• Title/Summary/Keyword: serum IgG antibody

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Antigen analysis of Toxoplasma gondii Iysate and excretory-secretory materials by enzyme-linked immunoelectrotransfer blot (EITB) (효소면역 전기영동이적법에 의한 톡소포자충 용해물 및 분비 항원의 분석)

  • 안명희;손혁진
    • 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).

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Serological Investigation of the Infection Rate of Chlamydophila pneumonia among Residents of a Single University Dormitory (일개 대학 기숙사 거주 학생에 있어서 Chlamydophila pneumoniae의 혈청학적 감염률 조사)

  • Ryu, Jea Ki;Kim, Hyun-Kyung;Kim, Dong-Chan;Lee, Suk Jun
    • Journal of Life Science
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    • v.24 no.3
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    • pp.318-322
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    • 2014
  • Chlamydophila pneumonia is a common cause of community-acquired pneumonia throughout the world. It causes mild pneumonia or bronchitis in adolescents and young adults. Older adults may experience more severe disease and repeated infections. To the best of our knowledge, no study has attempted to investigate the prevalence of C. pneumonia in a closed community in Korea. We compared the infection rate of C. pneumonia among university dormitory residents using the miro-immunofluorescence (MIF) method. Antibody titers of IgG (1:32 or more) indicate past infection of C. pneumonia. A recent infection was defined as serum with a high titer of IgG (1:512 or more) or a positive IgM (1:16 or more). The past infection rate of C. pneumonia among the university dormitory residents was 71.7%. The recent infection rate of C. pneumonia according to IgG and IgM titers was 28.3% and 23.3%, respectively. The past infection positive rate according to the number of residence months was 1 month (50%), 7 months (71.4%), 13 months (66.7%), and 35 months (89.5%). The recent infection positive rate according to IgG antibody titers was 1 month (50%), 7 months (28.6%), 13 months (33.3%), and 35 months (10.5%). The recent infection rate of C. pneumonia according to IgM antibody titers was 1 month (41.7%), 7 months (28.6%), 13 months (26.7%), and 35 months (5.3%). The results suggest that the past infection rate of C. pneumonia is increased by the number of residence months in a closed community and that the recent infection rate of C. pneumonia according to IgG and IgM serological tests is decreased by the number of residence months.

Immunogic Study on the Effect of Gamidaegang-tang on Collagen II - Induced Arthritis in Mice (가미대강활탕(加味大羌活湯)이 Collagen II로 유발된 관절염 생쥐모델에 미치는 면역학적 연구)

  • Kim, Tae-Young;Oh, Min-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1095-1105
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    • 2009
  • This study was carried out to know the immunological effect of GDGT on CIA(collagen induced arthritis) mice, a model of rheumatoid arthritis. For this purpose, GDGT was orally administerd to mice with arthritis induced by collagen II and then value of cytotoxicity on hFLSs and liver, the arthritis index, immunocyte in paw joint and DNL, rheumatoid factor (IgG and IgM), collagen II specific antibody in the serum were measured. The cytotoxicity were not shown on hFLSs and liver. The arthritis index decreased significantly after 3 week. In total cell counts of DLN and paw joint, there was a significant increase in DLN and significant decrease in paw joint. In DNL, $CD19^+$, $CD3^+$, $CD4^+$, $CD3^+/CD69^+$, $CD8^+$, $CD4^+/CD25^+$, $ CD3^+/CD49b^+$ cells increased significantly. In Paw joints, $CD3^+$, $ CD4^+$, $CD4^+/CD25^+$ cells decreased significantly. The level of serum IgG and IgM decreased significantly. The level of collagen II in the serum was decreased significantly. Marginal erosion, necrotic chodrocytes, cartilage and bone degradation were improved in histological section of paw joints. The results present significant immunological effect of GDGT on rats with arthritis induced by collgen II. So we expect that GDGT should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.

Effect of the Crude Polysaccharides Fraction from Eleutherococcus senticosus as a Immunoadjuvant to Soluble Antigens (BSA and OVA) (오가피 조다당의 단백질 항원 (BSA and OVA)에 대한 면역증강효과)

  • 황수현;하은숙;유광원;신광순;이상훈;이재권;이경호;윤택준;박우문
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.167-175
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    • 2003
  • Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.

Effect of Ecklonia cava Water Extracts on Inhibition of IgE in Food Allergy Mouse Model (식품 알레르기 마우스 모델에서 감태 물 추출물의 IgE 분비 억제 효과)

  • Song, Eu-Jin;Lee, Chung-Jo;Kim, Koth-Bong-Woo-Ri;Jung, Ji-Yeon;Kwak, Ji-Hee;Choi, Moon-Kyoung;Kim, Min-Ji;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1776-1782
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    • 2010
  • This research was done to verify the effect of Ecklonia cava water extracts (ECWE) on inhibition of allergic reactions using ovalbumin (OVA)-immunized food allergy mouse model. For in vitro test, $10\sim100{\mu}g$/mL of ECWE and OVA were added to splenocytes obtained from OVA-immunized mice. The significant reduction of IgE antibody level in culture supernatants of splenocytes was shown in ECWE adding group at all tested concentrations. In addition, ECWE decreased IL-4 and IFN-$\gamma$ levels in supernatants of splenocytes. To confirm the effect of ECWE in in vivo test, ECWE was injected to peritoneal cavity of OVA-immunized mice. Subsequently, IgE level was measured in serum and cultured supernatants of splenocytes. As a result, the injection of ECWE (5 and 10 mg/kg BW) significantly attenuated the secretion of IgE antibody in both serum and splenocytes. In conclusion, the present study indicates that ECWE could suppress in a food allergy mouse model through the inhibition of IgE secretion.

