• YAKHAK HOEJI
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• v.45 no.6
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• pp.617-622
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• 2001

A protein-polysaccharide fraction of a Korean wild mushroom Psathyrella velutina, PVP, was prepared and its antitumor immunomodulatory activity was investigated. When PVP was administered once daily for seven days from day 1 to day 7 into male ICR mice implanted with 1 $\times$ 10$^{5}$ cells of sarcom 180 tumor cells into the peritoneum on day 4, it inhibited the growth of sarcoma 180 cells by 92.8%. In XTT assay, PVP also exhorted in vitro anti-proliferation activity on U-937, a human monoblastoid cell line, as well as sarcoma 180 cells. PVP showed marked stimulatory activity on the immune system in that it induced the accumulation of PEC (the stimulation index, Sl=4.90 at 100 mg/kg), stimulated the BALB/c mouse splenic lymphocytes to form lymphoblasts (Sl=5.75 at 100$\mu\textrm{g}$/ml), and upergulated the expression of CD25 molecules. All these results strongly support that PVP exhorts its antitumor activity through stimulation of the immune system as well as anti-proliferative activity on the tumor cells.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• IMMUNE NETWORK
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• v.11 no.1
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• pp.50-58
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• 2011

Background: Epstein-Barr virus associated gastric lymphoepithelioma-like carcinoma (LELC) is characterized by the intensive infiltration of lymphoid cells, the presence of EBV, and the better prognosis over typical adenocarcinoma. Thus, it was assumable that viral latent proteins may be responsible for the recruitment of a certain T cell repertoire to EBV-associated gastric carcinoma. Methods: To examine above possibility, EBV gene expression in gastric carcinoma tissues and usage of TCR among the tumor infiltrating lymphocytes were analyzed. Results: EBV specific DNA and EBERs RNA were detected in 4 out of 30 patients. RT-PCR analysis revealed that all 4 of EBV-positive tumor tissues expressed EBNA1 mRNA and BARTs and LMP2a was detected only one sample out of 4. However, the EBNA2 and LMP-1 transcripts were not detected in these tissues. $CD8^+$ T cells were the predominant population of infiltrating lymphocytes in the EBV-positive gastric carcinoma. According to spectra type analysis of infiltrating T cells, 10 predominant bands were detected by TCR $V{\beta}$ CDR3 specific RT-PCR from 4 EBV-positive tumor tissues. Sequence analysis of these bands revealed oligoclonal expansion of T cells. Conclusion: These findings suggest that clonally expanded T cells in vivo might be a population of cytotoxic T cells reactive to EBV-associated gastric carcinoma.

• Journal of Veterinary Science
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• v.25 no.1
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• pp.4.1-4.14
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• 2024

Background: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. Objectives: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. Methods: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. Results: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4⁺ and CD 8⁺ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 10^6.9 times the median tissue culture infectious dose of LI or 2 × 10⁹ colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. Conclusions: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.484-488
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• 1994

Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

• Korean Journal of Food Science and Technology
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• v.40 no.5
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• pp.574-579
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• 2008

Propolis is the generic term for the resinous substance collected by honey bees from a variety of plant sources. In this study, we have assessed the immunomodulatory properties of propolis (P) and fermented-propolis (FP) in BALB/c mice. Mice were subjected to gavage once a day (for 14 days) with 50, 100, 200 mg/kg body weight P, FP, or vehicle. Lymphocytes were isolated from the spleen and mesenteric lymph nodes (MLN) and the immune cell proportions, proliferative activities, and cytokine production were evaluated. The P- and FP-administration induced similar, but differential, alterations in the percentage of immune cell populations and their biological functions, including cytokine production and NK cell cytotoxicity. The proportion of$ CD4^+$ and $CD8^+$ T cells in the spleen was increased slightly in the P- and FP-administered mice as compared to the vehicle-treated mice. In MLN, the percentage of $CD4^+$ T cells was increased significantly in the 200 mg/kg P-treated mice. The mice which were treated with P and FP evidenced significantly increased interferon-$\gamma$ and interleukin-4 production in concanavalin A-stimulated splenocytes, whereas the production of theses cytokines was not shown to be induced by P-treatment. In addition, NK cell activity was also increased dramatically by the administration of P and FP. Collectively, these findings showed that P and FP are wide-spectrum immunomodulators, which may modulate both innate and adaptive immune responses.

• Journal of Korean Neurosurgical Society
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• v.38 no.2
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• pp.126-131
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• 2005

