• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.5
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• pp.705-711
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• 2018

Objective: This experiment was conducted to evaluate the effects of golden needle mushroom (Flammulina velutipes) stem waste (FVW), on organic eggs production, calcium utilization, antibody response, serum immunoglobulin, and serum cytokine concentration at early phase of production in laying hens. Methods: A total 210, 19 weeks old aged ISA Brown layers were randomly assigned into 5 equal treatment groups, with 7 replications of 6 hens each. Dietary treatment included a standard basal diet as control; antibiotic (0.05% flavomycin); 2% FVW; 4% FVW; and 6% FVW. The experimental duration was 10 weeks. Results: There was no significant differences (p>0.05) on hen day egg production, egg weight, egg mass, feed intake, and feed conversion ratio (FCR) among experimental groups. Unmarketable eggs were significantly lower (p<0.05) both in 4% FVW and 6% FVW fed groups than control group. The calcium retention and calcium in egg shell deposition were significantly higher (p<0.05) in FVW inclusion groups than control and antibiotic groups. Antibody titers against Newcastle diseases were significantly higher (p<0.05) in 6% FVW fed group (except combined with 4% FVW at day 147) and infectious bronchitis were significantly higher (p<0.05) in FVW fed groups (except 2% FVW and 4% FVW at day 161) than control and antibiotic groups. Serum immunoglobulin sIgA was significantly higher (p<0.05) in all levels of FVW and IgG was significantly higher (p<0.05) in 4% FVW than control and antibiotic groups. Serum cytokine concentration interleukin-2 (IL-2) was significantly higher (p<0.05) in 6% FVW; IL-6 and tumor necrotic $factor-{\alpha}$ were significantly higher (p<0.05) both in 4% FVW and 6% FVW than control and antibiotic groups; IL-4 was significantly higher (p<0.05) in antibiotic, 2% FVW and 4% FVW fed groups than control. Conclusion: F. velutipes mushroom waste can be used as a novel substitute for antibiotic for organic egg production and sound health status in laying hens.

• The Journal of Dong Guk Oriental Medicine
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• v.9
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• pp.51-71
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• 2000

The present experiments were designed to study the influence of WalgookwhanhabBojoongikgitang on immune functions of Balb/c mice under stress condition. WalgookwhanhabBojoongikgitang was orally administered to the mice for 15 days. On the 10th day the mice were immunized with sheep red blood cells (SRBC) and then subjected to electric footshock for 5 days (2 sessions a day, 11 footshocks a 31 min-session). The immune responses to SRBC were determined by means of hemagglutination and BIT cell populations in the spleen were studied by FACS analysis on the 16th day. The results were as follows. 1. After electric footshock, IDlce became sluggish and crowded to one side of the cage. Increased antibody titer for SRBC, increased B cell population, and decreased T cell population in the spleen were also observed. These results confirm that electric footshock caused stress inducing immunological and behavioral changes in Balblc mice. 2. Walgookwhanhab-Bojoongikgitang administration significantly antagonized the effect of electric footshock on the antibody titer for SRBC. As a result, antibody titers for SRBC in the mice treated with Walgookwhanga-Bojoongikgitang were maintained at the similar levels as those of control group mice even after the electric foots hock. 3. Walgookwhanhab-Bojoongikgitang administration also antagonized the effect of electric footshock on the BIT populations in spleen. As a result, Band T populations in the mice treated with Walgookwhanhab-Bojoongikgitang were maintained at the similar levels as in the control group mice even after the electric footshock. Taken together, Walgookwhanhab-Bojoongikgitang seem to help Balb/c mIce to maintain their humoral immune response and immune cell populations at a normal range under the stress conditions, suggesting its possible therapeutic use as a immune function modulator.

• Journal of Animal Science and Technology
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• v.46 no.3
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• pp.495-502
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• 2004

Intimin, the product of eae gene in EHEC O157:H7, is required for intimate adherence. In this study, the C-terminaI region(281 amino acids) of the EHEC OI57:H7 intimin were expressed as a protein fusion with (His)$_6$ which was used to raise antiserum in rabbits. The antiserum reacted in western blot with a 94kDa outer membrane protein of EHEC O157:H7. It was observed that the antibody titers both in egg yolk and serum appeared in 2${\sim}$4 weeks after immunization with fusion protein. At the time of 8 weeks, the titre of egg yolk was found to be higher than that of sera. According to the results of neutralization test, chicken egg-yolk antibody(lgY) against the recombinant intimin strongly reacted to EHEC O157:H7. We conclude that a truncated recombinant intimin could be used as an immunogen to elicit antibody(lgY) against O157:H7.

