| Summary: | 碩士 === 臺灣大學 === 園藝學研究所 === 98 === Nano-scaled functional ingredients with different sizes, shapes and material properties have many applications in biological transport and hence the bioavailability. In spite of what has been achieved so far, cellular response between nano-scaled functional ingredient and target cells remains mostly unknown. Immunomodulatory protein named Ling Zhi-8 (LZ-8) extracted from the mycelia of Ganoderma lucidum has been cloned and expressed at Saccharomyces cerevisiae expression system as a recombinant protein rLZ-8. In the present study, suspensions of gold nanoparticles (GNPs) with stable surface rLZ-8 coating were prepared using electrostatic adsorption strategy. As size distribution from 10 to 100 nm, the quantity of rLZ-8 binding on GNPs was obviously correlated with particle size, which significantly enhanced the multivalency of rLZ-8 being conjugated on GNP surface.
Treatment of mouse peritoneal macrophages with GNP-LZ-8 significantly increased IL-1β cytokine production and surface CD80/86, MHC class II molecule expression as compare with the univalent free form LZ-8. Although the induction of cytokine and surface molecule activities was enhanced for all GNP-LZ-8, particle with larger size exhibited greater difference on both cytokine production and surface costimulatroy molecule expression on macrophages. Furthermore, side scatter (SSC) variation of macrophages after GNP-LZ-8 uptake revealing a granularity enhancement as well as an increase of GNP-LZ-8 size. GNP-LZ-8 treatment further indicated the enhancement of cell proliferation, IL-2 and INF-γ cytokine production as compare to univalent free form rLZ-8 on splenoctes. Moreover, free form rLZ-8 could activate the surface expression of CD25 on CD3+ splenocytes, while GNP-LZ-8 treatment elicited more significant upregulation on both CD3+ and CD3- cells. Among splenocyte treated with various sizes GNP-LZ-8, the greatest difference was observed in 30 and 50 nm GNP-LZ-8-treated cells. Taken together, these data implied the induction of diverse size-dependent immunomodulatory responses between phagocyte and non-phagocyte.
Base on these results, nano-size modification should no longer be consider only as a passive rule of delivery or bio-transportation, but could also play an active role in mediating biological effects. The findings presented here may assist in the design and development of nano-scaled functional ingredient.
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