• Journal of Periodontal and Implant Science
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• v.45 no.1
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• pp.14-22
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• 2015

Purpose: The purpose of this study was to compare serum amyloid A (SAA) protein levels with high-sensitive C-reactive protein (hs-CRP) levels as markers of systemic inflammation in patients with chronic periodontitis. The association of serum titers of antibodies to periodontal microbiota and SAA/hs-CRP levels in periodontitis patients was also studied. Methods: A total of 110 individuals were included in this study. Patients were assessed for levels of hs-CRP and SAA. Nonfasting blood samples were collected from participants at the time of clinical examination. The diagnosis of adipose tissue disorders was made according to previously defined criteria. To determine SAA levels, a sandwich enzyme-linked immunosorbent assay was utilized. Paper points were transferred to a sterile tube to obtain a pool of samples for polymerase chain reaction processing and the identification of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia. The serum level of IgG1 and IgG2 antibodies to P. gingivalis, A. actinomycetemcomitans, and T. forsythia was also determined. Results: SAA and hs-CRP levels were higher in periodontitis patients than in controls (P<0.05). In bivariate analysis, high levels of hs-CRP (>3 mg/L) and SAA (>10 mg/L) were significantly associated with chronic periodontitis (P=0.004). The Spearman correlation analysis between acute-phase proteins showed that SAA positively correlated with hs-CRP (r=0.218, P=0.02). In the adjusted model, chronic periodontitis was associated with high levels of SAA (odds ratio [OR], 5.5; 95% confidence interval [CI], 1.6-18.2; P=0.005) and elevated hs-CRP levels (OR, 6.1, 95% CI, 1.6-23.6; P=0.008). Increased levels of serum IgG2 antibodies to P. gingivalis were associated with high levels of SAA (OR, 3.6; 95% CI, 1.4-8.5; P=0.005) and high concentrations of hs-CRP (OR, 4.3; 95% CI, 1.9-9.8; P<0.001). Conclusions: SAA and hs-CRP concentrations in patients with chronic periodontitis are comparably elevated. High serum titers of antibodies to P. gingivalis and the presence of periodontal disease are independently related to high SAA and hs-CRP levels.

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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• pp.181-189
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• 1986

To apply ELISA to serodiagnosis of leptospirosis with killed whole cells from Leptospira interrogans serovars mwogolo (Mwogolo), copenhageni (M-20), WH-20, autumnalis (Akiyami A), cynopteri (3522 C), australis (Bacillico) and Leptospira biflexa serovar patoc (patoc 1), sensitivity and specificity was evaluated. The reactivity of IgM and IgG antibody in the sera from patients with leptospirosis, hemorrhagic fever with renal syndrome and other febrile disease and normal healthy control to the killed whole cells was analysed. The results were summarized as follows. 1. The reactivity (absorbance at 492mn) of IgM and IgG to L. mwogolo antigen in the sera of pattients with leptospirosis were $1.414{\pm}0.370$, $1.242{\pm}9.554$ respectively: hemorrhagic fever with renal syndrome, $0.329{\pm}0.131$, $0.239{\pm}0.126$; other febrile disease, $0.196{\pm}0.071$, $0355{\pm}0.141$; normal healthy control, $0.136{\pm}0.016$, $0.208{\pm}0.077$. 2. The reactivity of IgM and IgG to L. copenhageni, WH-20, L. autumnalis, L. cynopteri and L. anstralis antigens were similar to that to L. mwogolo antigen, but that to L. biflexa antigen was not discriminated among above disease. 3. Correlation coefficient between the MAT titer and ELISA OD (IgM) to the above antigens was in the range of 0.071-0.518. 4. As absorbance above 0.60 was determined positive for the diagnosis of leptospirosis, the sensitivity and specificity of IgG was 25-89% and 91-96% respectively. And those of IgM was 98-100% and 89-100% except L. biflexa (29%) respectively.

