• 약학회지
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• 제47권3호
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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.

• IMMUNE NETWORK
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• 제11권6호
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• pp.383-389
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• 2011

Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

• IMMUNE NETWORK
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• 제23권6호
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• pp.46.1-46.16
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• 2023

Autoimmune hepatitis (AIH) affects all age group and occurs mainly in women. Pyroptosis is a novel programmed cell death featured with cell bursting and release of proinflammatory cytokines. A deeper understanding of AIH pathogenesis will contribute to novel therapy for AIH patients. Here, we aimed to investigate the role of IL-17 in immune-mediated liver injury. The levels of cytokines were measured by ELISA, and mRNA levels of STAT3 and IFN gamma-inducible protein 16 (IFI16) were detected by PCR. Expressions of STAT3, IFI16, gasdermin D and cleaved caspase-1 were measured by western-blotting. Immunohistochemical staining and transmission electron microscopy were applied to evaluate liver histopathological changes of the treated mice. Our results showed that the levels of IFI16 was increased in hepatocytes treated with IL-17 protein, and further elevated after STAT3-overexpressed (STAT3-OE) lentivirus treatment. The levels of IFI16 were reduced in hepatocytes treated with IL-17 neutralizing Ab (nAb), but were significantly increased after STAT3-OE treatment. Pyroptosis was observed in hepatocytes treated with IL-17 protein, and further cell damage was observed after STAT3-OE lentivirus treatment. Liver damage was alleviated in mice treated with IL-17 nAb, however sever damage was experienced after STAT3-OE lentivirus treatment. A binding interaction between IFI16 and STAT3 was detected in IL-17 treated hepatocytes. Glutathione transaminase activity was enhanced in concanavalin A-induced AIH mice compared to the control group (p<0.01). IL-17 plays an important role in activating STAT3 and up-regulating IFI16, which may promote the pyroptosis in AIH-related liver injury through STAT3-IFI16 axis.

• IMMUNE NETWORK
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• 제11권3호
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• pp.182-189
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• 2011

Background: Cytotoxic T lymphocytes (CTLs) appear to play an important role in the control and prevention of human cytomegalovirus (HCMV) infection. The pp65 antigen is a structural protein, which has been defined as a potential target for effective immunity against HCMV infection. Incorporation of an 11 amino acid region of the HIV TAT protein transduction domain (Tat) into protein facilitates rapid, efficient entry into cells. Methods: To establish a strategy for the generation of HCMV-specific CTLs in vitro, recombinant truncated N- and C-terminal pp65 protein (pp65 N&C) and N- and C-terminal pp65 protein fused with Tat (Tat/pp65 N&C) was produced in E.coli system. Peripheral blood mononuclear cells were stimulated with dendritic cells (DCs) pulsed with pp65 N&C or Tat/pp65 N&C protein and immune responses induced was examined using IFN-${\gamma}$ ELISPOT assay, cytotoxicity assay and tetramer staining. Results: DCs pulsed with Tat/pp65N&C protein could induce higher T-cell responses in vitro compared with pp65N&C. Moreover, the DCs pulsed with Tat/pp65 N&C could stimulate both of $CD8^+$ and $CD4^+$ T-cell responses. The T cells induced by DCs pulsed with Tat/pp65 N&C showed higher cytotoxicity than that of pp65-pulsed DCs against autologous lymphoblastoid B-cell line (LCL) expressing the HCMV-pp65 antigen. Conclusion: Our results suggest that DCs pulsed with Tat/pp65 N&C protein effectively induced pp65-specific CTL in vitro. Tat fusion recombinant protein may be useful for the development of adoptive T-cell immunotherapy and DC-based vaccines.

• Toxicological Research
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• 제32권2호
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• pp.149-158
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• 2016

Atopic dermatitis (AD) is a chronic inflammatory skin disease with a complex etiology that encompasses immunologic responses. AD is frequently associated with elevated immunoglobulin (Ig) E levels, and common environmental factors contribute to its pathogenesis. Several recent studies have documented the role of specific lactic acid bacteria in the treatment and prevention of AD in humans and mice. In this study, the efficacy of Duolac ATP, a probiotic preparation, was determined in a mouse model with AD-like skin lesions. Alterations in the cytokine levels and histological staining suggested the alleviation of AD. The in vivo test showed that T helper (Th)2 cytokines, IgE, interleukin (IL)-4, and IL-5, were significantly downregulated, whereas Th1 cytokines, IL-12p40 and interferon (IFN)-${\gamma}$, were upregulated in all groups of mice treated with Duolac ATP compared to that observed in the group of mice treated with 1-chloro-2,4-dinitrobenzene (DNCB) alone. Moreover, the scratch score decreased in all mice treated with Duolac ATP. Staining of the dorsal area of the mice in each group with hematoxylin and eosin and toluidine blue further confirmed the alleviation of AD in mice orally treated with Duolac ATP. These results suggest that Duolac ATP inhibits the development of AD-like skin lesions in NC/Nga mice by suppressing the Th2 cell response and increasing the Th1 cell response. Thus, Duolac ATP is beneficial and effective for the treatment of AD-like skin lesions.

