - Klaus, Anita AU - Nikšić, Miomir PY - 2010 UR - http://aspace.agrif.bg.ac.rs/handle/123456789/2185 AB - Chemical modifications of bioactive β-D-glucans in higher mushrooms are often necessary to improve their immunomodulating properties and clinical qualities. The aim of the investigation was to examine immunomodulating activity of native myco-glucans and glucans of lower molecular weight (MW lt 80kDa) remaining after enzymatic degradation with 1,6 β-glucanase and to evaluate structural features of primary stimulation of human immune cells. The immunomodulating activity of native and enzimatically modified glucans of Agaricus blazei and Agaricus bisporus fruit body powder was tested in vitro by stimulation peripheral blood mononuclear cells (PBMC's) and measuring interferon-gamma (IFN-γ) production by enzyme linked immunosorbent assay (ELISA). Titers of IFN-γ measured after incubation of activated cells with native A. blazei and A. bisporus glucans confirmed their characteristic immunostimulating activity on PBMC's. For A. bisporus the resulting β-(1,3)-glucan fragments of high molecular weight (MW>80 kDa) and the small fragments (MW lt 80 kDa) of A. blazei β-(1,3)-glucan remaining after enzymatic modification showed a significant enhancement of immunostimulating activity compared to the native glucans. The results confirmed that β-(1,3) bonds are the primary determinants of immunomodulatory activities and stimulation of IFN-γ synthesis. It indicated that primary structure is more important than the tertiary structure of the triple helix for their immunostimulating activity and synthesis of IFN-γ. The results confirmed that glucans of lower molecular weight are effective stimulatiors of IFN-γ synthesis. AB - Hemijske modifikacije biološki aktivnih β-D-glukana viših gljiva su često neophodne kako bi se poboljšala njihova imunomodulatorska svojstva, a samim tim i njihova klinička primena. Cilj rada je ispitivanje imunomodulatorske aktivnosti nativnih miko-glukana, kao i glukana manjih molekulskih masa (MM lt 80 KD), nastalih nakon enzimske modifikacije 1,6 β-glukanazama i utvrđivanje koja je strukturna karakteristika primarno značajna za stimulaciju ćelija humanog imunog sistema. Imunomodulatorska aktivnost nativnih i enzimski modifikovanih glukana izolovanih iz praha plodonosnih tela gljiva Agaricus blazei i Agaricus bisporus testirana je in vitro stimulacijom mononuklearnih ćelija krvi iz perifernog krvotoka (PBMCPeripheral Blood Mononuclear Cell) molekulima glukana i merenjem količine sintetisanog interferona-gama (IFN-γ) od strane stimulisanih ćelija enzimoimunotestom (ELISA-enzyme linked immunosorbent assay). Merenjem titra IFN-γ nastalog nakon inkubacije aktiviranih ćelija sa nativnim A. blazei i A. bisporus glukanima potvrđeno je da su ovi molekuli ispoljili izraženo imunostimulativno dejstvo na PBMC ćelije. A. bisporus β-(1,3)-glukani velikih molekulskih masa (MM>80 kDa), kao i mali fragmenti A. blazei β-(1,3)-glukana (MM lt 80 kDa), nastali nakon enzimske modifikacije, ispoljili su značajno povećanje imunostimulativne aktivnosti u odnosu na nativne molekule. Ovim je potvrđeno da je za imunostimulativnu aktivnost i stimulaciju sinteze IFN-γ primarno značajno prisustvo β-(1,3)-glikozidnih veza, odnosno da je bitna primarna struktura, a ne konformacija trostrukog heliksa nativnih molekula. Dobijenim rezultatima je pokazano da su glukani manjih molekulskih masa efikasni stimulatori sinteze IFN-γ. PB - Društvo za ishranu Srbije, Beograd T2 - Hrana i ishrana T1 - Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms T1 - Aktivni sastojci gljiva - ispitivanje promene imunomodulatorske aktivnosti enzimski modifikovanih glukana gljiva iz klase Basidiomycetes EP - 20 IS - 1-2 SP - 15 VL - 51 UR - https://hdl.handle.net/21.15107/rcub_agrospace_2185 ER -