Enhanced Antigen Delivery Systems Using Biodegradable PLGA Microspheres for Single Step Immunization

  • Cho, Seong-Wan;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.443-450
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    • 2006
  • To demonstrate their possibilities as an enhanced vaccine delivery system, protein-loaded Poly lactide glycolide copolymer (PLGA) microspheres were prepared with different physical characteristics. Ethyl acetate (EA) solvent extraction process was employed to prepare microspheres and the effects of process parameters on drug release properties were evaluated. The biodeuadability of microspheres was also evaluated by the pH change and GPC (Gel permeation chromatography). Primary IgG antibody responses in BALB/c mice were compared with protein saline solutions as negative controls and adsorbed alum suspensions as positive controls after single subcutaneous injection for in vivo studies. The microspheres showed a erosion with a highly porous structure and did not keep their spherical shape at 45 days and this result could be confirmed by GPC. In vitro release of proteinous drug showed initial burst effect in all batches of microspheres, followed by gradual release over the next 4 weeks. PLGA microspheres were degraded until 45 days and the secondary structure of OVA was not affected by the preparation method. Enzyme-linked immunosorbent assays demonstrated that the single subcutaneous administrations of OVA-loaded PLGA microspheres induced enhanced serum IgG antibody response in comparison to negative and positive controls. These results demonstrated that microspheres providing the controlled release of antigens might be useful in advanced vaccine formulations for the parenteral carrier system.

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A Case of Autoimmune Chronic Pancreatitis in a Child (자가면역성 만성 췌장염으로 진단된 소아 1예)

  • Choi, In-Young;Jin, So-Hee;Choi, Kyung-Dan;Kim, Kyung-Mo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.10 no.2
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    • pp.215-220
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    • 2007
  • We present a case of autoimmune chronic pancreatitis in a previously healthy child without any history of autoimmune disease. A 12-year-old boy was admitted to the hospital with abdominal pain. The serum amylase, lipase, and IgG levels were elevated and autoantibodies (antinuclear antibody, antineutrophil antibody) were detected. An abdominal CT (computed tomographic) scan revealed diffuse enlargement of the pancreas. ERCP (endoscopic retrograde cholangiopancreaticography) demonstrated an irregular stricture of the main pancreatic duct in the pancreas tail. After two years of oral steroid and immunosuppressive drug therapy, the clinical, laboratory and radiological findings were improved. The patient has been symptom-free for 18 months after the discontinuation of medication.

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Serologic Response to Pneumocystis National University Hospital Patients

  • Sung-Tae Hong
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.355-362
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    • 1991
  • A total of 2,580 sera of the patients who were consulted to the serology laboratory of the Seoul National University Hospital were collected in 1990. The sera were screened by micro-ELISA to detect IgG antibody reacting with Pneumocystis carinii antigen. The absorbances were 0.00 to 1.41 and mean 0.27±0.253. As the positive criterion was set absorbance 0.2 or more with 70% sensitivity, total positive rate was 44.4oA. Mean absorbances and positive rates were higher in children than in adults; 0.40 and 62.9% in 0 year group, 0.50 and 81.2% in 1 year group, 0.41 and 66.0% in 2-3 year group, 0.33 and 61.4% in 4-5 year group, 0.25 and 42.3% in 6-10 year group respectively. In the age groups over 11 years, the absorbances were in range of 0.16 to 0.23 and the positive rates were 26.1% to 41.5%. The present level of absorbances and positive rates could be regarded similar with those in normal Korean population. The present findings suggest that most humans are exposed to Pneumocystis within 2 years after birth and meet much less new antigenic challenge after 11 years in Korea.

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$HgCl_2$ Dysregulates the Immune Response of Balb/c Mice (수은에 의한 마우스의 면역반응 조절장애)

  • Ki, No-Suk;Koh, Dai-Ha;Kim, Chong-Suh;Lee, Jung-Sang;Kim, Nam-Song;Lee, Hwang-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.1 s.45
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    • pp.11-24
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    • 1994
  • The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

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Serum Antigen and Antibody Detection in Echinococcosis: Application in Serodiagnosis of Human Hydatidosis

  • Sadjjadi, Seyed Mahmoud;Sedaghat, Farzaneh;Hosseini, Seyed Vahid;Sarkari, Bahador
    • Parasites, Hosts and Diseases
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    • v.47 no.2
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    • pp.153-157
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    • 2009
  • Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.