• Journal of Veterinary Science
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• v.23 no.3
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• pp.41.1-41.15
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• 2022

Background: Proliferative enteritis caused by Lawsonia intracellularis undermines the economic stability of the swine industry worldwide. The development of cost-effective animal models to study the pathophysiology of the disease will help develop strategies to counter this bacterium. Objectives: This study focused on establishing a model of gastrointestinal (GI) infection of L. intracellularis in C57BL/6 mice to evaluate the disease progression and lesions of proliferative enteropathy (PE) in murine GI tissue. Methods: We assessed the murine mucosal and cell-mediated immune responses generated in response to inoculation with L. intracellularis. Results: The mice developed characteristic lesions of the disease and shed L. intracellularis in the feces following oral inoculation with 5 × 10⁷ bacteria. An increase in L. intracellularis 16s rRNA and groEL copies in the intestine of infected mice indicated intestinal dissemination of the bacteria. The C57BL/6 mice appeared capable of modulating humoral and cell-mediated immune responses to L. intracellularis infection. Notably, the expression of genes for the vitamin B12 receptor and for secreted and membrane-bound mucins were downregulated in L. intracellularis -infected mice. Furthermore, L. intracellularis colonization of the mouse intestine was confirmed by the immunohistochemistry and western blot analyses. Conclusions: This is the first study demonstrating the contributions of bacterial chaperonin and host nutrient genes to PE using an immunocompetent mouse model. This mouse infection model may serve as a platform from which to study L. intracellularis infection and develop potential vaccination and therapeutic strategies to treat PE.

• Journal of Environmental Health Sciences
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• v.21 no.3
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• pp.16-22
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• 1995

This study was designed to investigate the antibody production to sheep red blood cells(SRBC) and proliferation of mitogen-stimulated spleen cells in Balb/c mice which received cadmium chloride. The mice were divided into three independent groups which were one control and two experimental groups by the cadmium treatment or not. No specific treatment was done for the control group. One of two experimental groups, which is called 'pre-treatment group' in this paper, was subcutaneously injected with low dose of cadmium chloride(0.5 mg/kg/day) for 5 consecutive days before the primary SRBC immunization. The other called 'non-pretreatment group' was only pretreated with normal saline. Both experimental groups were intraperitoneally injected with high dose of cadmium chloride(5 mg/kg) 8 hours before the primary immunization. Mice were intraperitoneally immunized twice with 2% SRBC suspension containing $10^8$ cells. The results obtained were as follows, 1. The PFG responses to SRBC were significantly increased in two experimental groups, cadmium pretreatment and non-pretreatment compared with that of control group(p<0.05). 2. The total antibody titers to SRBC in cadmium treated groups were similar to that of control group, but titers of IgG antibody were significantly elevated(p<0.01). 3. The proliferation response of spleen lymphocytes to various mitogens was suppressed in proportion to the concentration of cadmium and the degree of cadmium accumulation in liver was increased in the cadmium treated groups. These results suggest that cadmium chloride could affect on mouse immune response, especially its cell mediated immune response could be decreased while its humoral immune response could be increased, which may not be influenced by the administration methods or pretreatment of cadmium to mouse.

• Journal of Preventive Medicine and Public Health
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• v.28 no.1 s.49
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• pp.27-42
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• 1995

The in vivo and in vitro humoral and cell-mediated immune responses of lymphocytes of BALB/c mouse exposed to mercury chloride$(HgCl_2)$ were investigated. In vitro exposure of the splenocytes to mercury chloride produced overt cytotoxicity in 3 hours period. The $IC_{50}$(the concentration required to inhibit a splenocyte viability by 50%) for mercury chloride was >0.1mM for cytotoxicity. In vivo mercury chloride exposed mice were significantly depressed delayed type hypersensitivity(DTH) response to sheep red blood cells(SRBC) in a dose-dependent manner compared with control group. Mercury chloride inhibited the proliferative responses of splenocytes to lipopolysaccharide, pokeweed mitogen, concanavalin A and phytohemagglutinin in a dose-dependent manner. Hemagglutinin response to SRBC in mercury chloride exposed mice was significantly depressed in a dose-dependent manner compared with control group. After 7 weeks of mercury chloride exposure in vivo, mercury chloride induced an increase of nonspecific serum $IgG_1$ and IgE levels in BALB/c mice.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2003.06a
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• pp.55-62
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• 2003