• Korean Journal of Head & Neck Oncology
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• v.15 no.1
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• pp.46-51
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• 1999

Background: Total thyroidectomy and postoperative radiodiodine ablation therapy in differentiated thyroid carcinomas enhance the reliability of serum thyroglobulin(Tg) levels and radioiodine scan in detecting recurrence or distant metastasis. There have been, however, some limitations in using these methods under certain conditions. Recently, several reports have indicated that thyroid peroxidase(TPO) could be used as an alternative tumor marker. We aimed to estimate the significance of serum TPO levels in differentiated thyroid carcinoma. Materials and Methods: Forty-eight patients who had undergone total thyroidectomy due to papillary thyroid carcinomas and who had been followed-up for at least 3 years were classified into two groups: 27 patients without any evidence of recurrence in group 1; and 20 patients with recurrence or distant metastasis in group 2. All patients were examined by radioiodine scans. Serum Tg, TSH, antithyroglobulin antibody, and TPO were measured and the relationships were statistically analyzed. The sensitivity and specificity of $^{131}I$ scan, serum Tg, and serum TPO were evaluated. Results: Serum Tg levels were $3.81{\pm}5.16ng/mL$ in group 1 and $147.02{\pm}193.75ng/mL$ in group 2. Only 2 patients in group 1 showed Tg levels exceeding 10ng/mL. In contrast, 4 patients in group 2 were under 10ng/mL. Serum antithyroglobulin antibody and TSH levels showed no statistical difference between the two groups. In group 1, 16 patients showed negative serum TPO results, and 4 patients in group 2 showed negative results. There was no correlation among serum Tg levels, antithyroglobulin antibody titers, and serum TPO levels in each group. In group 2, 4 patients with negative serum Tg levels showed positive TPO results and positive whole body scans. Two cases with false negative $^{131}I$ scans showed positive serum TPO and Tg results. In 4 cases showing false negative serum TPO levels, serum Tg levels and $^{131}I$ scans were positive. Conclusion: Serum Tg levels, radioiodine scans, and serum TPO levels can be clinically used as complementary methods in the diagnosis of recurrent or metastatic thyroid carcinomas. Serum TPO levels may be helpful when other methods fail to detect recurrences or distant metastasis in highly suspected patients.

• Parasites, Hosts and Diseases
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• v.25 no.1
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• pp.1-6
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• 1987

This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.

• Journal of Periodontal and Implant Science
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• v.51 no.4
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• pp.226-238
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• 2021

Purpose: Although several reports have described the relationship between periodontal disease and cardiovascular disease, information about the association between periodontal disease and the progression of degenerative aortic stenosis (AS) is lacking. Therefore, we performed a retrospective, single-center, pilot study to provide insight into this potential association. Methods: Data from 45 consecutive patients (19 men; median age, 83 years) with mild or moderate degenerative aortic stenosis were analyzed for a mean observation period of 3.3±1.9 years. The total amount of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis and titers of serum immunoglobulin G (IgG) against periodontal bacteria and high-sensitivity C-reactive protein (hs-CRP) were evaluated. Aortic valve area (AVA), maximal velocity (Vmax), mean pressure gradient (mean PG), and the Doppler velocity index (DVI) were evaluated. The change in each parameter per year ([Parameter_LATEST-Parameter_BASELINE]/Follow-up Years) was calculated from the retrospective follow-up echocardiographic data (baseline vs. the most recently collected data [latest]). Results: No correlation was found between the concentration of periodontopathic bacteria in the saliva and AS status/progression. The anti-P. gingivalis antibody titer in the serum showed a significant positive correlation with AVA and DVI. Additionally, there was a negative correlation between the anti-P. gingivalis IgG antibody titer and mean PG. The hs-CRP concentration showed positive correlations with Vmax and mean PG. Meanwhile, a negative correlation was observed between the anti-P. gingivalis IgG antibody titer and ΔAVA/year and Δmean PG/year. The hs-CRP concentration showed positive correlations with Vmax and mean PG, and it was significantly higher in patients with rapid aortic stenosis progression (ΔAVA/year <-0.1) than in their counterparts. Conclusions: Our results suggest that periodontopathic bacteria such as A. actinomycetemcomitans and P. gingivalis are not directly related to the status/progression of degenerative AS. However, inflammation and a lower immune response may be associated with disease progression.

