E per CCR5 Synonyms experiment, total three independent experiments). (C) Gene expression in spleens
E per experiment, total 3 independent experiments). (C) Gene expression in spleens was measured 24 hrs after fucoidan injection. Information will be the average of analyses of six independent samples (2 mice per experiment, total three independent experiments). (D) Fucoidan was injected to C57BL6 mice in conjunction using a neutralizing anti-IL-1223p40 antibody or control rat IgG as well as the identical injections have been repeated three day later. Intracellular IFN-c expressions in CD4 or CD8 T cells from these mice have been analyzed on a flow cytometer. (E) Serum IFN-c levels from mice described in (D). All data are representative of six samples from three independent experiments. doi:10.1371journal.pone.0099396.gPLOS One particular | plosone.orgFucoidan Functions as an Adjuvant In VivoFigure four. Fucoidan gives an adjuvant effect on OVA-induced B and T cell responses. C57BL6 mice were immunized i.p with PBS, OVA or OVA fucoidan on days 0, 15, 30. On day 35, serum OVA-specific IgG1 (A) and IgG2a (B) concentrations were measured by ELISA. P,0.05, P, 0.01 versus OVA group. #P,0.05, ##P,0.01 versus PBS group. (C) Splenocytes were harvested from immunized mice on day 35, and re-stimulated with or with out OVA (50 mgml) for 4 days. Cell proliferation from re-stimulated splenocytes was measured. (D) IFN-c concentrations inside the above splenocytes culture supernatants were shown. (E) CD44 expression on CD4 or CD8 T cells was analyzed on a flow cytometer (left panel). Percentage of CD44 cells in CD4 or CD8 T cells was shown (appropriate panel). All information are representative of six samples from 3 independent experiments. doi:10.1371journal.pone.0099396.gfound that fucoidan did not affect the generation of IFN-cproducing CD4 or CD8 T cells (information not shown). We also showed that CD4 and CD8 T cell activation by fucoidan is dependent on IL-12 and that fucoidan can stimulate DCs to make IL-12.Importantly, fucoidan induced up-regulation of IL-12p40 mRNA on the other hand it did not affect IL-23p19 mRNA levels. Moreover, serum levels of IL-23 had been not up-regulated by fucoidan administration but IL-12p70 was up-regulated, suggesting thatPLOS One | plosone.orgFucoidan Functions as an Adjuvant In VivoFigure 5. Fucoidan promotes antigen presentation and antigen-specific T cell proliferation in vivo. (A) C57BL6 mice had been injected with PBS, OVA or OVA fucoidan for 24 hrs, along with the expression levels of MHC class I and II on the gated Lineage2CD11c cDCs in splenocytes from thesePLOS 1 | plosone.orgFucoidan Functions as an Adjuvant In CXCR6 custom synthesis Vivomice were analyzed. (B) Purified CD8 T cells from OT-I or CD4 T cells from OT-II mice have been labeled with CFSE and transferred into CD45.1 congenic mice, and 24 hrs later, mice were injected with PBS, OVA or OVA fucoidan. Immediately after three day treatment, splenocytes from these mice have been stained for CD45.2 to recognize the donor OT-I or OT-II cells and the proliferation of those cells was determined by CFSE dilution. All data are from analyses of 6 individual mice every group (2 mice per experiment, total three independent experiments). doi:10.1371journal.pone.0099396.gthe advertising impact of fucoidan on Th1 and CTL responses may possibly be accomplished by enhancing IL-12 production from DCs. Fucoidan stimulates macrophage and DC activation via scavenger receptor-A (SR-A) in in vitro studies [19,23,32], and it can be most likely that fucoidan may well stimulate in vivo spleen cDCs by engaging SR-A. Activation of SR-A results in human peripheral blood DC (PBDC) maturation that subsequently promotes Th1 responses [23]. DCs are known to prime.
