• Korean Journal of Pharmacognosy
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• v.43 no.1
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• pp.32-38
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• 2012

To examine the potentiation of Macrolepiota procera extracts (MPE-4) to act as adjuvant enhancing the tumor specific anti-tumor immune response, tumor vaccine prepared by boiling (HK vaccine) admixed with MPE-4 and immunized in mice. Vaccination of mice with HK vaccine in combination with MPE-4 resulted in higher inhibition in tumor metastasis compared with the mice of HK vaccine alone treatment against live syngeneic tumor cell challenge. The splenocytes from mice immunized HK vaccine mixed with MPE-4 was able to elicit a stronger cytotoxic T lymphocyte (CTL) response as compared with HK vaccine alone. In addition, the splenocytes from MPE-4 admixed HK vaccine immunized mice secreted a higher concentration of Th1 type cytokine such as IFN-${\gamma}$, and GM-CSF. Furthermore, the adoptive transfer of splenocytes from mice immunized HK vaccine and MPE-4 led to a more robust anti-tumour response than the HK vaccine alone. Overall, these results indicate that MPE-4 is a good candidate adjuvant of anti-tumor immune response.

• Korean Journal of Pharmacognosy
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• v.41 no.4
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• pp.275-281
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• 2010

To evaluate the immunomodulatory activity of a water extract (KMF-WE) of Korean mistletoe (Viscum album var. coloratum) fruit, we examined its ability to induce humoral and cellular immune response against keyhole limpet hemocyanine (KLH). Immunized mice with KLH admixed with KMF-WE (KLH/KMF-WE) showed significant induction of KLH-specific antibodies compared to mice immunized with KLH alone. The assay for determining isotypes of antibodies revealed that KMFWE augmented KLH-specific-IgG1 and -IgG2a production. In vitro T lymphocyte proliferation analysis against KLH revealed that the splenocytes of mice immunized with KLH/KMF-WE showed a significantly higher proliferative ability than those from mice immunized with KLH alone. The culture supernatants of splenocytes, which were harvested from mice immunized with KLH/KMF-WE, showed higher levels of both Th-1 type (IL-2, IFN-${\gamma}$) and Th-2 type (IL-4) cytokines in response to KLH stimulation compared to those from mice immunized with KLH alone. Also, in delayed-type hypersensitivity (DTH) assay, mice immunized with KLH/KMF-WE showed a significantly higher reaction to KLH than mice treated with KLH alone. These results suggest that KMF-WE possess immunoadjuvant activity to enhance both antigen-specific humoral and cellular immune responses against protein antigens (KLH).

• The Journal of Korean Medicine
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• v.32 no.3
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• pp.10-17
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• 2011

Objective: To evaluate the immune-stimulatory potential of extracts of Broussonetia kazinoki Siebold (BK) on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Material and Methods: C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. BK (100, 300 or 1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Further, the effects of BK on expression of cytokine mRNA in OVA-immunized mice splenocytes were evaluated by RT-PCR analysis. Results: BK significantly enhanced OVA-, LPS-, and Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). BK also significantly enhanced total IgM and OVA-specific IgG1 levels in plasma compared with the OVA control group. Moreover, BK up-regulated significantly the expression of mRNA level of IL-2 and IFN-${\gamma}$ in splenocytes. Conclusions: BK has immune-stimulating activity in an OVA-immunized mouse model system, enhancing the Th1 immune response. BK showed no cytotoxicity in this system, suggesting that BK may be a safe and effective adjuvant in humans.

• YAKHAK HOEJI
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• v.43 no.1
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• pp.48-54
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• 1999

Effects of zinc chloride (Zn) on the immune responses of lipopolysaccharide (LPS) were studied by using ICR mice. Mice were divided into 4 groups (10 mice/group), and Zn was given to the mice with i.p. injection at 0.3 mg/kg 5 times a week for 14 days, and 1 hr after Zn administration, LPS was given with i.p. injection at 5 mg/kg twice a week. Mice were immunized and challenged with sheep red blood cells (SRBC). Immunobiological responses were evaluated by humoral, cellular and nonspecific immunity. LPS treatment significantly increased the relative weights of spleen and thymus, hemagglutination titer (HA) and proliferation of splenocytes compared with those in controls, but significantly decreased the body weight gain. Zn treatment significantly increased proliferation of splenocytes and circulating leukocytes compared with those in controls. Combination of Zn and LPS significantly decreased the body weight gain and proliferation of splenocytes compared with those in controls. Combination of Zn and LPS significantly decreased HA and proliferation of splenocytes than in LPS alone. These findings indicate that zinc lowered the humoral immune responses of LPS.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.187-194
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• 2011