• Korean Journal of Veterinary Research
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• v.11 no.2
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• pp.123-136
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• 1971

(1) The non-specific inhibitors (NSI) in normal chicken sera were active against all the tested group A and group B arboviruses, but the group B arbovirus were more sensitive than group A arboviruses. (2) The titres of the NSI were distributed nearly uniformly among chickens from seven different age groups to group A arboviruses. In contrast, the NSI titres to group A arboviruses were found to increase with age. (3) No significant difference could be demonstrated between acetone-ether extraction and kaolin adsorption for removal of the NSI in normal chicken sera. (4) After heating, the NSI titres in chicken sera were increased for both group A and group B arboviruses. (5) After heating the sera at $80^{\circ}C$ and $100^{\circ}C$, kaolin adsorption was less efficient for removing the NSI than it, was in unheated serum. Acetone-ether extraction of the NSI was unimpaired after heating at $80^{\circ}C$ but was less efficient after heating at $100^{\circ}C$. (6) The NSI activity was found mainly in the first peak (IgM) and diffused to a part of second peak (IgG) by fractionation of chicken serum by gel filtration through Sephadex G200. After zonal centrifugation of chicken serum in a linear ten to 40 percent sucrose gradient all of the NSI activities were found on the top of the centrifugal tubes. These properties of large molecular size and low density indicated that the NSI in chicken serum were probably lipoproteins. (7) The natural agglutinins for goose erythrocytes in chicken sera were partially destroyed by acetone-ether extraction but not by kaolin adsorption, and were efficiently adsorbed with ten percent goose erythrocytes. No difference of the NA titre was demonstrated with diluents of different pH. (8) The NA in chicken serum was found to possess the properties of IgM by gel filtration through Sephadex G200 and zonal centrifugation in linear ten to 40 percent sucrose gradient.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.249-258
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• 1994

Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

• Toxicological Research
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• v.22 no.4
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• pp.357-364
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• 2006

This study was undertaken to develop a reliable mice model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. BALB/C mice and NC/Nga mice were sensitized twice with $100{\mu}l$ of 1% 2,4-dinitrochlorobenzene (DNCB) or vehicle (acetone : olive oil=4:1 mixture) in a week and challenged twice with $100{\mu}l$ of 0.2% DNCB or the vehicle at the following week. Mice were sacrificed at 19 days following the second DNCB or vehicle challenge for NC/Nga mice and at 28 days following the second DNCB or vehicle challenge for BALB/c mice. Upregulation of plasma 1gE, a hallmark of atopic dermatitis occurrence, was evident in the plasma obtained 4 day after the second DNCB challenge from BALB/c mice (approximately 4-fold) and NC/Nga mice (approximately 6-fold) treated with DNCB in comparison with that of the vehicle treated-control mice, and remain higher $3{\sim}4$ week after the second challenge. Ratio of plasma IgG1 versus IgG2a concentration was significantly higher in the mice treated with DNCB than the control mice, which also implies the skewed type-2 reactivity in vivo. Ratio of interleukin-4 versus interferon gamma produced in the splenic T cell culture supernatants was approximately 3-fold higher in the both strains of mice treated with DNCB than their control mice, respectively. The DNCB-treated mice demonstrated atopic dermatitis-like skin legions characterized with erythma, scaling, and hemorrhage, which was not observed with the control mice. Scratching on face or dorsal area was significantly more frequent (approximately 25-fold) in the DNCB-treated mice than the control at next day of the second DNCB challenge, and scratching frequency remains higher (approximately 4-fold) in the mice treated with DNCB than the control at 14 day following the second DNCB challenge. Overall, the mice model developed through sensitization and challenge with DNCB may be useful for research on atopic dermatitis and development of treatment materials for atopic dermatitis.

