• 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.

• Journal of Microbiology and Biotechnology
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• 제28권1호
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• pp.122-135
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• 2018

Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of $CD8^+$ lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

• 대한한방부인과학회지
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• 제19권4호
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• pp.1-16
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• 2006

Purpose : The purpose of this research was to investigate the effects of Kamibojoongikkitang (KBT) on the immune cells in C57BL/6 mice. Methods : KBT (500mg/kg) was administerd p.o. once a day for 7 days. Results : KBT decreased the cell viability of thymocytes in vivo and in vitro system and decreased the cell viability of splenocytes in vivo, but increased the viability of splenocytes in vitro system. In addition, KBT did not affect the population of helper T (Th) cells and cytotoxic T (Tc) cells in thymocytes and decreased the population of T- and B-lymphocytes and the population of Th and Tc cells in splenocytes. Furthermore, KBT did not affect the production of ${\gamma}%-interferon and interleukin-4 in splenocytes. KBT increased the production of nitric oxide in vivo but decreased the production of nitric oxide in vitro system. KBT enhanced the phagocytic activity of peritoneal macrophages in vivo, but decreased the phagocytic activity in vitro. Conclusion : KBT has an inhibitory action on the specific immune response via decrease of the cell viability of thymocytes and splenocytes and has a potent action on the non-specific immunity via increase of phagocytic activity of peritoneal macrophages.

• Journal of Periodontal and Implant Science
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• 제24권2호
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• pp.357-375
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• 1994

The purpose of this study was to investigate the differences of histochemical characteristics in inflammatory fibrous gingival hyperplasia (FGH), phenytoin-induced gingival hyperplasia(PIGH), idiopathic gingival hyperplasia(IDGH) and control groups (healthy and inflammatory gingiva) by immunohistochemical method with various antibodies and histomorphological analysis. In immunohistochemical finding, antibodies to inflammatory cells (T/B lymphocytes, macrophages, other monocytes), proliferating cell nuclear antigen(PCNA), epidermal growth factor(EGF), factor VIII, and type I collagen were used. 1. The inflammatory infiltrates in FGH were less than those in inflammatory gingiva. The composition of inflammatory cells of PIGH was similar with that of FGH. IDGH showed a similar histologic findings with healthy gingival tissue. 2. In FGH, the number of fibroblasts and newly-formed collagen fibers was increased. No significant increase of fibroblasts and the dense accumulation of thick collagen fibers were seen in PIGH. The increase of fibroblasts and the dense accumulation of thick collagen were seen in IDGH. 3. PCNA-positive cells were localized mainly in the area accumulated with inflammatory cells and blood vessels, significantly increased in all hyperplastic tissue groups, and distributed evenly in IDGH. 4. The distribution of EGF were not observed in healthy gingiva but detected locally in area with confluent blood vessels,without significant difference between the other tissue groups. This results suggest that inflammation plays a significant role in inducing hyperplastic change of gingival tissue. While in DIGH, drug itself as well as inflammation seems to attribute to hyperplastic change.

• Asian-Australasian Journal of Animal Sciences
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• 제26권11호
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• pp.1577-1582
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• 2013

The effects of dietary supplementation of selenium (Se), iodine (I), and a combination of both on the blood haematology, serum free thyroxine (FT4) and free triiodothyronine (FT3) hormones and glutathione peroxidase enzyme (GSH-Px) activity were examined on twenty four (7 to 8 months old, $22{\pm}1.17$ kg live weight) Kacang crossbred male goats. Animals were randomly assigned to four dietary treatments (6 animals in each group). Throughout 100 d of feeding trial, the animals of control group (CON) received a basal diet, while the other three groups were offered basal diet supplemented with 0.6 mg/kg diet DM Se (SS), or 0.6 mg/kg diet DM I (PI), or a combination of both Se and I, each at 0.6 mg/kg diet DM (SSPI). The haematological attributes which are haemoglobin (Hb), red blood cell (RBC), packed cell volume (PCV), mean cell volume (MCV), white blood cells (WBC), band neutrophils (B Neut), segmented neutrophils (S Neut), lymphocytes (Lymph), monocytes (Mono), eosinophils (Eosin) and basophils (Baso) were similar among the four treatment groups, while serum levels of Se and I increased significantly (p<0.05) in the supplemented groups. The combined dietary supplementation of Se and I (SSPI) significantly increased serum FT3 in the supplemented animals. Serum GSH-Px activity increased significantly in the animals of SS and SSPI groups. It is concluded that the dietary supplementation of inorganic Se and I at a level of 0.6 mg/kg DM increased serum Se and I concentration, FT3 hormone and GSH-Px activity of Kacang crossbred male goats.

