• Journal of Microbiology and Biotechnology
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• 제22권6호
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• pp.856-865
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• 2012

We describe the construction and immunobiological properties of a novel whooping cough vaccine candidate, in which the aroQ gene, encoding 3-dehydroquinase, was deleted by insertional inactivation using the kanamycin resistance gene cassette and allelic exchange using a Bordetella suicide vector. The aroQ B. pertussis mutant required supplementation of media to grow but failed to grow on an unsupplemented medium. The aroQ B. pertussis mutant was undetectable in the trachea and lungs of mice at days 6 and 12 post-infection, respectively. Antigen-specific antibody isotypes IgG1 and IgG2a, were produced, and cell-mediated immunity [CMI], using interleukin-2 and interferon-gamma as indirect indicators, was induced in mice vaccinated with the aroQ B. pertussis vaccine candidate, which were substantially enhanced upon second exposure to virulent B. pertussis. Interleukin-12 was also produced in the aroQ B. pertussis-vaccinated mice. On the other hand, neither IgG2a nor CMI-indicator cytokines were produced in DTaP-vaccinated mice, although the CMI-indicator cytokines became detectable post-challenge with virulent B. pertussis. Intranasal immunization with one dose of the aroQ B. pertussis mutant protected vaccinated mice against an intranasal challenge infection, with no pathogen being detected in the lungs of immunized mice by day 7 post-challenge. B. pertussis aroQ thus constitutes a safe, non-reverting, metabolite-deficient vaccine candidate that induces both humoral and cell-mediated immune responses with potential for use as a single-dose vaccine in adolescents and adults, in the first instance, with a view to disrupting the transmission cycle of whooping cough to infants and the community.

• Fisheries and Aquatic Sciences
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• 제25권4호
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• pp.243-249
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• 2022

Bacillus SW1-1 is a probiotic isolated from shrimp intestines. We investigated the effects of Bacillus SW1-1 coated diets on the growth, feed utilization, innate immunity, hematological parameters and resistance to Edwardsiella tarda in olive flounder (Paralichthys olivaceus). A commercial diet was used as the control (AP0) and two other diets were prepared by coating 0.25% (AP25) or 0.50% (AP50) probiotic powder which contains 1.0 × 10⁷ CFU/g Bacillus SW1-1. Four replicate groups of olive flounder (153 ± 2 g) were fed one of the diets for 12 weeks. Growth performance and feed utilization of the fish were not significantly affected by the dietary Bacillus SW1-1. After the challenge with E. tarda, AP50 group showed significantly higher survival than AP0 and AP25 groups. Innate immunity and anti-oxidant capacity of the fish were not significantly affected after the feeding trial. However, after the E. tarda challenge, the innate immune parameters (immunoglobulin, lysozyme and anti-protease) were significantly improved in fish fed AP25 and AP50 diets compared to those in fish fed AP0 diet. After the challenge test, significantly lower glucose level was observed in AP50 group compared to AP0 group. These results indicate that dietary supplementation of Bacillus SW1-1 could increase the disease resistance of olive flounder against E. tarda infection. The optimum coating levels of Bacillus SW1-1 needs to be further elucidated.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1585-1591
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• 2010

A comprehensive study on the production of inflammatory mediators in the lungs of BALB/c mice following infection with Stenotrophomonas maltophilia was conducted. The levels of pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-${\alpha}$), and interleukin-1${\beta}$ (IL-1${\beta}$) were raised in the lungs of infected mice compared with control. The production of anti-inflammatory cytokine IL-10 was slightly delayed. Its peak level was on the $2^{nd}$ day, whereas the peak of pro-inflammatory cytokines was observed on day 1 after intranasal challenge. This was accompanied by a rise in myeloperoxidase (MPO) and malondialdehyde (MDA) on day 1. The increase in MPO levels matched with histopathological observations, as neutrophils infiltration was detected on the first day. Alveolar macrophages (AMs) obtained from infected animals showed a higher rate of uptake and killing when exposed to bacteria in vitro, compared with similar experiments conducted with AMs from normal mice (control). This suggests that AMs were more efficient in cleaning the bacteria. The nitric oxide (NO) production however started early during infection but reached its maximum on the $3^{rd}$ day. No mortality was observed among the infected animals, and infection was resolved by the $5^{th}$ day post infection. No drastic changes in the lung tissue were observed on histopathological examination.