• Journal of agricultural medicine and community health
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• v.13 no.1
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• pp.41-59
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• 1988

In many of the helminthic infections, diagnosis is accomplished by the demonstration of the eggs or, occasionally the adult worms or their parts. Diagnosis can be made by the identification of larval stage obtained from stool or surgically extracted materials too. However some kinds of parasitic disease can not be diagnosed by above mentioned procedure alone. Brain cysticercosis, ectopic paragonimiasis, Capillaria hepatica infection in liver is a good example. In such a case, immunologic method would be helpful for the decision of physician. In this paper, immunologic tools such as indirect hemagglutination test, indirect fluorescent antibody test, circumoval precipitation test, ELISA, western blot were applied for the diagnosis of Clonorchis sinenisis, Cysticercosis and C. hepatica a infection and their efficacy was evaluated. The results obtained were as follows ; 1) In the diagnosis of clonorchiasis, ELISA revealed sensitiveity of 83.3%, but cross reaction against antibody of Paragonimus westermani and Taenia species were observed. For the identification of cross reaction and species specific band of Ag-Ab reaction, western blot was applied. 59Kd relative molecular weight and 21Kd band were identified as a Clonorchis sinensis specific band. OD values of ELISA performed with sera of 18 months after praziquantel treatment decreased to half level compared to that of before treatment. Negative conversion rate of ELISA after 18 months of treatment was 60%. 2) In the diagnosis of cysticercosis, IFAT disclosed 95.8%(23/24) of sensitivity and reaction was most strongly occurred in inner membrane. ELISA revealed 90.0% (36/40) of sensitivity, but cross reaction was observed in both technique. In western blot, 91, 63 and 21Kd Mw bands were identified as a strongly positive band. Among them 63Kd band showed positive reaction against almost all sera of cysticercosis patient. 3) Circumoval precipitation, ELISA, IFAT, showed 85.0% of sensitivity in the diagnosis of C. hepatica infection in rat. The antigenic localities were inner membrane of sectioned egg antigen on the prectipitates around the mucoid plugs which were induced by circumoval precipitation reaction. Sera from rats infected with 2000eggs were collected periodically to observe the changing patterns of antibody titers by IFAT and ELISA, which showed that high titers were detected at weeks 3 and 5, then gradually declined through weeks 9until to negatively converted at weeks 13.

• Clinical and Experimental Pediatrics
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• v.48 no.9
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• pp.969-975
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• 2005

Purpose : We performed a study of clinical findings of Mycoplasma Pneumonia in children, to know differences between recent clinical manifestations of Mycoplasma pneumonia and previous studies. Methods : The subjects of this study were 393 children who were diagnosed as Mycoplasma pneumonia with high titers of Mycoplasma antibody(${\geq}1$ : 160) or fourfold rises of Mycoplasma antibody at Chung Ang University Hospital from January 1998 to December 2003. We practiced a retrospective study on the clinical manifestations of Mycoplasma pneumonia based on their medical records. Results : Male to female ratio was 1.06 to 1 and mean age was $4.32{\pm}2.94years$. The highest incidence was in the age of 2 to 3 years(18.6 percent). Most frequent months were October, and November in 2000, April in 2002, and October and, December in 2003. Twenty six point seven percent showed allergic diseases. Second degree relatives of 10.7 percent patients had allergic diseases. Forty three point three percent were admitted before this admission for pneumonia. Allergic tests were positive in 65.7 percent. Cough, and sputum were the most common symptoms and abdominal pain, and vomiting were the most frequent extrapulmonary symptoms. Atelectasis and pleural effusion were seen in 2.5 percent and 1.8 percent. Infiltrations were more common on the right side. Titers of each simultaneous test for cold agglutinin and mycoplasma antibody were not in proportion to each other (P=0.163). Conclusion : The onset age of mycoplasma pneumonia was found to be lower than it used to be. More patients had a past medical history or a family history of allergic disease, and more wheezing was heard and associated with recurrent pneumonia.

• Journal of Life Science
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• v.30 no.10
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• pp.905-911
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• 2020

The adjuvant effect of PAMAM dendrimer G4 (PAMAM) on the induction of humoral and cellular immune responses against keyhole limpet hemocyanin (KLH) was examined. Mice were immunized subcutaneously twice at two-week intervals with KLH, with or without PAMAM dendrimer (100 ㎍/mouse), and the mice immunized with KLH+PAMAM showed significantly higher antibody titers against KLH than those immunized with KLH alone. The assay for determining the isotypes of the antibodies showed that PAMAM augmented the KLH-specific antibody titers of IgG1, IgG2a, IgG2b, IgG3, and IgM. In addition, mice immunized twice with KLH+PAMAM followed by a subcutaneous injection of KLH (20 ㎍/site) 7 weeks after the primary immunization exhibited a higher delayed-type hypersensitivity (DTH) reaction than those treated with KLH alone. In an in vitro analysis of T lymphocyte proliferation in response to KLH in week 8, the splenocytes of mice treated with KLH+PAMAM showed significantly higher proliferating activity than those treated with KLH alone, and the culture supernatants of cell cultures from mice immunized with added PAMAM dendrimer showed higher levels of KLH-specific cytokine (IL-4 and IFN-r) production. These results suggest that PAMAM dendrimer G4 possesses a potent immune-adjuvant activity for enhancing both humoral and cell-mediated immunity specific to foreign antigens.