Objective : Astragali Radix (AR) and Cinnamomi Cortex (CC) are used to enhance immune response in Asian traditional medicine. Immuno-potentiation of the combination of AR and CC were evaluated on the cellular and humoral immune response using murine macrophage cell line (RAW 264.7) and OVA-immunized mice. Methods : This study was designed to investigate the immuno-potentiative effects of AR, CC, and AR with CC on nitric oxide synthesis in RAW 264.7 cells and proliferation and production levels of Intereukin-2 (IL-2) in mouse splenocytes. In addition, we evaluated the plasma-specific antibody responses and splenocyte proliferation on ovalbumin (OVA)-immunized mice treated with herbal extracts. Results : Combination treatment with AR and CC increased nitric oxide synthesis in RAW 264.7 cells and IL-2 level in splenocytes (p<0.001). Combination of AR and CC significantly enhanced the Concanavalin A- (Con A ; T cell mitogen) and lipopolysaccharide-(LPS ; B cell mitogen) induced splenocyte proliferation on the OVA-immunized mice. Combination of AR and CC also significantly enhanced plasma levels of OVA-specific IgG (p<0.01), IgG1 (p<0.05) and total IgM (p<0.01) compared with the OVA-immunized control group. Conclusion : These results suggest that combination of AR and CC could be used as therapeutic profile on activation of immune response.

• Journal of Sasang Constitution and Immune Medicine
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• v.22 no.3
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• pp.141-151
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• 2010

1. Objectives: Three herbal formulas (Yuldahanso-tang, Chungsimyonja-tang, and Taeeumjowi-tang) for Taeeumin were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. 2. Methods: This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results: Yuldahanso-tang and Chungsimyonja-tang increased NO synthesis in RAW 264.7 cells. Three herbal formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<.001). Three herbal formulas for Taeeumin also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. 4. Conclusion: These results suggested that three herbal formulas for Taeeumin could be used as stimulator of immune response.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.295-301
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• 2003

Background: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. In this study, we used a replication-deficient adenovirus containing CEA to study CTL induction in vitro after adenovirus-mediated gene transfer into DC. Methods: DC were obtained from mouse bone marrow and cultured with IL-4 and GM-CSF. For measuring CTL activity, splenocytes were harvested from the mice, which were immunized with DC that had been infected AdV-CEA or pulsed with CEA peptide. Untreated DC was used as a control. Splenocytes were re-stimulated in vitro with DC pulsed with CEA peptide for 7 days and CTL activity with CEA peptide-pulsed EL-4 cells were assessed in a standard $^{51}Cr$-release assay. The frequencies of antigen-specific cytokine-secreting T cell were determined with $mIFN-{\gamma}$ELISPOT. Results: DC infected with recombinant adenovirus expressing CEA induced CEA-specific CTL responses in vivo. Splenocyte induced from mice immunized with AdV-CEA-infected DC increase in the number of $IFN-{\gamma}$ secreting T cells compared with those from mice immunized with CEA peptide-pulsed DC. Conclusion: These results suggested that DC infected with recombinant adenovirus has advantages over other forms of vaccination and could provide an alternative approach vaccination therapies.

• The Journal of Korean Medicine
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• v.31 no.5
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• pp.112-123
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• 2010

Objectives: Three yin-tonifying formulae (Ssanghwa-tang, Yukmijihwang-tang and Jaeumganghwa-tang) were applied to investigate their immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in the murine macrophage cell line (RAW 264.7) and in ovalbumin (OVA)-immunized mice. Methods: This study was carried out in nitricoxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes in association with three herbal formulas. C57BL/6 mice were immunized intraperitoneally with OVA/aluminum ($100\;{\mu}g/200\;{\mu}g$/mouse) on days 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. Results: All three yin-tonifying formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Also, these herbal formulas all significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. Conclusion: These results suggested that the three yin-tonifying formulae could be used as stimulators of immune response.

• Journal of Sasang Constitution and Immune Medicine
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• v.22 no.4
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• pp.77-84
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• 2010

1. Objectives Yanggyuksanhwa-tang for Soyangin was applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. 2. Methods This study were carried out in nitric oxide (NO) synthesis on RAW 264.7 cells and cellular proliferation on mouse splenocytes. C57BL/6 mice were immunized intraperitonially with OVA/Alum (100 ${\mu}g$/200 ${\mu}g$) on day 1, 8, and 15. Yanggyuksanhwa-tang was administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results Yanggyuksanhwa-tang significantly enhanced cellular proliferation by LPS and Con A on splenocytes from OVA/Alum-immunized mice (p<.001). Yanggyuksanhwa-tang also significantly enhanced plasma OVA-specific IgG (p<.001), IgG1 (p<.001), and total IgM (p<.01) levels compared with the OVA/Alum group. 4. Conclusions These results suggested that Yanggyuksanhwa-tang for Soyangin could be used as immunopotent.

• YAKHAK HOEJI
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• v.47 no.3
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• pp.167-175
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• 2003

Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.