• 대한수의학회지
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• 제11권1호
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• pp.1-39
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• 1971

The purpose of present study was to investigate the possibility to immunize guineapigs and swine against Metastrongylus apri infection by the administration of irradiated infective larvae. Four main experiments were undertaken. Firstly, three groups of infective larvae irradiated at $3{\times}10^4r$, $4{\times}10^4r$, and $5{\times}10^4r$ respectively were inoculated to guineapigs and their immunogenic effects were examined from the clinical, anatomical, and serological viewpoints to decide the optimal dose of X-ray for the atenuation of them. Secondly, the migratory behavior of the larvae irradiated at the optimal dose was compared with that of normal infective larvae. Thirdly, pigs were inoculated with each 5,000 infective larvae irradiated at the optimal dose and the clinical, anatomical and serological responses of them before and after challenge were examined. Fourthly, the heated extract of adult M. apri which had been used as an antigen in the serological examinations was analyzed and compared with that of adult Ascaris suis and of adult Trichuris suis by immunoelectrophoretic method. The results obtained are summerized as follows: 1) The optimal dose of X-ray for the atenuation of the infective larvae which can minimize the pathogenecity but keep the antigenecity of the infective larvae was $5{\times}10^4r$. 2) Guineapigs could become completely resistant to subsequent challenge infection by the administration of 1,000 infective larvae irradiated at $5{\times}10^4r$, without showing any symptom of disease before and after the challenge. 3) There were some indication that guineapigs could acquire complete immunity after they overcome the infection with normal infective larvae. 4) It was shown that, in guineapigs, the $5{\times}10^4r$-irradiated larvae can migrate to the large intestine and mesenteric lymph node within a day, where they stay for as long as 16 days to stimulate the host's immunity. 5) It also was shown that, in guineapigs, the normal infective larvae challenged to resistant guineapigs can migrate to the large intestine and mesenteric lymph node, where they are affected by the immune mechanism of host within 10 days without further migration. 6) Pigs could become partially resistant to subsequant challenge by the administration of 5,000 infective larve irradiated at $5{\times}10^4r$; no clinical symptom occurred after the administration, but milder symptoms of parasitic bronchitis were observable after the challenge infection and fewer number of worms were detected from the lungs at autopsy compared with severe symptoms and much number of worms in control pigs. 7) It was shown that, in pigs, a few of the $5{\times}10^4r$-irradiated larvae can migrate to the lungs, where they stay for as long as 104 days in stunted and sterile states; their body-lengths were short and their uteri developted no eggs. 8) There was evidence that the male larvae were more susceptible to X-ray than the female larvae. 9) Antibodies relating to the administration with $5{\times}10^4r$-irradiated or normal larvae were detected from the sera of both guineapigs and pigs by means of indirect haemagglutination and agar diffusion precipitin tests. Relatively higher antibody titers were recorded by the former test, but precipitin bands were demonstrable only when the positive sera were concentrated in one tenth of original volume in the later one. 10) The antibody titers of pig sera began to rose on 14 days, kept their peak during the period from 14 th day to 21st day, and fell to a low level on 28 days after the administration of $5{\times}10^4r$ or normal infective larvae. 11) A slight increase in gamma globublin of the pig sera occurred following the administration. The gamma globulin level showed a tendency to fluctuate in acordance with the antibody level. 12) A marked eosionophilia occurred in pigs on 7 or 14 days following the administration. The eosinophil count showed the same tendency to fluctuate as the gamma globulin did. 13) It was shown that the serum antibodies detected by the heated extract of adult Metastrongylus apri react crossly with the heated extract of adult Ascaris suis but not with that of adult Trichuris suis in indirect haemagglutination and agar diffusion preciption reactions. 14) The heated extract of adult Metastrongylus apri could he divided into 9 antigenic components by immunoelectrophoresis, one (arc 4) of which was shown to be common to both extracts of adult Ascaris suis and adult Trichuris suis, and the other one (arc 9) to only the extract of adult Ascaris suis.

• Parasites, Hosts and Diseases
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• 제45권4호
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• pp.255-266
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• 2007

The protective effect of the Synadenium carinatum latex lectin (ScLL), and the possibility of using it as an adjuvant in murine model of vaccination against American cutaneous leishmaniasis, were evaluated. BALB/c mice were immunized with the lectin ScLL (10, 50, 100$[\mu}g$/animal) separately or in association with the soluble Leishmania amazonensis antigen (SLA). After a challenge infection with $10^6$ promastigotes, the injury progression was monitored weekly by measuring the footpad swelling for 10 weeks. ScLL appeared to be capable of conferring partial protection to the animals, being most evident when ScLL was used in concentrations of 50 and 100${\mu}g$/animal. Also the parasite load in the interior of macrophages showed significant reduction (61.7%) when compared to the control group. With regard to the cellular response, ScLL 50 and 100 ${\mu}g$/animal stimulated the delayed-type hypersensitivity (DTH) reaction significantly (P < 0.05) higher than SLA or SLA plus ScLL 10 weeks after the challenge infection. The detection of high levels of IgG2a and the expression of mRNA cytokines, such as IFN-$\gamma$, IL-12, and TNF-$\alpha$ (Th1 profiles), corroborated the protective role of this lectin against cutaneous leishmaniasis. This is the first report of the ScLL effect on leishmaniasis and shows a promising role for ScLL to be explored in other experimental models for treatment of leishmaniasis.

