• YAKHAK HOEJI
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• v.56 no.6
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• pp.366-371
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• 2012

In this present study, we determined whether or not there is an immunoadjuvant effect of ochnaflavone, a biflavone isolated from Lonicera japonica. As an antigenic source, the cell wall (CACW) of Candida albicans, a fungal pathogen, was used. CACW consists of 95% carbohydrate (mannan). In the experiments, BALB/c mice were immunized with emersion forms of CACW combined with or without ochnaflavone (Och) in the presence of IFA containing mineral oil or CACW alone. Then, the amounts of antisera collected from these mice groups were measured by the ELISA method. Data from these experiments showed that CACW combined with Och (CACW/Och/IFA) provoked the production of antisera app. 2.2 or 5 times more than the corresponding CACW/IFA or CACW alone (CACW/DPBS), respectively, in mice (P<0.05). We further examined the immune response type induced by Och. Analysis of the values of the IgG1/IgG2a ratios obtained from IgG isotyping revealed that Och induced Th2-immunity more dominantly than Th1. This finding was confirmed by cytokine profile. CACW/Och/IFA formulation induced IL-4 (Th2-type cytokine) more than IFN${\gamma}$ (Th1-type cytokine) as compared with CACW/IFA and CACW/DPBS formulations (P<0.05). All data combined, Och appears to have an immunoadjuvant activity that may convert Th1 immunity into Th2 immunity.

• YAKHAK HOEJI
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• v.59 no.4
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• pp.170-176
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• 2015

In vaccine development, the major points may be induction of effective and increased levels of antibody production. This is especially the case when the antigenic sources are carbohydrates. For many years, thus, we have researched various types of formulations such as liposomal and conjugate vaccines. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested if platycodin D (PLD) from Platycodon Radix have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that in the murine model of antibody production, CACW combined with PLD [CACW/PLD/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CACW/PLD/IFA], the antibody production was 1.4 times as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity maintaining the initial titers of antibody as compared to the CFA formula. Cytokine profiling with the antisera displayed that the PLD produced both Th1 and Th2 immunoresponses, but Th1 dominant was much greater (P<0.05). Furthermore, [CACW/PLD/IFA] formula enhanced resistance of mice against disseminated candidiasis, whereas the CFA had no such effect. In conclusion, PLD has an immunologic activity, which is protective against the disease. Thus, PLD can be a goof candidate for a new immunoadjuvant in development of the fungal vaccine.

• YAKHAK HOEJI
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• v.54 no.6
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• pp.494-499
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• 2010

We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.

• YAKHAK HOEJI
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• v.57 no.1
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• pp.1-7
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• 2013

Induction of effective and increased levels of antibody production may be major points in vaccine development. This is especially the case when the antigenic sources are carbohydrates. Thus, in our Lab various types of formulations such as liposomal and conjugate vaccines have been researched. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested whether ginsenosides Re (a panaxdiol) and Rg1 (a panaxtriol) from Panax ginseng have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that Rd and Rg1 caused LPS-treated B lymphocytes to be proliferative. Rd had greater proliferation activity than that of Rg1. In the murine model of antibody production, CACW combined with Rd [CACW/Rd/IFA] or Rg1 [CACW/Rg1/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CFA/Rd/IFA], the antibody production was almost twice as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity as compared to the CFA formula. In conclusion, Rd and Rg1 have an immunologic activity, and yet Rd can be a better candidate than Rg1 for a new immunoadjuvant.

• YAKHAK HOEJI
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• v.52 no.6
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• pp.494-499
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• 2008

The role of antibody in the fungal infections is controversial. However, our previous reports showed a certain epitope in Candida albicans cell wall (CACW) induces protective antibody. A major problem is that the epitope isolation requires tremendous time with high cost. This aspect led us to investigate a simple way inducing protective antibodies against C. albicans. In the present study, we determined if $18{\beta}$-glycyrrhetinic acid ($18{\beta}$-GA) from Glabrae Radix (a family of Leguminosae) has immunoadjuvant activity. Data displayed that the $18{\beta}$-GA suppressed proliferations of both T- and Blymphocytes at high concentrations, whereas below 20 ${\mu}M$ concentration the compound supported the proliferations. These observations indicate that $18{\beta}$-GA has immunoregulatory activity. Based on this observation, an immunoadjuvant effect was examined at the low concentration. Results from animal experiments showed that CACW combined with or without $18{\beta}$-GA produced the anti-C. albicans antiserum in mice. Nevertheless, the CACW combined with $18{\beta}$-GA formula only protected mice against disseminated candidiasis (P<0.05). These data implicate that $18{\beta}$-GA has immunoadjuvant activity, which may provoke the CACW antigen to induce protective antibody. Currently, we are investigating possible mechanism of how the $18{\beta}$-GA provokes such protective immunity against the disseminated disease.