• 한국연초학회지
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• 제13권1호
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• pp.19-26
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• 1991

Ames test using special strains which are histidine requiring mutant of Salmonella typhimufium , is widely utilized as short-term bioassay to evaluate the genotoxicity of chemicals. This method requires the liver microsome(5-9 fraction) to provide mammalian metabolism of the compounds, Therefore, the mutagenic potency of the chemicals is affected by not only the intrinsic properties of them but also the efficiency of the in vitro microsomal activation system. For this reason, the complex mixtures such as environmental pollutants from occupational sources, natural products or cigarette smoke condensates(CSC) , might be often appeared the false results. Induction of sister chromatid exchange(SCE) in cultured cells is known as another sensitive and powerful tool for the measurement of genotoxicity and the method has also an advantage which is able to apply to the in vitro and in vivo systems. In the present study, the inducibilities of revertant colonies in tester strain TA98 and SCE in human Iymphocytes by positive controls and total particulated materials(TPM) obtained from various brand(domastic and imported) cigarettes were compared in order to investigate whether the results in Ames test are in agreement with those in SCE analysis. The mutagenic activities of well known mutagens such as B(a)P showed excellent dose-response in the both methods although the induction mechanism was different each other, but cyclophosphamide resulted such effect only in SCE analysis. Most TPM tested showed a similar pattern in the mutagenic activities in those methods. However, only two(one imported brand and one domestic sample cigarettes) among the TMP obtained from various cigarettes appeared the higher induction in SCE than Ames test.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.398-405
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• 2002

Bifidobacteria have been previously shown to stimulate the immune functions and cytokine production in macrophages and T-lymphocytes. Accordingly, the RAW 264.7 murine macrophage cell line was used to assess the effects of Bifidobacterium on the proliferation and cytoskeleton reorganization of the cells. Cytokine production after exposure to Bifidobacterium was also monitored in both whole cells and cell-free extracts. When RAW 264.7 cells were cultured for 24 h in the presence of heat-killed Bifidobacterium bifidum BGN4, the proliferation of macrophages was slowed down in a dose-dependent manner and cell differentiation was observed by staining with the actin-specific fluorescent dye, rhodamin-conjugated phalloidin. Although EL-4 cells, a T-cell line, stimulated RAW 264.7 cells to produce TNF-${\alpha}$ and IL-6, the stimulatory activity of B. bifidum BGN4 decreased as the EL-4 cell number increased. When disrupted and fractionated BGN4 was used, the whole cell fraction was more effective than the other fractions for the TNF-${\alpha}$ production. In contrast, the cell-free extract exhibited the highest IL-6 production level among the fractions, which was evident even at a $1{\mu}g/ml$ concentration. The current results demonstrate that Bifidobacterium induced differentiation of the macrophages from the fast proliferative stage and that the cytokine production was differentially induced by the whole cells and cell-free extracts. The in vitro approaches employed herein are expected to be useful in further characterization of the effects of bifidobacteria with regards to gastrointestinal and systemic immunity.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1431-1438
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• 2008

