• IMMUNE NETWORK
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• v.15 no.2
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• pp.83-90
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• 2015

Evaluation and screening of vaccines against tuberculosis depends on development of proper cost effective disease models along with identification of different immune markers that can be used as surrogate endpoints of protection in preclinical and clinical studies. The objective of the present study was therefore evaluation of subcutaneous model of M.tuberculosis infection along with investigation of different immune biomarkers of tuberculosis infection in BALB/c mice. Groups of mice were infected subcutaneously with two different doses : high ($2{\times}10^6CFU$) and low doses ($2{\times}10^2CFU$) of M.tuberculosis and immune markers including humoral and cellular markers were evaluated 30 days post M.tuberculosis infections. Based on results, we found that high dose of subcutaneous infection produced chronic disease with significant (p<0.001) production of immune markers of infection like $IFN{\gamma}$, heat shock antigens (65, 71) and antibody titres against panel of M.tuberculosis antigens (ESAT-6, CFP-10, Ag85B, 45kDa, GroES, Hsp-16) all of which correlated with high bacterial burden in lungs and spleen. To conclude high dose of subcutaneous infection produces chronic TB infection in mice and can be used as convenient alternative to aerosol models in resource limited settings. Moreover assessment of immune markers namely mycobacterial antigens and antibodies can provide us valuable insights on modulation of immune response post infection. However further investigations along with optimization of study protocols are needed to justify the outcome of present study and establish such markers as surrogate endpoints of vaccine protection in preclinical and clinical studies in future.

• IMMUNE NETWORK
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• v.15 no.2
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• pp.73-82
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• 2015

Respiratory syncytial virus (RSV) infection is recognized by the innate immune system through Toll like receptors (TLRs) and retinoic acid inducible gene I. These pathways lead to the activation of type I interferons and resistance to infection. In contrast to TLRs, very few studies have examined the role of NOD-like receptors in viral recognition and induction of adaptive immune responses to RSV. Caspase-1 plays an essential role in the immune response via the maturation of the proinflammatory cytokines IL-$1{\beta}$ and IL-18. However, the role of caspase-1 in RSV infection in vivo is unknown. We demonstrate that RSV infection induces IL-$1{\beta}$ secretion and that caspase-1 deficiency in bone marrow derived dendritic cells leads to defective IL-$1{\beta}$ production, while normal RSV viral clearance and T cell responses are observed in caspase-1 deficient mice following respiratory infection with RSV. The frequencies of IFN-${\gamma}$ producing or RSV specific T cells in lungs from caspase-1 deficient mice are not impaired. In addition, we demonstrate that caspase-1 deficient neonatal or young mice also exhibit normal immune responses. Furthermore, we find that IL-1R deficient mice infected with RSV exhibit normal Th1 and cytotoxic T lymphocytes (CTL) immune responses. Collectively, these results demonstrate that in contrast to TLR pathways, caspase-1 might not play a central role in the induction of Th1 and CTL immune responses to RSV.

• IMMUNE NETWORK
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• v.11 no.3
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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.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.1
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• pp.23-30
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• 2013

The purpose of this study was to evaluate the effects of several types of bibimbab (a Korean traditional meal of mixed rice with assorted vegetables), on various immune activities. Compared to control animals in a mouse model (given hamburgers), the oral administration of a portion of bibimbab containing wild plants significantly increased splenic B/T, thymic Th lymphocyte subpopulations, serum IFN-${\gamma}$ production, and enhanced hemagglutination titers up to 300%. Also, a consumption of mushroom-bulgogi bibimbab and Jeonju-style bibimbab markedly decreased compound 48/80-induced systemic anaphylaxis (immediate hypersensitivity), while bibimbab with wild plants inhibited SRBC-induced delayed type hypersensitivity. These results suggest that bibimbab with wild plants both up-regulate on immune activities and have anti-allergenic properties.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2094-2103
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• 2017

