• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.552-559
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• 2003

The FimH subunit of type 1-fimbriated Escherichiu coli (E. coli) has been determined as a major cause for urinary tract infections. Thus, to produce a possible vaccine antigen against urinary tract infections, the fimIH gene was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. A fusion protein, based on fusing adhesin to the cholera toxin subunit A2B (CTXA2B), was induced with 0.01 mM isopropyl-${\beta}-D-thiogalactoside$ (IPTG) for 4 h at $37^{\circ}C$ to yield a soluble fusion protein. The fusion protein was then purified by affinity chromatography. The expressed fusion protein was confirmed by SDS-PAGE and Western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was also analyzed. The orderly-assembled fusion protein was confirmed by a modified $G_{Ml}-ganglioside$ ELISA, using antibodies to adhesin. The results indicated that the purified fusion protein was an adhesin/CTXA2B protein containing E. coli adhesin and the $G_{Ml}-ganglioside$ binding activity of CTXB. Accordingly, this adhesin/CTXA2B protein may be a potential antigen for oral immunization against uropathogenic E. coli.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.56-62
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• 2000

The hpa gene genetically linked to the ctxa2b gene was cloned into the pTED expression vector, and the constructed pTEDhpa/ctxa2b was transformed into Excherichia coli. The fusion protein, the adhesin fused to the cholera toxin subunit A2B (CTXA2B) subunit, was expressed to high levels as inclusion bodies in E. coli. The expressed protein was partially purified by washing the inclusion bodies with working solution containing 8M Urea and 0.1M DTT. Refolding of denatured fusion protein was carried out in the presence of glutathione redox buffer. The refolded fusion protein was purified by size exclusion chromatography. The expressed fusion protein was verified by SDS-PAGE, western blotting with antibodies to both antigenic components of adhesin and cholera toxin subunit B (CTXB), and its N-terminal amino acid sequence was analyzed. The orderly assembled fusion protein was confirmed by modified Gm1-ganglioside ELISA with Abs to adhesin. The results indicate that the purified fusion protein is an Adhesin/CTXA2B protein containing the H. pylori adhesin and $G_{m1}4-ganglioside binding activity of CTXB and the expressed fusion protein in E. coli could be easily purified by the refolding process, Its molecular weight was 168kDa as estimated by size exclusion chromatography. The Adhesin/CTXA2B protein may be used as a candidate antigen for oral immunization against H. pylori.

• 대한한의학방제학회지
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• 제13권1호
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• pp.145-159
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• 2005

Objective : Juglandis Semen has a function that to invigorate the lung and kidney. And It is commonly used as a supporting agent in the treatment of coughing and bronchitis. This study was performed to investigate the effect of oral administration of Juglandis Semen Extract (JSE) against the experimental asthma induced by ovalbumin. Methods : Asthma was induced to Balb/c mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the cell number in the BAL fluid. Concentrations of IL-4, IL-5 in splenoc yte were assessed by ELISA, IgG and IgE from serum were calculated by same method. Results : 1. Number of macrophage in BAL fluid was significantly decreased in JSE group compared with control group, but not eosinophil and lymphocyte. 2. Levels of IgG and IgE in serum were significantly decreased in JSE group compared with control group, respectively. 3. Concentration of IL-4 in culture supernatant of splenocyte was significantly decreased in JSE group compared with control group, but there was no significant in IL-5. Conclusion : We found that the effect of JSE extract in asthma was implicated in reduction of IL4released from Th2 cell, and decreases of IgG and IgE from plasma cell. These findings suggest that JSE can produce anti-asthmatic effect, which may play a role in allergen-induced asthma therapy.