Immunomodulating activities of water-soluble exopolysaccharides (LL-EX) obtained from submerged mycelial culture of Lentinus lepideus were studied and their effectiveness was compared with lipopolysaccharide (LPS). The influence of the LL-EX on macrophage cellular lysosomal enzyme activity was to stimulate up to 267%, 392%, and 464% at the level of 10, 50, and $100{\mu}g/ml$, respectively. When the LL-EX was further fractionated into LL-Fr.I and Fr.II by Sepharose CL-6B gel chromatography, the cellular lysosomal enzyme activity of LL-Fr.II (2.1-fold) was higher than Fr.I (1.2-fold). Moreover, both LL-Fr.I and Fr.II stimulated the cytokines IL-1$\beta$, TNF-$\alpha$, and IL-6 in macrophages. In mixed lymphocyte reaction, LL-Fr.I and Fr.II enhanced the splenocyte proliferation up to 1.2-fold and 1.4-fold ($50{\mu}g/ml$), respectively, stimulating only T lymphocytes. The fractions of LL-EX not show any direct toxicity against human gastric adenocarcinoma cell (AGS). The molecular masses of LL-Fr.I and Fr.II were estimated to be about 1,986 kDa and 21 kDa, respectively. The total sugar and protein contents of the two fractions were 84.97% and 69.88%, and 15.03% and 30.12%, respectively. The sugar and amino acid compositions of the LL-Fr.I and Fr.II were also analyzed in detail.

• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.169-173
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• 2009

Ginsan, a Panax ginseng polysaccharide that contains glucopyranoside and fructofuranoside, has immunomodulatory effects. Although several biologic studies of ginsan have been performed, its effects on dendritic cells (DCs), which are antigen-presenting cells of the immune system, have not been studied. We investigated the immunomodulatory effects of ginsan on DCs. Ginsan had little effect on DC viability, even when used at high concentrations. Ginsan markedly increased the levels of production by DCs of IL-12 and TNF-${\alpha}$, as measured by ELISA. To examine the maturation-inducing activity of ginsan, we measured the surface expression levels of the maturation markers MHC class II and CD86 (B7.2) on DCs. It is interesting that ginsan profoundly enhanced the expression of CD86 on DC surfaces, whereas it increased that of MHC class II only marginally. In $^3H$-thymidine incorporation assays, ginsan-treated DCs stimulated significantly higher proliferation of allogeneic $CD4^+$ T lymphocytes than did medium-treated DCs. Taken together, our data demonstrate that ginsan stimulates DCs by inducing maturation. Because DCs are critical antigen-presenting cells in immune responses, this study provides valuable information on the activities of ginsan.

• 한국임상수의학회지
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• 제16권2호
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• pp.454-462
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• 1999

Corticosteroids have long been used for anti-inflammatory, anti-rheumatoid and other purposes in hospital. These effects may be due to inhibit immune reaction. So the animal given corticosteroids was more susceptible to infection because of immunosuppressive effect of corticosteroids. The purpose of this study was to investigate the effects of prednisolone on the lymphocyte subset in the spleen, immunoglobulin in serum, spleen weight, thymus weight and total WBC in peripheral blood. Mice were randomized into 3 groups. Each group has 24 mice. The small dosage group were given by 4 mg/kg/day of prednisolone for 4 days and the large dosage group were given by 8 mg/kg/day respectively. Prednisolone was suspended in saline and orally administered. Mice in control group were given saline alone. Eight mice in each group were sacrificed every week after administration of predisolone. The weight of thymus and spleen were mesured immediately. Lymphocytes were taken from spleen and these cells were analysed by flow cytometry. Also the concentration of total immunoglobulins in serum were assayed by enzyme-linked immunosorbant assay (ELISA). T cell, T-helper cell and T-cytotoxic cell were all significantly (P<0.05) decreased at 1 week after administration of predisolone and at 2 weeks they recovered similarly to that of control. Population of B cell showed various distribution. The concentration of total immunoglobulins in serum was not changed significantly. The weight ratio of spleen to body decreased significantly (P＜0.05) during predisolone administration but increased at 1 week later, Eventually the weight ratio was recovered to that of control at 2 weeks. The weight ratio of thymus to body decreased significantly (P<0.05) by prednisolone and recovered gradually up to normal ratio 2 weeks later.