• 혜화의학회지
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• 제18권2호
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• pp.63-79
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• 2009

Atopic dermatitis induced NC/Nga mice were used to investigate the efficacy of Hyunggaeyunkyotanggamibang(HYGB) on the recovery of dermatitic symptoms through its influence on the immune related factors. The results are as below: 1. HYGB treated group showed improvement of atopic dermatitis with naked eye observation, and significant decrease of dermatitis index was observed after 14 weeks. 2. HYGB treated group showed significant decrease of the ratio of CCR3+, B220+/IgE+, and CD11b+/Gr-1+ immune cells in dorsal skin by 41.7%, 21.5%, and 23.8%, respectively. 3. HYGB treated group showed an increase of CD19+ immune cells by 10.3% in PBMC, whereas CD3+, CD3+/CD69+, NKT+ immune cells were decreased by 4.3%, 42.9%, and 21.7%, respectively. 4. HYGB treated group showed an increase in the expression of IFN-$\gamma$ in the serum by 514.3%. However, the expressions of IL-4, IL-5, IL-6, IL-13, TNF-$\alpha$, MCP-1 and RANTES were decreased by 21.2%, 69.8%, 90.5%, 28.7%, 72.2%, 26.1%, and 19.9%, respectively. Also, the expression of IgE was decreased by 44.3%. 5. HYGB treated group showed a decrease of the expression of IL-4 and IL-5 by 43% and 44.3%. The results above indicated that HYGB clinically used for atopic dermatitis treatment has objective validity, and therefore can be provided as the basic data for EBM(Evidence based medicine) construction for anti-allergic and anti-inflammatory studies.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.309-314
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• 2013

Aim: Brain tumors almost universally have fatal outcomes; new therapeutics are desperately needed and will only come from improved understandins of glioma biology. Methods: Exosomes are endosomally derived 30~100 nm membranous vesicles released from many cell types. Examples from GL26 cells were here purified using density gradient ultracentrifugation and monitored for effects on GL26 tumor growth in C57BL/6j mice (H-2b). Lactate dehydrogenase release assays were used to detect the cytotoxic activity of CD8+T and NK cells. Percentages of immune cells producing intracellular cytokines were analyzed by FACS. Results: In this study, exosomes from murine-derived GL26 cells significantly promoted in vivo tumor growth in GL26-bearing B6 mice. Then we further analyzed the effects of the GL26 cells-derived exosomes on immune cells including CD8+T, CD4+T and NK cells. Inhibition of CD8+T cell cytotoxic activity was demonstrated by CD8+T cell depletion assays in vivo and LDH release assays in vitro. The treatment of mice with exosomes also led to a reduction in the percentages of CD8+T cells in splenocytes as determined by FACS analysis. Key features of CD8+T cell activity were inhibited, including release of IFN-gamma and granzyme B. There were no effects of exosomes on CD4+T cells and NK cells. Conclusion: Based on our data, for the first time we demonstrated that exosomes from murine derived GL26 cells promote the tumor growth by inhibition of CD8+T cells in vivo and thus may be a potential therapeutic target.

• 대한한의학회지
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• 제32권3호
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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.

• 한국발생생물학회지:발생과생식
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• 제21권1호
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• pp.35-46
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• 2017

The process of spontaneous abortion involves a complex mechanism with various cytokines, growth factors, and hormones during the pregnancy. However, the mechanism underlying spontaneous abortion by pro- and anti-inflammatory cytokines in the serum during the pregnancy is not fully understood. Therefore, the purpose of this study was to examine the relationship between the serum levels of pro- and anti-inflammatory cytokines and spontaneous abortion using the $CBA/j{\times}DBA/2$ mouse model. Serum levels of pro-inflammatory cytokines, such as $IFN-{\gamma}$, $IL-1{\alpha}$ and $TNF-{\alpha}$ were not increased in abortion model mice, but anti-inflammatory cytokines, such as IL-4, IL-13 and IL-1ra were decreased compared to normal pregnant mice. In addition, serum levels of chemokine, such as SDF-1, G-CSF, M-CSF, IL-16, KC and MCP-1 were decreased in abortion model mice compared to normal pregnant mice. However, the expression levels of nesfatin-1/NUCB2 mRNA and protein in the uteri of implantation sites were significantly higher in abortion model mice than normal pregnant mice. These results suggest that uterine nesfatin-1/NUCB2 expression may be down-regulated by inflammatory cytokines and chemokines in the serum of pregnant mice. Moreover, this study suggests the possibility that nesfatin-1/NUCB2 expressed in the implantation sites may be associated with the maintenance of pregnancy.

• Parasites, Hosts and Diseases
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• 제55권5호
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• pp.505-512
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• 2017

Toxoplasma gondii cathepsin C proteases (TgCPC1, 2, and 3) are important for the growth and survival of T. gondii. In the present study, B-cell and T-cell epitopes of TgCPC1 were predicted using DNAstar and the Immune Epitope Database. A TgCPC1 DNA vaccine was constructed, and its ability to induce protective immune responses against toxoplasmosis in BALB/c mice was evaluated in the presence or absence of the adjuvant ${\alpha}-GalCer$. As results, TgCPC1 DNA vaccine with or without adjuvant ${\alpha}-GalCer$ showed higher levels of IgG and IgG2a in the serum, as well as IL-2 and $IFN-{\gamma}$ in the spleen compared to controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). Upon challenge infection with tachyzoites of T. gondii (RH), $pCPC1/{\alpha}-GalCer$ immunized mice showed the longest survival among all the groups. Mice vaccinated with DNA vaccine without adjuvant (pCPC1) showed better protective immunity compared to other controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). These results indicate that a DNA vaccine encoding TgCPC1 is a potential vaccine candidate against toxoplasmosis.