Cholesterol as an inbuilt immunoadjuvant for a lipopeptide vaccine against group A Streptococcus infection.

Peptide-based vaccines can trigger highly specific immune responses, although peptides alone are usually unable to confer strong humoral or cellular immunity. Consequently, peptide antigens are administered with immunostimulatory adjuvants, but only a few are safe and effective for human use. To overcome this obstacle, herein a peptide antigen was lipidated to effectively anchor it to liposomes and emulsion. A peptide antigen B cell epitope from Group A Streptococcus M protein was conjugated to a universal T helper epitope, the pan DR-biding epitope (PADRE), alongside a lipidic moiety cholesterol. Compared to a free peptide antigen, the lipidated version (LP1) adopted a helical conformation and self-assembled into small nanoparticles. Surprisingly, LP1 alone induced the same or higher antibody titers than liposomes or emulsion-based formulations. In addition, antibodies produced by mice immunized with LP1 were more opsonic than those induced by administering the antigen with incomplete Freund's adjuvant. No side effects were observed in the immunized mice and no excessive inflammatory immune responses were detected. Overall, this study demonstrated how simple conjugation of cholesterol to a peptide antigen can produce a safe and efficacious vaccine against Group A Streptococcus - the leading cause of superficial infections and the bacteria responsible for deadly post-infection autoimmune disorders.

Effects of different processing methods on the antioxidant and immune stimulating abilities of garlic.

In this study, we determined the antioxidant and immune stimulating abilities of a garlic product developed by freeze drying, heat drying, and solid-state fermentation of heat-dried garlic. Lactobacillus plantarum KCTC21004 and Leuconostoc mesenteroides KCTC13302 were used for the sample fermentation. The optimum conditions for fermentation were 50% (v/w) moisture, a fermentation time of 48 hr and a temperature of 37°C. Heat-dried garlic samples fermented with L. plantarum KCTC21004 (HD21004) and L. mesenteroides KCTC13302 (HD13302) showed the highest flavonoid contents while heat-dried garlic (HD) had the lowest flavonoid content. HD21004 contained the highest phenolic compounds, showed the highest antioxidant activity and demonstrated a strong immune stimulating effect while freeze-dried garlic showed the lowest flavonoid and polyphenolic contents. Overall, the heat-dried garlic samples (fermented and unfermented) contained about three times more S-Allylcysteine (SAC) than the freeze-dried samples (FD). The current study demonstrates that heat drying and subsequent fermentation of garlic with L. plantarum KCTC21004 can improve its therapeutic effects.

The Production and Correlation of Silica Induced Proinflammatory Cytokines and TGF-beta from Monocytes of Balb/C Mice / 결핵

BACKGROUND: Chronic inhalation of silica induces the lung fiborsis. The alveolar macrophages ingest the inhaled silica they liberate the pro-inflammatory cytokines such as IL-i/A IL-C, TNF-a and fibrogenic cytokines, TGF-beta and PDGF. Cytokines liberated from macrophage have pivotal role in pulmonary fibrosis. There is a complex cytokine network toward fibrosis. However, the exact roles and the interaction among the proinflammatory cytokines and TGP-beta, a fibrogenic cytokine, have not been defined, yet In this study, we investigated silica induced IL-1/beta, IL-6, TNF-alpha and TGF-beta production and the effect of IL-1/beta, IL-6, TNF-alpha on the production of TGF-beta from lung macrophages of Balb/C mice. METHOD: We extracted the lung of Balb/C mice and purified monocytes by Percoll gradient method. Macrphages were stimulated by silica (SiO2) in the various concentration for 2, 4, 8, 12, and 24 bows. The supernatants were used for the measurement of protein levels by bioassay, and cells for the levels of mRNA by in situ hybridization. RESULTS: The production of IL-6 was not observed till 4 hours, and reached the peak levels at S horns after stimulation of silica. The production of TNF-alpha increased from 2 hours and reached the peak levels at 4 hours after stimulation of silica. The spontaneous TGF-beta production reached the peak levels at 24 hours. TNF-alpha upregulated the silica induced TGF-beta production Silica induced TGF-beta production was hooked by pretreated anti-TNF-alpha antibody. In situ hybridization revealed the increased positive signals at 4 hours in IL-6, at 4 hours in TNF-alpha and 12 hours in TGP-beta. CONCLUISON: The results above suggest that silica induced the sequential production of IL-6, TNF-alpha and TGF-beta from macrophages and TNF-alpha upregultaes the production of TGF-beta from silica-induced macrophages.