• 대한한의학방제학회지
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• 제21권1호
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• pp.161-176
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• 2013

Objectives : To clarify the anti-arthritic activity of Ostericum koreanum Maxim. (OS) plus Aralia continentalis Kitagawa (AC) in vivo. Methods : All mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with OS plus AC once a day for 7 weeks. Oral administration of OS plus AC (200 or 50 mg/kg) significantly suppressed the progression of CIA, which extend is comparable to that of methotrexate (MTX, 0.3 mg/kg), a positive control. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. Results : Administration of OS plus AC significantly suppressed the progression of CIA and inhibited the production of TNF-${\alpha}$ and IL-6 in serum. The erosion of cartilage was dramatically reduced in mouse knees after treatment with OS plus AC. In conclusion, our results demonstrates that OS plus AC significantly suppressed the progression of CIA and that this action was characterized by the decreased production of IL-6, IFN-${\gamma}$ and collagen II specific antibody in serum, CD3+CD69+ T cells, MHC class II+/CD11c+ (in DLN), CD11b+Gr-1+ cells (in PBMC), CD11b +Gr-1+ cells, B220+/CD23+ (in paw joint). Conclusions : The the levels of IFN-${\gamma}$ in the culture supernatant of splenocytes stimulated with CD3/CD28 or collagen were dramatically decreased, while those of IL-4 was increased. In the serum of OS and AC-treated mice, the levels of IgM RA factor were decreased.

• 한국가금학회지
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• 제31권1호
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• pp.37-44
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• 2004

It has been recognized that the hen, like its mammalian counterparts, provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk, and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immune-incompetent newly hatched chick has, is through transmission of antibodies from the mother via the egg. Egg yolk, therefore, can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus, the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8∼20 mg of jmmunoglobulins (IgY) per ml or 136∼340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk, low cost antibodies at less than $10 per g compared to more than $20,000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine, public health, veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool, nutraceutical or functional food development, oral-supplementation for prophylaxis, and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed, the specific antibody binds, immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics, since today, more and more antibiotics are less effective in the treatment of infections, due to the emergence of drug-resistant bacteria.

• 동의생리병리학회지
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• 제19권5호
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• pp.1243-1250
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• 2005

This study was performed to investigate the effect of oral administration of deer horn against the asthma. Deer horn improves body metabolism and strengthens overall health, especially in elderly persons and young children. Additionally, it stimulates sexual function in females and can stimulate wound healing. Asthma was induced to Balblc mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the eosinophil number in the BALF. Concentrations of IL-4, IL-5 in BALF and splenocyte were assessed by ELISA, IgG and IgE from serum were calculated by same method. The number of eosinophil in BALF was not significantly changed in deer horn group compared with control group. Concentration of IL-4 in BALF was significantly decreased in deer horn group compared with control group. Levels of IL-5 from BALF and splenocyte were significantly decreased in deer horn group compared with control group, respectively. Concentrations of IgE and IgG in serum were significantly decreased in deer horn group compared with control group, separately. We found that the effect of deer horn extract in asthma was implicated in reductions of IL-4, IL-5 released from Th2 cell, and decreases of IgG, IgE from plasma cell. These findings suggest that deer horn extract can produce anti-asthmatic effect, which may play a role in allergen-induced asthma therapy.

• 한국치위생학회지
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• 제9권2호
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• pp.125-136
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• 2009

The purpose of this study was to investigate the recognition of the infection routes, symptoms and treatments of HBV by students of health-related departments so as to help students learn correct knowledge about hepatitis B and provide the basic data for establishment of oral health policies to prevent hepatitis B and improve the quality of infection management. For the subjects of this study, 666 students of health-related departments and other departments of universities in Daegu City, Gyeongbuk Province were arbitrarily chosen and given a questionnaire. Then the questionnaires collected between October 1st and 31st, 2007 were analyzed. Major findings from this study are summarized below. 1. Regarding general characteristics of the subjects, 311 were students of health-related departments and 355 were students of other departments. 55.9% of the health-related department students and 49.0% of the other department students received immunization against hepatitis. 36.0% of the health-related department students and 31.6% of the other department students had antibodies. 2. Regarding the recognition of the infection routes of HBV, the right answer "Infected through placenta" was chosen by more juniors(94.4%), sophomores(93.8%) and freshmen(74.1%) of health-related departments than other students in this order (P<0.05). The answer "Infected through sexual intercourse" was chosen by the highest percentage(75.0%) of juniors followed by freshmen(69.2%) and sophomores(31.9%) (P<0.05). 3. The percentages of health-related department students who knew that "HBV can develop into hepatic cirrhosis or liver cirrhosis were the highest among juniors(88.9%), freshmen(87.7%) and sophomores(68.8%) in this order(P<0.05). Among the other department students, the percentages of right answers to the question "Acute HBV infection shows jaundice" were the highest among juniors(75.0%), sophomores(74.8%) and freshmen(58.7%)(P<0.05).