• YAKHAK HOEJI
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• v.56 no.1
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• pp.20-25
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• 2012

In Northeast Asia, Paeoniae Radix has been used in treatments of inflammation-causing diseases such as arthritis for many centuries. Paeoniflorin, one of the principle bioactive monoterpene glucosides from the paeony root, is reported to be mostly responsible for the effectiveness of the treatments. However, the anti-inflammatory effect of a monoterpene, paeoniflorigenone (PFG) which partially has the moiety of paeoniflorin minus a glucose structure is unknown. Thus, the aim of this work was to investigate anti-inflammatory activity of PFG. For the investigation, PFG activity on the NO (nitric oxide) production from LPS-stimulated macrophages, and the anti-inflammatory effect was tested in the animal model of septic arthritis caused by Candida albicans, a major etiological agent for septic arthritis. For induction of the arthritis, mice were administered with an emulsion of C. albicans cell wall (CACW) mixed with Complete Freund's Adjuvant (CFA) via footpad-injection (Day 0); PFG at a dose of 0.5 or 1 mg/mouse (25 or 50 mg/kg of body-weight) was given to the animals on Day 3, 6, and 9; footpads were scored for arthritis. Moreover, the PFG effect on proliferation of T-lymphocyte that causes aggravation of arthritis was additionally tested. Data resulting from those tests showed that PFG inhibited the NO production from the stimulated macrophage in a dose dependent manner (P<0.05), indicating that PFG is an anti-inflammatory. To confirm the in-vitro results, anti-inflammatory activity of PFG was determined against C. albicans-caused septic arthritis. Data showed that PFG-treatment reduced footpad-swelling which indicates that PFG has anti-arthritic effect (P<0.05), which is therapeutic. The anti-arthritic effect appeared to be mediated by PFG suppression of T-cell proliferation. Ultimately, PFG, a monoterpene component, has anti-inflammatory activity analogous to paeoniflorin. The anti-inflammatory activity treats the septic arthritis due to a pathogenic fungus C. albicans.

• YAKHAK HOEJI
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• v.58 no.5
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• pp.343-348
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• 2014

In the present studies, we examined effect of chamomile flowers extract (CH-Ex), which has traditionally been used as antiphlogistics in Europe for many centuries, against Candida albicans-caused septic arthritis. Candida albicans is a major etiological agent among fungal septic arthritis. This effect was investigated in a murine model of the septic arthritis. That is, mice that were given an emulsion form of C. albicans cell wall (CACW) via footpad route were treated intraperitoneally with the CH-Ex for 3 times every 3 days. Degrees of the footpad-swellings were measured with dial gauger. Data showed that the CH-Ex resulted in the reduction of swelling. For instance, at Day 9 when swelling reached the highest peak, there was up to app. 60% reduction of edema in mice injected with the CH-Ex, compared to that of the control mice that received no treatment (P<0.05). This therapeutic anti-arthritic activity appeared to be mediated by inhibitions of NO (nitric oxide) production from activated RAW264.7 macrophages and proliferation of Con A-treated T lymphocytes. Analysis by HPLC revealed that the CH-Ex contained eight polyphenolic compounds including chlorogenic acid (CRA) and rutin. We have reported the CRA and rutin respectively have the anti-arthritic activity. This correlation implicates that CRA and rutin in the CH-Ex may be responsible for the activity. Combined all together, the CH-Ex has anti-arthritic activity against C. albicans-caused septic arthritis, possibly by inhibiting NO production and proliferation of T cells. This activity seems to be contributed by, at least, CRA and rutin among the compounds in the CH-Ex.