• 약학회지
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• 제49권6호
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• pp.484-489
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• 2005

Streptococcus pmeumoniae is the leading cause of pneumonia and bacterial meningitis. The current polysaccharide vaccine has been reported ineffective in elderly adults and children less than 2 years of age. Thus, in recent many researchers have been focused on a different approach, DNA vaccine. In our laboratory we developed a Streptococcus pneumoniae DNA (SPDNA) vaccine. This SPDNA vaccine was formulated by inserting the region encoding part of the capsule in the S. pneumoniae into the LAMP-1. In present work, with use of the SPDNA vaccine we attempted to establish a certain methodology useful for evaluation of effectiveness and immunoresponse of a DNA vaccine. Results showed that the subcutaneous route was the most effective for production of antisera specific for S. pneumoniae in mice. By isotyping analyses, IgM, IgGl, IgG2a, and IgG2b were determined. In addition, INF-$\gamma$ and IL-4 were predominantly detected. Combination of those data resulted in a pattern of IgGl < IgG2a=IgG2b and INF$\gamma\>$ >IL-4, which indicates the inmmunity towards the Thl response predominantly; furthermore, the SPDNA vaccination induced resistance of the CD4+T lymphocyte-depleted mice against disseminated pneumococcal infection. These data appear to be possibly due to activation of CDS8+T cell-activation. Taken together, this methodology can be applied for evaluating efficacy and mode of action of a DNA vaccine as minimum critera.

• IMMUNE NETWORK
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• 제2권1호
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• pp.41-48
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• 2002

Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제28권1호
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• pp.135-142
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• 2015

This study was conducted to evaluate the effect of inclusion of propolis extraction residue in the feed of broilers from 1 to 21 d of age on phagocytic activity of macrophages, cutaneous basophil hypersensitivity response to phytohemagglutinin, antibody production against Newcastle disease, lymphoid organ weight and hematological profile and to determine the optimal level of inclusion. 120 chicks, reared in metabolism cages until 21 days of age, were distributed in a completely randomized design, with five treatments (0%, 1%, 2%, 3%, and 4% of propolis residue) and six replications. The relative weight of thymus and monocyte percentage were affected by propolis residue, with a quadratic response (p<0.05) and lowest values estimated at 2.38% and 2.49%, respectively. Changes in relative weight of cloacal bursa and spleen, percentage of lymphocyte, heterophil, basophil, eosinophil, and heterophil:lymphocyte ratio, antibody production against Newcastle disease, phagocytic activity of macrophages and the average number of phagocytosed erythrocytes were not observed. The nitric oxide production with regard to positive control (macrophages+erythrocytes) decreased linearly (p<0.05) with increased doses of propolis residue. The remaining variables of nitric oxide production (negative control - macrophages, and difference between the controls) were not affected by propolis residue. The cutaneous basophil hypersensitivity response to phytohemagglutinin as determined by the increase in interdigital skin thickness exhibited a quadratic response (p<0.05), which predicted a lower reaction response at a dose of 2.60% of propolis residue and highest reaction response after 43.05 hours of phytohemagglutinin injection. The inclusion of 1% to 4% of propolis extraction residue in broiler diets from 1 to 21 days of age was not able to improve the immune parameters, despite the modest changes in the relative weight in thymus, blood monocyte percentage, nitric oxide concentration, and interdigital reaction to phytohemagglutinin.

• 한국식품영양과학회지
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• 제44권11호
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• pp.1759-1762
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• 2015

시판 막걸리에서 분리한 균주(Lactobacillus paracasei ssp. paracasei M13-65-3)로 타락을 제조하였다. M13-65-3균주를 이용하여 제조한 타락 추출물에 의하여 T lymphocyte인 Jurkat 세포의 증식이 촉진되었고 피부세포의 멜라닌 생성이 억제되었다. 본 연구의 결과는 타락이 면역 증강 기능성과 피부색소 조절 기능성을 보유할 가능성을 제시하고 있다.

