• Parasites, Hosts and Diseases
• /
• v.29 no.4
• /
• pp.339-354
• /
• 1991

Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

• Food Science of Animal Resources
• /
• v.27 no.4
• /
• pp.409-415
• /
• 2007

This experiment was conducted to investigate the effects of dietary enzyme mixture fortified with ${\beta}-glucanase$ on the growth performance, serum components and meat quality of broiler chicks. 31,800 Ross 208 male broiler chicks were randomly allotted into 2 groups, the control and 0.3% enzyme diet with ${\beta}-glucanase$ supplementation groups. Control group chicks were fed the control (corn-soybean meal based) diet and the treatment group chicks were fed the 0.3% enzyme mixture supplemented with ${\beta}-glucanase$. The growth performance, serum components and meat qualities such as pH, color, water holding capacity, cooking loss, and shearing force of meats were investigated. The results showed that the growth performance of chicks fed the 0.3% enzyme mixture diet were improved compared to that of the control group, as much as 5% in growth rate, 19% in average weight, 6.8% in performance index, and 5.5% in feed efficiency. Although, there were no significant differences in the muscle color degrees ($L^*a^*b^*$) and shearing force between the control group and experimental group, the water holding capacity and cooking loss of the experimental group were significantly higher than those of control group (p<0.05). The antibody titers in serum against the antigens of Newcastle disease and Infectious Bursal disease were higher in the experimental group than in the control group. Altogether, these suggest that the broiler diet containing 0.3% enzyme mixture fortified with ${\beta}-glucanase$ activity can improve the growth performance, immune reaction, and meat quality of broiler chicks.

• Laboratory Medicine and Quality Assurance
• /
• v.40 no.2
• /
• pp.51-69
• /
• 2018

As part of the immunoserology program of the Korean Association of External Quality Assessment Service, we organized two trials on the external quality assessment of hepatitis viral markers in 2016 and 2017. The hepatitis viral antigens and antibodies program consisted of 10 test items. We delivered two and three types of pooled sera specimens to 965 and 965 institutions for the first and second trials of external proficiency testing in 2016, respectively. The number of participating laboratories was 915 (94.8%) and 913 (95.0%) in the first and second trials in 2016, respectively. We also delivered three kinds of pooled sera specimens to 936 and 1,015 institutions for the first and second trials of external proficiency testing in 2017, respectively. The number of participating laboratories was 920 (98.3%) and 996 (98.1%) in the first and second trials in 2017, respectively. The most commonly tested items were hepatitis B surface antigen, followed by the antibodies to hepatitis B surface antigen, anti-hepatitis C virus, hepatitis B envelope antigen, antibodies to hepatitis B envelope antigen, anti-hepatitis A virus and antibodies to hepatitis B core antigen. The most frequently used methods for detecting viral markers were the chemiluminescence immunoassay and the electrochemiluminescence immunoassay, but they yielded a few-false positive results due to the matrix effect. The immunochromatographic assay yielded false-negative results for anti-hepatitis A virus due to low sensitivity. Continuous improvement in the quality of viral hepatitis testing through participation in the survey seems necessary.

• The Korean Journal of Blood Transfusion
• /
• v.23 no.2
• /
• pp.115-126
• /
• 2012

Background: Umbilical cord (UC) is a promising source of mesenchymal stromal cells (MSCs). We compared the characteristics of MSCs from cryopreserved UC with those from fresh tissues, and demonstrated the possibility of UC cryopreservation for acquisition of MSCs from cryopreserved UC. Methods: Each UC was sliced into two types ($1{\sim}2mm^3$ vs. 0.5 cm), and cryopreserved in liquid nitrogen using different media (autologous cord blood plasma, aCBP vs. RPMI 1640). A fresh aliquot of $1{\sim}2mm^3$-sized UC was used as control tissue. After one week, the cryopreserved tissues were thawed and cultured. For the 0.5 cm UC, a slicing step into $1{\sim}2mm^3$ was needed. Cell count, viability, proliferative activity, and surface antigens were determined from harvested MSCs. Several growth factors (EGF, IGF-1, PDGF, TGF-${\beta}$, bFGF, and VEGF), were measured from the culture supernatant. Results: Eleven UC were enrolled in the study. Efficiencies of obtaining MSCs were higher in cryopreserved UC using RPMI 1640, compared with use of aCBP; the same result was observed for 0.5 cm sized UC, compared with $1{\sim}2mm^3$ sized UC. No difference in proliferative activity was observed between MSCs from fresh and cryopreserved UC. The amount of growth factors in culture supernatant using RPMI 1640 was larger than that of fresh tissues. Conclusion: We obtained growth factors from the supernatant as well as MSCs from cryopreserved UC. As with a cord blood bank, in the future, cryopreservation of UC for acquisition of both MSCs and growth factors would be possible in a time of need.