Aging is associated with distinct changes in immune cells and a decline in immune function, leading to increased susceptibility to infection and reduced responses to vaccination. Certain strains of lactic acid bacteria exert beneficial effects on the immune system. Previously, we reported that Weissella cibaria JW15 isolated from kimchi possesses immune stimulatory activity in vitro. In the present study, we further investigated whether oral administration of JW15 improves immune function in aged mice. Eighteen-month-old female mice were administered JW15 daily at low (JW15-L; $1{\times}10^8CFU/mouse$) or high dosage (JW15-H; $1{\times}10^9CFU/mouse$), or with Lactobacillus rhamnosus GG (LGG) using oral gavage. Two-month-old female mice were included as healthy young mice. After 4 weeks, the mice were euthanized and immune profiles were analyzed using whole blood and spleen. In complete blood count analysis, the numbers of white and red blood cells were significantly increased in the JW15-L group compared with those in the old mouse (OM) control group. In addition, administration of either JW15 of LGG resulted in higher numbers of splenocytes in comparison with the OM group. Furthermore, proliferative potentials were higher in all probiotic groups than OM. Cytokines such as IFN-${\gamma}$ and IL-6 were secreted at higher levels in splenocytes isolated from JW15-fed mice than in OM control mice. Similarly, mRNA expression of various cytokines was altered in the JW15 groups. Collectively, these results suggest that JW15 supplementation induces immunomodulatory effects in aged mice and indicate JW15 as a potential probiotic strain to improve immune function in aged animals.

• Journal of Haehwa Medicine
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• v.23 no.2
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• pp.5-13
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• 2015

Objectives : The anti-inflammatory and antioxidant effects of water extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) were investigated. The effects of SSJ and GSJ were compared. Methods : We performed cell viability assay in HaCaT cells and RAW 264.7 cells using 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide(MTT) assay. We measured chemokines(regulated on activation normal T-cell expression and secreted ; RANTES/CCL5, interferon-inducible protein; IP-10/CXCL10, macrophage-derived chemokine; MDC/CCL22) in HaCat cells, also we measured cytokines (tumor necrosis factor-${\alpha}$; TNF-${\alpha}$, interleukin-6; IL-6) and nitric oxide(NO) production in RAW 264.7 cells using enzyme-linked immunosorbent assay(ELISA) and NO assay. Western blot assay was used to evaluate the expression for inducible nitric oxide synthase(iNOS) in RAW 264.7 cells. Results : SSJ and GSJ did not affect the cell viability at the concentrations treated ($0-800{\mu}g/ml$). As a result of SSJ and GSJ treatment in HaCat cells stimulated by TNF-${\alpha}$(10 ng/ml) and interferon(IFN)-${\gamma}$(10 ng/ml), the production of RANTES and IP-10 was inhibited significantly. However there was no significant difference in the secretion of MDC. And in RAW 264. 7 cells stimulated by lipopolysaccharide(LPS, $1{\mu}g/ml$), SSJ and GSJ treatment significantly inhibited the secretion of TNF-${\alpha}$ and IL-6 and the production of NO. The expression of iNOS was also decresed by SSJ and GSJ treatment in RAW 264. 7 cells. Compared with SSJ, GSJ was superior to SSJ in inhibition of RANTES, IP-10, TNF-${\alpha}$, IL-6 and NO production at the concentration of $200{\mu}g/ml$. Conclusion : Both SSJ and GSJ have anti-inflammatory and antioxidant effects. And GSJ has better effects than SSJ.

• IMMUNE NETWORK
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• v.4 no.4
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• pp.229-236
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• 2004