The mucosal immune system provides a first line of defense against invasion of infectious agents via inhalation, ingestion and sexual contact. For the induction of protective immunity at these invasion sites, one must consider the use of the CMIS, which interconnects inductive tissues, including PP and NALT, and effector tissues of the intestinal, respiratory and genitourinary tracts. In order for the CMIS to induce maximal protective mucosal immunity, co-administration of mucosal adjuvant or use of mucosal antigen delivery vehicle has been shown to be essential. When vaccine antigen is administered via oral or nasal route, antigen-specific Th 1 and Th2 cells, cytotoxic T lymphocytes(CTLs) and IgA B cell responses are effectively induced by the CMIS. In the early stages of induction of mucosal immune response, the uptake of orally or nasally administered antigens is achieved through a unique set of antigen-sampling cells, M cells located in follicle-associated epithelium(FAE) of inductive sites. After successful uptake, the antigens are immediately processed and presented by the underlying DCs for the generation of antigen-specific T cells and IgA committed B cells. These antigen-specific lymphocytes are then home to the distant mucosal effector tissues for the induction of antigen-specific humoral(e.g., IgA) and cell-mediated (e.g., CTL and Th1) immune responses in order to form the first line of defense. Elucidation of the molecular/cellular characteristics of the immunological sequence of mucosal immune response beginning from the antigen sampling and processing/presentation by M cells and mucosal DCs followed by the effector phase with antigen-specific lymphocytes will greatly facilitate the design of a new generation of effective mucosal antigen-specific lymphocytes will greatly facilitate the design of a new generation of a new generation of effective mucosal adjuvants and of a vaccine deliver vehicle that maximizes the use of the CMIS.

• Nutritional Sciences
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• v.1 no.1
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• pp.22-28
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• 1998

Plasma concentrations of Vitamins E and A were measured in 15 non-insulin dependent Korean female subjects and 15 age-matched normal subjects using reversed-phase high-performance liquid chromatography. No differences were found in plasma Vitamin E concentrations between the 2 groups. Plasma Vitamin A concentrations were higher in subjects with non-insulin dependent diabetes melitus (NIDDM). The effects were evaluated of 4 weeks of daily supplementation of 400 mg Vitamin E on plasma levels of these two vitamins. In addition, the effects were observed for Vitamin E supplementation on oxidative stress and immune-related compound productions in non-insulin dependent diabetic patients and control subjects. After treatment with Vitamin E, plasma Vitamin E concentrations were significantly elevated in both groups. Basal plasma thiobarbituric acid reactive substances (TBABS) were identical, and a decreased level of TBARS caused by Vitamin E was observed only in the diabetic group (0.02739$\pm$0.0024 versus 0.01814$\pm$0.0008 nmols malondialdehyde equivalents/dl plasma ; p<0.05). The basal and after-treatment levels of immunoglobulins A, G, M were identical in control and diabetic groups, indicating that Vitamin E did not appear to alter gross humoral responses in this study. However, elevation of Complement 3 ($C_3$) was noticed due to Vitamin E supplementation, revealing a possible effect of vitamin E on one aspect of humoral immunity, Furthermore, an increase in prostaglandin E_2 ($PGE_2$) levels in diabetic patients was normalized by Vitamin E supplementation. This suggests indirectly that the depressed cell-mediated response due to elevated $PGE_2$ could be normalized. For the definitive antioxidant intake recommendations for prevention and treatment of adverse effects of non-insulin dependent diabetes, evidence from intervention trials like this study should be collected. The present data suggests that Vitamin E may oxen some protective effects against oxidative damage and might have beneficial effects of partial immune-stimulation.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.307-313
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• 1994

We have previously shown that crude water extract of Euonymus alatus (EA) had strong prophylactic effect against chemically induced-and tumor cell implanted-cancer, and that the mechanisms responsible for its antitumor effects were due to nonspecific enhancement of the NK cell activities and the cell mediated immunity. However, it was unknown that any components of crude extract did work so, since it consisted of several components. In this paper, we fractionated the crude watar EA-extract into several fraction such as hexane-, ethylether-, ethyl acetate-, n-butanol- and water soluble-fraction, and screened the immune regulating activities of each fraction by the evaluation of lymphokine production and activated lymphocyte proliferation. As a result of the component fraction of EA-extract, it was found that n-butanol fraction was a potent immunostimulator, and the remained water soluble fraction also contained some stimulator, But, other fraction did not showed any remarkable effect. It is therefore suggested that EA-glycosides in n-butanol fraction may be new one of the potent biological response modifiers. The present study was also undertaken in an efforts to investigate the effects of elm-bark(EB, Ulmus clavidiana var japonica), which has been used for curing ulcer and inflammation as a folk medicine without any kind of experimental evidence to support this, on the cellular- and humoral-immune responses, lymphocyte function and NK cell activities in mice. Regardless of time and duration of EB-treatment, Arthus reaction and antibody response to SRBC were not modified by EB, but delayed hypersensitivity to SRBC was significantly enhanced only when EB was treated prior to SRBC-sensitization. EB slightly inhibited the proliferation responses of splenocytes to PHA-stimulation, but it significantly augmented the responses of these cells to S aureus Cowan 1 and Con A-activation, and these effects were manifested only when EB was added at culture initiation. EB did not influence Ig secretion of spleen cells but it significantly augmented the Con A-induced 1L 2 and MIF production of splenocytes. NK cell activities of splenocytes were markedly riled when effector cells were pretreated with EB and this augmentation was dine to the increase of binding affinity of effector cells to target cells and the target cell lytic activities of effector cells. These results led to the conclusion that EB triggers increase of cellular immune responses, such as delayed hypersensitivitiy, lymphokine production and NK cell activities. Also these results suggested that EB contains potent immune stimulants, which may provide the rational basis for their therapeutic use as one of the new biological response modifiers.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.4
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• pp.560-563
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• 2004