• IMMUNE NETWORK
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• 제2권2호
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• pp.109-114
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• 2002

Background: To investigate the role of sympathetic nervous system (SNS) in moxibustion-induced immunomodulation, the effects of chemical sympathectomy on moxibustion-induced changes in splenic NK cell cytotoxicity, T and B cell proliferation were studied in Sprague-Dawley male rats. Methods: Chemical sympathectomy was achieved with intraperitoneal injection of 6-hydroxydopamine 50 mg/kg/day for 3 successive days. Direct moxibustion (6-minute interval, 9 moxa ball, each of which weighing 0.007 g and burning for 40 seconds) was applied on unilateral anterior tibial muscle region where Zusanli (ST36) acupoint is located, once a day for 7 successive days. NK cell cytotoxicity was measured by $4hr-^{51}Cr$ release assay. Mitogen-induced lymphocyte proliferation was analyzed by [$^3H$]-thymidine incorporation assay. Results: NK cell cytotoxicity was suppressed by moxibustion, more in sympathectomized rats than in vehicle-treated rats. T cell proliferation induced by concanavalin A was not affected by moxibustion. B cell proliferation induced by lipopolysaccharide showed no significant change in vehicle-treated rats, but an increase in sympathectomized rats by moxibustion. Sympathectomy alone induced augmentation of NK cell cytotoxicity and suppression of T cell proliferation. Conclusion: These results suggest that SNS has no direct relation with moxibution-induced immunomodulation but has an important role in the mechanism to keep the homeostasis of immune system by tonically inhibiting excessive changes of various immune components.

• 대한의생명과학회지
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• 제24권1호
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• pp.1-8
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• 2018

${\mu}MT$ knockout mice are genetically deficient in the transmembrane domain of mu chain of the immunoglobulin M (IgM) heavy chain, resulting in the absence of mature B cells. ${\mu}MT$ knockout mice is an in vivo model system used to clarify the role of B cells in various diseases. Enteropathogenic Escherichia coli (EPEC) induces acute and chronic diarrheal disease, especially in children of developing countries. The formation of attaching and effacing (A/E) lesion is a prominent pathogenic factor in the intestinal epithelium of EPEC infection. The A/E lesion is modulated by genes located on the pathogenic island locus of enterocyte effacement (LEE) which encode a type III secretion system (T3SS) and A/E lesion-related effector proteins. Citrobacter rodentium is a murine pathogen utilized in studying the pathogenic mechanisms of EPEC in human infections. Citrobacter rodentium produce A/E lesion to attach to intestinal epithelium, thus providing a murine model pathogen to study EPEC. Several studies have investigated the pathogenesis of Citrobacter rodentium in the ${\mu}MT$ knockout mice. In this review, we introduce the ${\mu}MT$ murine model in the context of C. rodentium pathogenesis and describe in detail the role of B cells and antibodies in this disease.

• Journal of Acupuncture Research
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• 제18권1호
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• pp.186-201
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• 2001

Purpose ; In order to study the effect of Houttuyniae herbal acupuncture on growth of melanoma B 16 and immunity in mice, the Control group with normal saline acupunchure after subcutaneous inoculation of B16BL6 cells, the Sample I with Houttuyniae herbal acupuncture manufactured by distilled water extraction method after subcutaneous inoculation of B16BL6 cells and the Sample II with Houttuyniae herbal acupuncture manufactured by alcohol extraction method after subcutaneous inoculation of B16BL6 cells were divided. Methods ; To evauluate the effect of Houttuynias herbal acupuncture on growth of melanoma B16 in mice, the weight of mouse melanoma, body weight, spleen index( $\sqrt(Weight of spleen/Body weight){\times}100$), lymphocytes numbers in mouse peripheral blood and spleen tissue, the percentage of CD4+ T cells, CD8+ and the ratio of CD4+/CD8+ in mouse peripheral blood and spleen tissue are measured. Results ; In study of the weight of mouse melanoma, Houttuyniae herbal acupuncture groups(Sample I, Sample II) showed statistically significant inhibitory effect. And in study of effect of reduction of change in body weight of mouse, Sample I showed statistically significant inhibitory effect, too. In study of reduction of spleen index, lymphocytes numbers in mouse spleen tissue, and the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse peripheral blood, Houttuyniae herbal acupuncture groups(Sample I, Sample II ) showed with statistically significant inhibitory effect. Also, Sample I showed inhibitory effect of reduction of the percentage of CD8+ T cells in mouse peripheral blood and spleen tissue and Sample II showed inhibitory effect of reduction of the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse spleen tissue with statistical significance. Conclusions ; The inhibitory effect of reduction of lymphocytes numbers in mouse peripheral blood and IL-2 productivity, Houttuyniae herbal acupuncture groups(Sample I, Sample II) didn't show significant effect.

