• Parasites, Hosts and Diseases
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• 제46권4호
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• pp.279-283
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• 2008

To examine humoral immune responses in the host, we measured serum antibody levels in different strains of mice (ICR, BALB/c, and C3H) experimentally infected with Neodiplostomum seoulense. Specific IgG antibody levels were increased remarkably with little difference among 3 strains of mice infected with N. seoulense from day 7 to 35 post-infection. More target proteins of adult parasites reacted with IgG at the time when the worm recovery decreased compared with other times. More than 20 protein bands, from 14 kDa to 94 kDa in size, were separated from the crude antigen of N. seoulense adults by SDS-PAGE, and among them 26, 30, 35, 43, 54, 67, and 94 kDa proteins were the major antigenic proteins. The results suggest that significant IgG antibody responses occur against N. seoulense in mice and this may be related with expulsion of worms.

• 동아시아식생활학회지
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• 제9권2호
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• pp.200-206
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• 1999

Salmonella typhimurium 항원에 면역된 산란계의 계란항체(IgY)생산 및 증가를 측정한 결과, 산란계가 이들 항원에 면역반응을 일으켜 2주 후 혈청과 난황에서 정상군에 비하여 뚜렷한 항체가 생성능을 보였으며. 혈청내의 IgG생성에 비하여 계란항체(IgY)의 생성이 다소 낮은 것으로 나타났다. 혈청 및 난황내의 단백질 함량은 정상군에 비하여 약간 높은 경향을 나타내어 산란계의 혈청 및 난황내의 단백질이 이들 항원에 의하여 증가하는 것으로 나타났다. DEAE-Sephacel column을 통과한 분획은 두 개의 peak를 나타내었다. 각 분획의 IgY농도를 ELISA법으로 확인한 결과 시험관 350-635$m\ell$에 해당되는 두번째 peak에서 대부분 측정되었다. 분리 정제된 IgY의 분자량을 측정한 결과, heavy chain 72-75KD, light chain 30-40KD정도였다. 이상의 결과에서, 본 실험에서 사용된 IgY는 항원에 대한 특이성 및 생화학적 특성 등을 고려해 볼 때, 식품산업 소재, 의약품 소재, 연구용 kit와 진단용 kit 및 축산분야 등 여러 분야에 응용될 수 있으리라 기대된다.

• Parasites, Hosts and Diseases
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• 제59권3호
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• pp.257-263
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• 2021

Human gnathostomiasis is a parasitic disease caused by Gnathostoma nematode infection. A rapid, reliable, and practical immunoassay, named dot immuno-gold filtration assay (DIGFA), was developed to supporting clinical diagnosis of gnathostomiasis. The practical tool detected anti-Gnathostoma-specific IgG4 in human serum using crude extract of third-stage larvae as antigen. The result of the test was shown by anti-human IgG4 monoclonal antibody conjugated colloidal gold. The sensitivity and specificity of the test were both 100% for detection in human sera from patients with gnathostomiasis (13/13) and from healthy negative controls (50/50), respectively. Cross-reactivity with heterogonous serum samples from patients with other helminthiases ranged from 0 (trichinosis, paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis) to 25.0% (sparganosis), with an average of 6.3% (7/112). Moreover, specific IgG4 antibodies diminished at 6 months after treatment. This study showed that DIGFA for the detection of specific IgG4 in human sera could be a promising tool for the diagnosis of gnathostomiasis and useful for evaluating therapeutic effects.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.455-464
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• 1995