@article{ author = "Kozarski, Maja and Klaus, Anita and Nikšić, Miomir", year = "2010", abstract = "Chemical modifications of bioactive β-D-glucans in higher mushrooms are often necessary to improve their immunomodulating properties and clinical qualities. The aim of the investigation was to examine immunomodulating activity of native myco-glucans and glucans of lower molecular weight (MW lt 80kDa) remaining after enzymatic degradation with 1,6 β-glucanase and to evaluate structural features of primary stimulation of human immune cells. The immunomodulating activity of native and enzimatically modified glucans of Agaricus blazei and Agaricus bisporus fruit body powder was tested in vitro by stimulation peripheral blood mononuclear cells (PBMC's) and measuring interferon-gamma (IFN-γ) production by enzyme linked immunosorbent assay (ELISA). Titers of IFN-γ measured after incubation of activated cells with native A. blazei and A. bisporus glucans confirmed their characteristic immunostimulating activity on PBMC's. For A. bisporus the resulting β-(1,3)-glucan fragments of high molecular weight (MW>80 kDa) and the small fragments (MW lt 80 kDa) of A. blazei β-(1,3)-glucan remaining after enzymatic modification showed a significant enhancement of immunostimulating activity compared to the native glucans. The results confirmed that β-(1,3) bonds are the primary determinants of immunomodulatory activities and stimulation of IFN-γ synthesis. It indicated that primary structure is more important than the tertiary structure of the triple helix for their immunostimulating activity and synthesis of IFN-γ. The results confirmed that glucans of lower molecular weight are effective stimulatiors of IFN-γ synthesis., Hemijske modifikacije biološki aktivnih β-D-glukana viših gljiva su često neophodne kako bi se poboljšala njihova imunomodulatorska svojstva, a samim tim i njihova klinička primena. Cilj rada je ispitivanje imunomodulatorske aktivnosti nativnih miko-glukana, kao i glukana manjih molekulskih masa (MM lt 80 KD), nastalih nakon enzimske modifikacije 1,6 β-glukanazama i utvrđivanje koja je strukturna karakteristika primarno značajna za stimulaciju ćelija humanog imunog sistema. Imunomodulatorska aktivnost nativnih i enzimski modifikovanih glukana izolovanih iz praha plodonosnih tela gljiva Agaricus blazei i Agaricus bisporus testirana je in vitro stimulacijom mononuklearnih ćelija krvi iz perifernog krvotoka (PBMCPeripheral Blood Mononuclear Cell) molekulima glukana i merenjem količine sintetisanog interferona-gama (IFN-γ) od strane stimulisanih ćelija enzimoimunotestom (ELISA-enzyme linked immunosorbent assay). Merenjem titra IFN-γ nastalog nakon inkubacije aktiviranih ćelija sa nativnim A. blazei i A. bisporus glukanima potvrđeno je da su ovi molekuli ispoljili izraženo imunostimulativno dejstvo na PBMC ćelije. A. bisporus β-(1,3)-glukani velikih molekulskih masa (MM>80 kDa), kao i mali fragmenti A. blazei β-(1,3)-glukana (MM lt 80 kDa), nastali nakon enzimske modifikacije, ispoljili su značajno povećanje imunostimulativne aktivnosti u odnosu na nativne molekule. Ovim je potvrđeno da je za imunostimulativnu aktivnost i stimulaciju sinteze IFN-γ primarno značajno prisustvo β-(1,3)-glikozidnih veza, odnosno da je bitna primarna struktura, a ne konformacija trostrukog heliksa nativnih molekula. Dobijenim rezultatima je pokazano da su glukani manjih molekulskih masa efikasni stimulatori sinteze IFN-γ.", publisher = "Društvo za ishranu Srbije, Beograd", journal = "Hrana i ishrana", title = "Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms, Aktivni sastojci gljiva - ispitivanje promene imunomodulatorske aktivnosti enzimski modifikovanih glukana gljiva iz klase Basidiomycetes", pages = "20-15", number = "1-2", volume = "51", url = "https://hdl.handle.net/21.15107/rcub_agrospace_2185" }
Kozarski, M., Klaus, A.,& Nikšić, M.. (2010). Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms. in Hrana i ishrana Društvo za ishranu Srbije, Beograd., 51(1-2), 15-20. https://hdl.handle.net/21.15107/rcub_agrospace_2185
Kozarski M, Klaus A, Nikšić M. Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms. in Hrana i ishrana. 2010;51(1-2):15-20. https://hdl.handle.net/21.15107/rcub_agrospace_2185 .
Kozarski, Maja, Klaus, Anita, Nikšić, Miomir, "Active components of mushrooms: Changes in immnunomodulating activity of enzimatically modified glucans from Basidiomycetes mushrooms" in Hrana i ishrana, 51, no. 1-2 (2010):15-20, https://hdl.handle.net/21.15107/rcub_agrospace_2185 .