• 한국발생생물학회지:발생과생식
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• 제21권4호
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• pp.371-378
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• 2017

Interferon-stimulated gene 15 (ISG15) is known to interfere with viral replication and infection by limiting the viral infection of cells. Interferon-stimulated gene 15 (ISG15) interferes with viral replication and infectivity by limiting viral infection in cells. It also plays an important role in the immune response. In this study, tissue-specific expression of ISG15 in healthy rock bream samples and spatial and temporal expression analysis of rock bream ISG15 (RbISG15) were performed following rock bream iridovirus (RSIV) infection. RbISG15 expression was significantly higher in the eye, gill, intestine, kidney, liver, muscle, spleen, and stomach, but low in the brain. There were particularly high levels of expression in the liver and muscle. RbISG15 expression was also examined in several tissues and at various times following RSIV infection. ISG15 expression increased within 3 h in the whole body and decreased at 24 h after infection. In addition, temporal expression of several tissues following RSIV infection showed a similar pattern in the muscle, kidney, and spleen, increasing at 3 h and decreasing at 72 h. These results suggest that ISG15 plays an important role in the immune response of rock bream. Overall, this study characterizes the response of RbISG15 following RSIV infection.

• The Plant Pathology Journal
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• 제33권1호
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• pp.80-86
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• 2017

One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.

• Journal of Microbiology
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• 제39권3호
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• pp.186-194
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• 2001

Scrub typhus, caused by Orientia tsutsugamushi infection, is clinically and histopathologically characterized by local as well as systemic inflammatory reactions, indicating that orientiae induce mechanisms that amplify the inflammatory response. To reveal underlying mechanisms of chemoattraction and activation of responding leukocytes, expression of chemokine and tumor necrosis factor alpha (TNF-$\alpha$) genes in murine peritoneal macrophages after infection with the obligate intracellular bacterium Ο.tsutsugamushi was investigated. The genes that were unregulated included macrophage inflammatory proteins l$\alpha$/$\beta$(MIP-l$\alpha$/$\beta$), MIP-2, monocyte chemoattractant protein 1(MCP-1), RANTES (regulated upon activation, normal T-cell expressed and secreted), gamma-interferon-inducible protein 10(IP-10) and TNF-$\alpha$. Peak expression of these chemokines and TNF-$\alpha$ was observed between 1 and 3 h after infection. These responses returned to or approached baseline preinfection levels 6 h after challenge. Semiquantitative reverse transcription (RT)-PCR analysis revealed dramatic Increases during infection in the steady-state levels of mRNA ceding for the inhibitory subunit of NF-kB (IkB$\alpha$), whose transcription is enhanced by binding of NF-kB within the IkB$\alpha$promoter region. Thus, Ο. tsutsugamushi appears to be a stung inducer of chemokines and TNF-$\alpha$ which may significantly contribute to inflammation and tissue damage observed in scrub typhus by attracting and activating phagocytic leukocytes.

• IMMUNE NETWORK
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• 제9권2호
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• pp.46-52
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• 2009

Although tuberculosis poses a significant health threat to the global population, it is a challenge to develop new and effective therapeutic strategies. Nitric oxide (NO) and inducible NO synthase (iNOS) are important in innate immune responses to various intracellular bacterial infections, including mycobacterial infections. It is generally recognized that reactive nitrogen intermediates play an effective role in host defense mechanisms against tuberculosis. In a murine model of tuberculosis, NO plays a crucial role in antimycobacterial activity; however, it is controversial whether NO is critically involved in host defense against Mycobacterium tuberculosis in humans. Here, we review the roles of NO in host defense against murine and human tuberculosis. We also discuss the specific roles of NO in the central nervous system and lung epithelial cells during mycobacterial infection. A greater understanding of these defense mechanisms in human tuberculosis will aid in the development of new strategies for the treatment of disease.

• Parasites, Hosts and Diseases
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• 제37권2호
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• pp.101-107
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• 1999

Rats were immunized through an initial infection with 1,000 filariform larvae (L3) of Nippostrongylus brasiliensis and after complete expulsion of worms they were challenged with 1,000 L3 of Strongyloides venezuelensis to investigate whether cross-resistance developed against a heterologous parasite. Nippostrongylus brasiliensis immunized rats developed a partial cross-resistance against S.venezuelensis migrating larvae (MSL3) in the lungs and adult worms in the small intestine. The population of MSL3 in the lungs were significantly lower (p<0.05) in immunized rats($22.0{\;}{\pm}{\;}7.4$) compared with controls ($105.0{\;}{\pm}{\;}27.6$). The populations of adult worms, egg output and fecundity were initially decreased but from day 14 post-challenge they did not show any significant difference between immunized and control rats. However, the length of worm in immunized rat was revealed as retardation. Peripheral blood eosinophilia was significantly decreased (P<0.05) on day 7 post-challenge and then gradually increased which peaked on da 42 post-challenge when most of the worms were expelled. these results suggest that peripheral blood eosinophilia is strongly involved in the worm establishment and expulsion mechanisms.