The study was undertaken to evaluate the effect of naked neck gene on mortality, cell mediated and humoral immune response in white plumage broiler population. The mortality of homozygous naked neck (Na/Na) broilers (11.71%) was comparatively lower than that of heterozygous naked neck (Na/na) (12.28%) and normally feathered (na/na) (13.59%) broilers. The humoral immune response was measured against (1% v/v) sheep red blood cells (SRBC) for total haemagglutinin (HA) antibody, 2-mercaptoethanol resistance (MER) or (IgG) antibody and 2-mercaptoethanol sensitive (MES) or (IgM) antibody titre on 7 days post-immunization. The titre was expressed as log2 of the highest dilution which shows complete haemagglutination. Total HA titers of Na/Na and Na/na (11.05$\pm$0.53 and 11.09$\pm$0.38) were comparatively higher than that of na/na (10.26$\pm$0.42). The MES antibody titre of Na/Na (8.50$\pm$0.53) and Na/na (7.63$\pm$0.45) broilers were significantly higher as compared to na/na (6.11$\pm$0.32) broilers. The MER titre of na/na genetic group (4.15$\pm$0.42) was significantly higher than Na/Na (2.55$\pm$0.37) and comparatively higher than Na/na (3.45$\pm$0.38) broilers. In vivo cell response to phytohaemagglutinin-P (PHA-P), measured as Foot Index (FI) in mm expressed significantly higher response in Na/na (0.473$\pm$0.05) and Na/Na (0.413$\pm$0.04) broilers as compared to na/na (0.304$\pm$0.03) broilers. The result of present study suggested that white plumage naked neck broilers had better immune response as compared to normally feathered broilers.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1613-1619
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• 2016

Francisella tularensis is a highly virulent pathogen of humans and other mammals. Moreover, F. tularensis has been designated a category A biothreat agent, and there is growing interest in the development of a protective vaccine. In the present study, we determine the in vitro and in vivo immune responses of a subunit vaccine composed of recombinant peptides Tul4 and FopA from epitopes of the F. tularensis outer membrane proteins. The recombinant peptides with adjuvant CpG induced robust immunophenotypic change of dendritic cell (DC) maturation and secretion of inflammatory cytokines (IL-6, IL-12). In addition, the matured DCs enabled ex vivo proliferation of naive splenocytes in a mixed lymphocyte reaction. Lastly, we determined the in vivo immune response by assessment of antibody production in C57BL/6 mice. Total IgG levels were produced after immunization and peaked in 6 weeks, and moreover, Tul4-specific IgG was confirmed in the mice receiving peptides with or without CpG. Based on these results, we concluded that the recombinant peptides Tul4 and FopA have immunogenicity and could be a safe subunit vaccine candidate approach against F. tularensis.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.190-196
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• 2016

In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.

• Clinical and Experimental Vaccine Research
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• v.12 no.2
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• pp.176-178
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• 2023

We report the case of a toddler, with a history of mild atopic dermatitis (AD) since early infancy, presented to the Giannina Gaslini, a pediatric polyclinic hospital, 14 days after measles-mumps-rubella (MMR) vaccination, for the occurrence of a disseminated vesico-pustular rash, accompanied by general malaise, fever, restlessness, and anorexia. Eczema herpeticum (EH) was diagnosed clinically and confirmed by laboratory examinations. The exact pathogenesis of EH in AD is still debated and possibly involves an inter-play between altered cell-mediated and humoral immunity, failure to up-regulate antiviral proteins, and exposure of viral binding sites through the dermatitis and an epidermal barrier failure. We hypothesize that in this particular case, MMR vaccination might have played an additional important role in the alteration of innate immune response, facilitating the manifestation of herpes simplex virus type 1 in the form of EH.