연구배경: 인체의 결핵균 감염에 대하여 체액성 면역과 세포 매개성 면역이 함께 관여하며 효소결합 면역 분석법으로 결핵균 항원에 대한 항체(IgG)를 측정하는 것은 결핵성 흉막염 진단법으로 이용할 수 있다. 방법: 1992년 5월부터 1994년 7월까지 인하대학교 인하병원에 흉막염으로 입원하였던 환자 중 원인이 확진된 결핵성 흉막염 환자 40예, 비결핵성 흉막염 환자 19예를 대상으로 하여 혈청 및 흉막액에서 PPD 항원과 LAM-B 항원에 대한 IgG 항체가의 흡광도를 측정 하였다. 결과: 1) 결핵성 흉막염군의 흉막액 및 혈청내 PPD와 LAM-B 항체가는 비결핵성 흉악염군보다 유의하게 높았다.(p<0.0005) 2) 혈청내 PPD와 LAM-B 항체가는 흉막액내 항체가 보다 높았다. 3) PPD와 LAM-B에 대한 흉막액 항체와 혈청 항체간에는 의미있는 상관관계를 보였다. 4) 흉막액 PPD 항체의 결핵성 흉막염 진단기준을 0.091로 할 경우 진단적 예민도는 55.0%, 94.7%를 나타내었다. 5) 흉막액 LAM-B 항체의 결핵성 흉막염 진단기준을 0.337로 할 경우 진단적 예민도는 50.0%, 특이도는 94.7%를 나타내었다. 5)결핵성 흉막염군의 진단 양성률은 PPD 피부반응 검사, 흉막액의 양, 활동성 폐결핵 동반여부 등에 의해 영향받지 않았다. 결론: PPD와 LAM-B에 대한 IgG 측정은 결핵성 흉막염의 진단에 도움을 줄 것으로 사료된다. 아울러 PPD와 LAM-B에 대한 IgG는 수동적으로 흉막조직을 통해 혈청에서 흉막액으로 이동함을 제시하고 있다.

• BMB Reports
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• 제30권5호
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• pp.326-331
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• 1997

To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

• 약학회지
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• 제40권4호
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• pp.422-427
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• 1996

The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-$d_2$, L-Phe-$d_5$, L-Trp-$d_5$, L-Tyr-2,6-$d_2$ and L-[1-$^{13}C$]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C$-$^{15}N$ double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the $V_H$ domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.

• Parasites, Hosts and Diseases
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• 제60권3호
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• pp.181-185
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• 2022

Strongyloides stercoralis infection is not endemic in the Republic of Korea (Korea) with a positivity rate of <1% in stool examination. However, there is a risk of hyperinfection in immunosuppressed individuals. It is necessary to determine the seropositivity of S. stercoralis antibodies in Korea. This study investigated the seropositivity of S. stercoralis antibodies in the southeastern area of Korea. From January 2019 to June 2021, serum samples were collected from participants who visited the study center in the southeastern region of Korea for routine health check-ups. We determined serum levels of specific anti-Strongyloides IgG antibodies in 834 samples by enzyme-linked immunosorbent assay. We observed that 92 samples (11.0%) tested showed a positive response. The age of the participants was 51±10.7 years, and 43.4% of them were men. The antibody positivity rate based on the location of the participants' residence were 12.3% (Gyoungsangnam-do), 10.2% (Busan), and 10.1% (Ulsan), respectively. Total eosinophil count was associated with positive test results (154.8±152.0 per mm³ versus 202.1±178.9 per mm³, P=0.006). Logistic regression analysis revealed that blood eosinophil count, age above 50 years, and residence in Sacheon were factors associated with the positive status of S. stercoralis antibody. Our finding suggests that it is necessary to test for S. stercoralis in actual clinical settings in Korea.

• Parasites, Hosts and Diseases
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• 제24권2호
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• pp.159-170
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• 1986