• IMMUNE NETWORK
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• 제21권6호
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• pp.44.1-44.15
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• 2021

Tumor peptides associated with MHC class I molecules or their synthetic variants have attracted great attention for their potential use as vaccines to induce tumor-specific CTLs. However, the outcome of clinical trials of peptide-based tumor vaccines has been disappointing. There are various reasons for this lack of success, such as difficulties in delivering the peptides specifically to professional Ag-presenting cells, short peptide half-life in vivo, and limited peptide immunogenicity. We report here a novel peptide vaccination strategy that efficiently induces peptide-specific CTLs. Nanoparticles (NPs) were fabricated from a biodegradable polymer, poly(D,L-lactic-co-glycolic acid), attached to H-2K^b molecules, and then the natural peptide epitopes associated with the H-2K^b molecules were exchanged with a model tumor peptide, SIINFEKL (OVA_257-268). These NPs were efficiently phagocytosed by immature dendritic cells (DCs), inducing DC maturation and activation. In addition, the DCs that phagocytosed SIINFEKL-pulsed NPs potently activated SIINFEKL-H2K^b complex-specific CD8⁺ T cells via cross-presentation of SIINFEKL. In vivo studies showed that intravenous administration of SIINFEKL-pulsed NPs effectively generated SIINFEKL-specific CD8⁺ T cells in both normal and tumor-bearing mice. Furthermore, intravenous administration of SIINFEKL-pulsed NPs into EG7.OVA tumor-bearing mice almost completely inhibited the tumor growth. These results demonstrate that vaccination with polymeric NPs coated with tumor peptide-MHC-I complexes is a novel strategy for efficient induction of tumor-specific CTLs.

• Genomics & Informatics
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• 제20권1호
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• pp.11.1-11.20
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• 2022

Vibrio harveyi belongs to the Vibrio genus that causes vibriosis in marine and aquatic fish species through double-stranded DNA virus replication. In humans, around 12 Vibrio species can cause gastroenteritis (gastrointestinal illness). A large amount of virus particles can be found in the cytoplasm of infected cells, which may cause death. Despite these devastating complications, there is still no cure or vaccine for the virus. As a result, we used an immunoinformatics approach to develop a multi-epitope vaccine against most pathogenic hemolysin gene of V. harveyi. The immunodominant T- and B-cell epitopes were identified using the hemolysin protein. We developed a vaccine employing three possible epitopes: cytotoxic T-lymphocytes, helper T-lymphocytes, and linear B-lymphocyte epitopes, after thorough testing. The vaccine was developed to be antigenic, immunogenic, and non-allergenic, as well as having a better solubility. Molecular dynamics simulation revealed significant structural stiffness and binding stability. In addition, the immunological simulation generated by computer revealed that the vaccination might elicit immune reactions in the actual life after injection. Finally, using Escherichia coli K12 as a model, codon optimization yielded ideal GC content and a higher codon adaptation index value, which was then included in the cloning vector pET2+ (a). Altogether, our experiment implies that the proposed peptide vaccine might be a good option for vibriosis prophylaxis.