Objective : The choice of tumor antigen for dendritic cell[DC]-loading has still been an unresolved problem in the DC-based vaccine strategies against malignant gliomas that has not been found well-characterized tumor specific antigens. In this study, we compare tumor-specific T cell response induced by glioma apoptotic body[GAB]-pulsed DCs to response induced by glioma cell lysate-pulsed ones quantitatively. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor and interleukin[IL]-4 from peripheral blood mononuclear cells[PBMCs] of HLA-A2 positive healthy donors were cultured. Each GABs and glioma cell lysate generated from HLA-A2 positive T98G glioblastoma cells were co-incubated with DCs. $CD8^+$ T lymphocytes isolated from PBMCs of same donors were cultured in media containing IL-2 and either stimulated by GAB- or lysate-pulsed DCs three times at a weekly interval. The interferon[IFN]-${\gamma}$ concentrations of each cell culture supernate were measured by enzyme immunoassay technique. Cytolytic activity of the generated cytotoxic $CD8^+$ T cells either stimulated with GAB- or lysate-pulsed DCs was determined by a standard 4-h $^{51}Cr$-release assay. Results : IFN-${\gamma}$ production and cytolytic activity of effector T cells stimulated by GAB-pulsed DCs were significantly higher than those of T cells stimulated by lysate-pulsed ones. Conclusion : These results indicate the choice of antigen is a critical determinant in the induction of antitumor immunity against malignant glioma. Antigen preparations from GABs represent a promising alternative to glioma cell lysate in DC-based glioma vaccine strategies.

• Korean Journal of Medicinal Crop Science
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• v.26 no.1
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• pp.72-81
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• 2018

Background: Polysaccharides are the most important functional constituent in Astragalus membranaceus. The purpose of the present study was to evaluate the effect of polysaccharides isolated from the aboveground parts of A. membranaceus (AMA) and polysaccharides isolated from the roots of A. membranaceus (AMR) immune function by modulated cytotoxic T cell and Th1- and Th2-related cytokines kinetics. Methods and Results: Sprague-Dawley rats were randomly divided into exhaustive exercise case groups and non-exercise case, AMA and AMR samples were administered orally for 30 days (500 mg/kg/day and 10 mg/kg/day, respectively) and were compared to those rats in the groups fed commercial sports drink (SPD) and vehicle. Both exhaustive exercise groups and non-exercise groups had a lower ratio of $CD4^+$ and $CD8^+$ cells in the spleens of the rat fed AMA and AMR compared to those in the rats fed SPD and vehicle group. These results suggested that AMA and AMR promote an increase in the proportion of cytotoxic T cells. The IL-4-producing T lymphocytes decreased significantly in the AMR (10 mg/kg/day) group compared to SPD and vehicle, whereas the AMA group increased the IL-4 concentration more than the SPD and vehicle in exhaustive exercise group. However, the populations of IFN-${\gamma}$-producing T lymphocytes of AMR and AMA increased. AMA decreased the concentration of IFN-${\gamma}$ to inhibit the Th1 response and thereby increased the concentration of IL-4 to induce a Th2 response that was related to humoral immunity in the non-exercise group. Conclusions: These results showed that, in addition to Th1/Th2 regulation, AMR and AMA played an important immuno-modulatory role after exhaustive exercise-induced Th1/Th2 lymphocyte imbalance, which might be correlated with cytokine producing immunoregulatory cells.

• Reproductive and Developmental Biology
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• v.31 no.1
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• pp.1-6
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• 2007

The nonobese diabetic / severe combined immune deficiency (NOD/SCID) has been used for determination of proliferation and differentiation of hematopoietic stem cells as xenotransplantation animal model. In this study, we transplanted porcine hematopoietic cells from bone marrow into NOD/SCID mice via intravenous injection to confirm the activity of differentiation and proliferation for porcine hematopoietic cells in vivo. Interestingly, we observed the result of high efficiency with pig T lymphocytes in hematopoietic organs, liver, spleen lymph node, and bone marrow in NOD/SCID mice. The porcine $CD3^{+}$ T cells were detected with $5.4{\pm}1.9%$ in bone marrow, $15.4{\pm}7.3%$ in spleen, $21.3{\pm}1.4%$ in liver, and $33.5{\pm}32.8%$ in lymph node of NOD/SCID mice at 6 weeks after trans-plantation Furthermore, immunohistochemical analysis showed the high engraftment of porcine T lymphocytes in spleen of NOD/SCID mice. Our data suggest that NOD/SCID mice are excellent animal model to determinate the generation md function of pig T lymphocytes.

• IMMUNE NETWORK
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• v.5 no.3
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• pp.172-178
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• 2005

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. However, it is known that the induction of immune response to CEA is very difficult because CEA is a self-antigen expressed in fetal cells and weakly expressed in normal colorectal epithelial cells. To enhance anti-tumor immunity specific for CEA, recombinant CEA protein was modified using listeriolysin O (LLO) for endosomal lysis and trans activator of transcription (Tat) domain for transducing extracellular proteins into cytoplasm. Methods: After immunization using dendritic cells pulsed with Tat-CEA, both Tat-CEA and LLO, and both Tat-CEA and Tat-LLO, antibody titer to CEA and LLO, cytotoxic T lymphocyte activity and the frequency of IFN-${\gamma}$ producing T lymphocytes were measured. Results: Immunization using DC pulsed with both Tat-CEA and Tat-LLO protein showed the increasement of production of CEA-specific antibody in serum, cytotoxic T lymphocyte activity, the frequency of IFN-${\gamma}$ secreting T cells, compared with DC pulsed with both Tat-CEA and LLO. Furthermore the ratio of CD8+T cell to $CD4^+$ cell among CEA-specific T cells was increased in group pulsed with both Tat-CEA and Tat-LLO. Conclusion: These results suggested that DC vaccine using Tat-LLO could be used for the development of effective immunotherapy for the treatment of tumor.