Background: Disialoganglioside GD2 is a tumor-associated antigen that is overexpressed on tumor cells of neuroectodermal origin, such as melanoma, small cell lung carcinoma and neuroblastoma. Immunity against GD2 has anti-tumor activities, but GD2 is poorly immunogenic. Anti-idiotypic antibodies that mimic GD2 may induce more effective immune responses than GD2 antigen itself, because they are protein antigens and are known to be able to break immune tolerance. In our previous study, we produced anti-idiotypic antibodies mimicking GD2 (3A4 and 3H9), which induced humoral immunity. However, cellular immunity is essential to eradicate tumor cells in vivo as well as humoral immunity. In the present study, we investigated whether these anti-idiotypic antibodies 3A4 and 3H9 could induce cellular immunes responses. Methods: BALB/C mice were immunized with anti-idiotypic antibody 3A4 or 3H9, or normal mouse IgG as a negative control. Lymphoproliferative responses, cytokine production responses, and delayed-type hypersensitivity reactions were measured in mice immunized with the anti-idiotypic antibodies. Results: Both the anti-idiotypic antibody 3A4 and 3H9 induced GD2-specific lymphoproliferative responses and $IFN-{\gamma}$ production of lymph node lymphocytes in BALB/C mice. Only anti-idiotypic antibody 3H9 induced significant GD2-specific delayed-type hypersensitivity in the mice. Conclusion: These results show that anti-idiotypic antibodies 3A4 and 3H9 have the potentiality of inducing GD2-specific cellular immune responses that cannot be induced by the native antigen GD2 itself.

• Korean Journal of Medicinal Crop Science
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• v.17 no.4
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• pp.273-279
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• 2009

GATA-binding protein-3 (GATA-3) and T-box expressed in T-cells (T-bet) are now considered as master transcription factors involving Th cell differentiation, but the roles of these factors are still uncertain in vivo. This study was conducted to investigate the expression of these transcription factors in the liver damage induced by carbon tetrachloride ($CCl_4$) in rats. In this study, liver damage were induced with Scutellaria baicalensis Georgi water extracts (SBW) and followed for 4 weeks. The expression of GATA-3 and T-bet protein in liver damage induced by $CCl_4$ and the serum levels of immunoglobulin A (IgA), IgE were studied after 4 weeks of treatment. We found that effect of SBW on IFN-$\gamma$, STAT1, pSTAT1 and T-bet was decreased in vivo. Several genes were demonstrated to be IL-4 inducible prior to the discovery of STAT6. $CCl_4$+SBW group was significantly lower than $CCl_4$ group in IL-4, STAT6, pSTAT6 and GATA-3. Our data indicate that cytokine protein production were increased in $CCl_4$ group and $CCl_4$+SBW group. From these results, water extracts obtained from Scutellaria baicalensis Georgi may have an immunoregulatory effect in the liver induced by $CCl_4$ of rats.

• IMMUNE NETWORK
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• v.5 no.1
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• pp.36-44
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• 2005

Background: The anti-tumor therapeutic effect of autologous tumor cell lysate pulseddendritic cells (DCs) was studied for non-immunogenic and immune suppressive lung cancer model. To test the possibility as an adjuvant therapy, minimal residual disease model was considered in mouse in vivo experiments. Methods: Syngeneic 3LL lung cancer cells were inoculated intravenously into the C57BL/6 mouse. Autologous tumor cell (3LL) or allogeneic leukemia cell (WEHI-3) lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, tumor formation in the lung and the tumor-specific systemic immunity were observed. Tumor-specific lymphocyte proliferation and the IFN-${\gamma}$ secretion were analyzed for the immune monitoring. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with tumor cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor formation was suppressed in 3LL tumor cell lysate pulsed-DC treated group, while 3LL-specific immune stimulation was minimum. WEHI-3-specific immune stimulation occurred in WEHI-3 lysate-pulsed DC treated group, which had no correlation with tumor regression. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs as an adjuvant therapy for minimal residual disease state of lung cancer. The significance of immune modulation in DC therapy including the possible involvement of NK cell as well as antigen-specific cytotoxic T cell activity induction was discussed.

• Journal of Pharmacopuncture
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• v.7 no.2
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• pp.43-56
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• 2004

Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu($BL_{21}$) and Chung- wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant differences. 2. For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3. For survival time, all of experiment groups showed 11.1% increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5. For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6. For expression of cytokine mRNA using RT-PCR, significant increase of IL.-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all off experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effccts in anti-cancer and immune improvement if increasing concentration.