1984년 1월부터 1986년 6월까지 30개월간, 확진된 뇌 유구낭미충증 환자 69명에 대하여 혈청학적 추적검사를 실시하였다. 환자는 모두 치료전에 뇌전산화 단층 촬영을 실시하였고 효소면역측정법에 의하여 혈청 및 뇌척수액의 특이 IgG 항체가를 측정하였다. 프라지콴텔에 의한 약물 치료는 $3{\times}25mg/kg$의 용량으로 $4{\sim}14$일간 실시하였다. 추적기간 및 회수는 환자마다 달랐으며 최대 추적기간은 22개월이었다. 추적검사용 혈청 및 뇌척수액은 $-40^{\circ}C$에 보관한 후 낭액항원, 낭벽항원 및 두절항원에 대한 IgG 항체가를 일시에 측정하였다. 프라지콴텔 투여후 $1{\sim}4$개월 사이에 환자의 대부분에서 혈청 및 뇌척수내 특이 IgG 항체가는 일시적으로 상승하였다. 치료 직후부터 항체가가 저하하는 환자가 있었으나 소수이었다. 프라지콴델 투여시의 부작용을 줄이기 위해 덱사메타손을 투여하면 항체가 상승은 억제되었다. 프라지콴텔 치료후 혈청내 항체가 평균치의 변동이 뇌척수액내 항체가 평균치의 변동보다 심하였고, 항체가의 변화율은 낭벽항원이나 두절항원에 대한 항체에서 낭액항원에 대한 항체에서 보다 컸으나 항체가 변동폭은 낭액항원에 대한 항체가 더 컸다. 대상 환자 69명중 혈청학적으로 음성이었던 6명중 1명은 치료후 뇌척수액내 낭액항원에 대한 항체가가 양성범위로 전환하였다. 프라지콴텔 투여후 항체가의 상승은 뇌 유구낭미충증의 혈청학적 진단에서 보조적으로 이용할 수 있다고 생각하였으며 추적기간으로는 투여후 1개월이 적합하다고 생각하였다. 치료후 6개월내지 22개월까지 추적검사한 환자 15명중 혈청과 뇌척수액에서 모두 음성으로 전환한 예는 1례뿐이었고 혈청이나 뇌척수액 어느 하나에서라도 음성으로 전환한 예도 3례에 불과하였다. 만성 경과를 취한 심한 뇌 유구낭미충증 환자에서의 혈청학적 음성전환은 관찰 기간중 없었다. 추적 검사증 급성 뇌염증상이 나타난 만성환자에서는 일시적으로 두절항원에 대한 항체가가 상승하고 있었다. 이상의 결과에서 뇌 유구낭미충증 환자를 프란지콴텔로 치료한 다음 면역효소측정 법으로 혈청학적 추적검사를 실시할 경우 투약후 매 1년에 한번씩 실시하면 완치된 환자를 감별할 수 있을 것으로 생각하였다.

• Parasites, Hosts and Diseases
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• 제54권5호
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• pp.637-643
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• 2016

The immune correlate of host resistance induced by reinfection of Trichinella spiralis remains unclear. In this study, we investigated immune correlates between the resistance and serum IgG antibody level, $CD23^+$ $IgM^+$ B cells, and eosinophil responses induced by T. spiralis reinfection. Mice were primarily infected with 10 or 100 T. spiralis larvae (10 TS, 100 TS), respectively, and after 4 weeks, they were challenge infected with 100 T. spiralis larvae (10-100 TS, 100-100 TS). Upon challenge infections, 10-100 TS mice induced significantly higher levels of T. spiralis-specific total IgG antibody responses in sera and antibody secreting cell responses in spleens compared to 100-100 TS mice, resulting in significantly reduced worm burdens in 10-100 TS mice (60% and 70% reductions for adult and larvae, respectively). Higher levels of eosinophils were found in mice primarily infected with 10 TS compared to those of 100 TS at week 8 upon challenge. $CD23^+$ $IgM^+$ B cells were found to be increased significantly in mice primarily infected with 10 TS. These results indicate that primary infection of 10 larvae of T. spiralis, rather than 100 larvae, induces significant resistance against reinfection which closely correlated with T. spiralis-specific IgG, eosinophil, and $CD23^+$ $IgM^+$ B cell responses.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1194-1199
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• 1997

The effect of hot taste preference on selected immune responses was investigated in human peripheral immunocompetent cells. Human lymphocytes and natural killer(NK) cells were prepared at a concentration of 2$\times$$10^{6}$ cells/ml in RPMI-1640 containing 10% fetal bovine serum. Lymphocytes proliferation was determined with the [$^{3}H$]-thymidine pulse for 18hrs after concanavalin A, phytohemagglutinin, Salmonella typhimurium mitogen, or media alone. NK cell activity was measured by cytolysis of $^51Cr$-labeled target cells K562. Serum antibodies levels such as IgM, IgG, IgA were also measured by ELISA method. There was no difference of serum IgM level among the groups, but IgG and IgA levels were greater in the group with hot taste preference than those of the group without hot taste preference. In lymphocytes of the group with hot taste preference there was a greater mitogen-induced lymphocyte proliferative responses compared to the group without hot taste preference. In addition, NK cell activity in group with hot taste preference was lower than that of the group without hot taste preference. These results suggest that the eating habit of spicy food containing hot components may affect immune status by modulating selective immunocompetent cells function.