• 동의생리병리학회지
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• 제19권2호
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• pp.398-406
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• 2005

This study was performed to investigate the effect of oral administration of GSRE against the asthma. Asthma was induced to Balb/c mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the eosinophil number in the BALF. Concentrations of IL-4, IL-5 in BALF and splenocyte were assessed by ELISA, IgG and IgE from serum were calculated by same method. Concentration of IL-4 in splenocyte was significantly decreased in GSRE group compared with control group. Concentrations of IL-5 from BALF and splenocyte were significantly decreased in GSRE group compared with control group, respectively. Level of IgE in serum was significantly decreased in GSRE group compared with control group, but not IgG. We found that the effect of GSRE extract in asthma was implicated in reductions of IL-4, IL-5 released from Th2 cell, and decreses of IgE, from plasma cell. These findings suggest that GSRE extract can produce anti-asthmatic effect, which may play a role in allergen-induced asthma therapy.

• 대한본초학회지
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• 제29권4호
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• pp.35-44
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• 2014

Objectives : Evodia rutaecarpa (Juss.) Benth.(ER) and Chaenomeles sinensis Koehne (CS) have multiple applications and were known to have anti-inflammatory effects. In the current study, we investigated to clearly understand the mechanism of therapeutic role for CS, ER and their combination in CIA model mice. Methods : DBA/1OlaHsd mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with CS, ER and their combination once a day for 7 weeks. Cytokine production and gene expression were assessed during CIA (collagen-induced arthritis) model mice in knee joint, lymph node (LN) using ELISA and FACS analysis. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. Result : Oral administration of CS, ER and their combination (150 mg/kg) significantly suppressed the progression of CIA, and significantly suppressed the progression of CIA and inhibited the production of TNF-${\alpha}$ and IL-6 in serum. The erosion of cartilage was dramatically reduced in mouse knees after treatment with CS plus ER. Conclusion : These result suggest that CS plus ER significantly suppressed the progression of CIA and that this action was characterized by the decreased production of TNF-${\alpha}$, IL-6 and collagen II specific antibody in serum.

• 동의생리병리학회지
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• 제21권1호
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• pp.39-49
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• 2007

Rheumatoid arthritis is an autoimmune disease involving multiple joint. In order to access the suppressive effects of JTT on rheumatoid arthritis and it's effects on immune system we investigated whether JTT could suppress the disease progression of collagen-induced arthritis. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with DW, JTT (200 or 400 mg/kg) or methotrexate (MTX, 30 mg/kg) as a positive control. Oral administration of JTT significantly suppressed the progression of CIA, which extend is comparable to that of MTX. Histological examination reveled that JTT inhibited infiltration of inflammatory cells into affected paw joint and bone erosion and cartilage destruction were greatly reduced compared with control. Total cell number of spleen, lymph node and peripheral blood were significantly reduced. The absolute number of CD19$^+$, CD3$^+$/CD69$^+$, CD4$^+$/CD25$^+$ cell in spleen from JTT treated mice were significantly decreased. The absolute number of CD19$^+$, CD3$^+$, CD3$^+$/CD69$^+$, CD4$^+$, CD4$^+$/CD25$^+$ CD8$^+$, CD49b, CD3/CD49b cells in draining lymph node were significantly increased compared with control. In peripheral blood mononuclear cells of JTT treated mice, the absolute number of CD4$^+$, CD4$^+$/CD25$^+$, CD3$^+$/CD69$^+$ cells were significantly decreased compared with control, while that of CD49b$^+$ was slightly increased. Infiltration of CD3$^+$ cells and CD11b$^+$/Gr-1$^+$ cells into paw joint was significantly reduced in JTT treated mice. The levels of pathologic cytokines including TNF-a and IL-6 in serum were significantly decreased by oral treatment with JTT The levels of IFN-g in the culture supernatant of splenocyte stimulated with CD3$^+$/CD28$^+$ or collagen were dramatically decreased, while the levels of IL-4 was increased under CD3$^+$/CD28$^+$ or collagen stimulation. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of JTT treated mice than control. Taken together, JTT has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use anti-rheumatic arthritic agent in human.