• Korean Journal of Community Nutrition
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• v.4 no.2
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• pp.132-138
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• 1999

The purpose of this study was to determine biochemical parameters related to protein and immunity. Subjects were 125 preschool children(M:69, F:56) residing in low income area of Seoul. Mean serum total protein of the children aged 6 was 7.3g/dl which was significantly higher than 6.6g/dl of the group aged 3. The mean serum albumin was 4.7g/dl for 3, 4, 5 age group, and 4.9g/dl for 6 age group and there was no significant difference. Serum retinol binding protein(RBP) is used as a sensitive indicatior of protein, because it tends to fall rapidly in response to protein status and respond to quickly dietary treatment. Mean RBP for each group(3, 4, 5 and 6 age group) were $2.5\mu{g}$/dl, $2.9\mu{g}$/dl, $2.7\mu{g}$/dl, $3.0\mu{g}$/dl. The Proportion of children whose RBP was than $2.6\mu{g}$/dl was 15.9%, 19.2%, 24.3% and 16.7%, respectively. The 24-hour urinary excretion of hydroxyproline was 7.9mg, 14.6mg, 11.7mg and 11.8mg for each group and the mean excretion of all children was 12.2mg/day. Children aged 3 were excreting significantly lower amount of hydroxprolinc per day than the children aged 4. The mean hydroxyproline index were 2.18, 2.39, 2.52, 2.80 for each age group and the mean of a group aged 6 was significantly higher than that of the group aged 3. The proportion of children assessed as malnourished and impaired growth(hydroxyproline index <2.0) was 18.8%, 4.9%, 2.5% and 4.3%, respectively. The nutrients which showed significant relationship with protein and immunity parameters were niacin, vitamin C and calcium. Vitamin C showed significant positive relationship(p<0.05-p<0.01) with serum RBP, total protein and globulin. The triceps skinfold thickness was significantly and positively correlated with serum globulin. Serum IgG showed significant positive relationship with height, weight, girth of chest and midarm circumference(p<0.05-p<0.01).

• 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.

• Research in Plant Disease
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• v.25 no.3
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• pp.131-135
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• 2019

Cylindrocarpon destructans causes ginseng root rot and produces radicicol that has an antifungal effect. In this study, we developed a method to detect this fungus using enzyme-linked immunosorbent assay (ELISA). Secreted proteins of C. destructans were used as antigens to obtain C. destructans-specific IgG from mouse. Out of 318 monoclonal antibodies generated from mouse, two antibodies (Cd7-2-2 and Cd7-2-10) showed highest specificity and sensitivity. Indirect ELISA using both antigens successfully detected C. destructans in soils, but direct ELISA using IgG conjugated with horseradish peroxidase failed to detect antigens in soils. The indirect ELISA developed here can efficiently detect the fungus and help manage ginseng root rot disease in fields.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• Pediatric Infection and Vaccine
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• v.4 no.1
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• pp.126-132
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• 1997

Measles outbreak in the world was decreased since measles vaccine had been introduced. Although vaccination rate is high, measles was not eradicated and measles reappeared among vaccinated children. We measured measles-specific antibody from the vaccinated and unvaccinated groups who had experienced apparent measles in the Seongnam city in 1993. The results were as follows. 1) The data included total 126 children (M:F=1 : 1). Age distribution of measles outbreak revealed 6 children in 5yr, 11 in 6yr, 20 in 7yr, 39 in 8yr, 22 in 9yr, 11 in 10yr, 11 in 11yr, and 6 in 12yr. 2) MMR vaccination rate was 78.6%(99/126) in the children who had experienced measles. Positive rate of measles-specific IgM Ab was 80.8% (80/99) among the vaccinated group and among 9E.6.% (25/27) the unvaccinated. 3) Positive rate of measles-specific IgG Ab was 90.9% (90/99) among MMR-vaccinated group, and 85.2% (23/27) in unvaccinated group. In conclusion, measles-specific IgM antibody have been detected more than 1 month in most patients. The relatively high proportion of measles-specific IgM positivity may mean primary vaccine failure. To booster the antibody titers and to prevent measles epidemic in school-aged children, revaccination of measles should be considered.