• Journal of Life Science
• /
• v.28 no.12
• /
• pp.1416-1423
• /
• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• Journal of Applied Biological Chemistry
• /
• v.66
• /
• pp.447-454
• /
• 2023

In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.14 no.2
• /
• pp.203-207
• /
• 2010

Purpose: Hepatitis B is infection caused by Hepatitis B virus (HBV). Currently, there are several methods, Kits and equipments for conducting Hepatitis B test. Due to ununiformed methods, it would cause some differences. To manage these differences, it needs process evaluating function of test system and reagent using particular standard substance. The aim of this study is to investigate tendency of RIA method's reagent used in Asan Medical Center through comparing several other test reagents using national standard substance. Materials and Methods: The standard substance in National Institute of Food and Drug Safety Evaluation's biology medicine consists of 5 things, 4 antigens and 1 antibody. We tested reagents using A, B company's Kits according to each test method. All tests are measured repeatedly to obtain accurate results. Results: Test result of "HBs Ag Mixed titer Performance panel" is obtained match rate compared S/CO unit standard with RIA method and EIA 3 reagents, CIA 2 reagents is that company A's reagent is 94.4% (17/18), 83.3% (15/18), B is 88.9% (16/18), 77.8% (14/18). Test result of "HBs Ag Low titer Performance panel" is obtain that EIA 2 reagents is shown 7 posive results, CIA 3 reagents is 11, and RIA method's company A's reagent is 3, B is 2 of 13 in low panel. "HBV surface antigen 86.76 IU/vial" tested dilution. A is obtain positive results to 600 times(0.14 IU/mL), B is 300 times (0.29 IU/mL). Case of "HBV human immunoglobulin 95.45 IU/vial", A is shown positive result to 10,000 times (9.5 mIU/mL) and B is 4,000 times (24 mIU/mL). Test result of "HBs Ag Working Standards 0.02~11.52 IU/mL" is shown that Company A's kit concentration level was 0.38IU/mL, company B was 2.23 IU/mL and higher level of concentration was positive results. Conclusion: When comparing various test reagents and RIA method according to National Standard substances for Hepatitis B test, we recognized that there were no significant trends between reagents. For hepatitis B virus antigen-antibody titers even in parts of the test up to 600 times the antigen, antibodies to 10,000 times the maximum positive results could be obtained. Therefore, we confirmed that results from Asan Medical Center are performed smoothly by reagents and system for hepatitis B virus test.

• Journal of Animal Science and Technology
• /
• v.48 no.6
• /
• pp.933-942
• /
• 2006

This experiment was carried out to investigate effects of honeybee venom treatment on the humoral immune response in pigs. Corresponding author : S. K. Cho, Dept. of Animal Sci. Chung-Buk National University, Kaesin-dong, Cheongju, 361-763, Korea. phone : 043-261-2551. E-mail : deercho@chungbuk.ac.kr To investigate effects of natural honeybee venom on the concentration of immunoglobulin G, A, and M, 20 piglets(LY×D) from 3 sows were allocated into two groups bee venom-treated group(10 piglets) and non-treated control(10 piglets). Natural honeybee venom was treated at 0, 3, 6 days after birth and the acupoints were Hai-men(ST-25), Du-kou(CV-8) and Jiao-chao(GV-1) points at 0, 3 days after birth and the regions of castration and tail amputation point at 6 days. Control group was injected 1㎖ of saline to the same site. Concentrations of IgG, A, and M were measured with immunoturbidimetric method at 0, 3, 7, 14, and 21 days after treatment. To investigate the effect of bee venom on the production of antibodies against hog cholera and atrophic rhinitis vaccines that were used as indicator antigens, 40 piglets(LYxD) from 5 sows were grouped as bee venom-treated group (20 piglets) and control group(20 piglets). Natural honeybee venom was treated at 0, 3days(castration, tail amputation) and 21days after birth. The acupoints were Hai-men(ST-25), Du-kou(CV-8) and Jiao-chao (GV-1) points at 0 day, the regions of castration and tail ampution at 3 days and Jiao-chao(GV-1) and Bai-hui(GV-20) points at 21days after birth(weaning). Control group was injected 1ml of saline to the same site. Atrophic rhinitis vaccine was injected twice at 24 and 44 days after birth and hog cholera vaccine was also injected twice at 44 and 64 days after birth. Antibody titers against Bordetella bronchiseptica and hog cholera virus were measured by using tube agglutination and ELISA tests at 24, 34, 44, 54 and 74 days after birth. Concentrations of IgG of treated group were 339.52, 366.48, 296.52, 242.06 and 219.06mg/dl at 0, 3, 7, 14 and 21 days after birth, respectively. In contrast, concentrations of IgG in control group were respectively 347.10, 334.14, 243.28, 205.18 and 191.58mg/dl during same periods with treated group. Concentrations of IgG at 0 day was not significantly different between the treated group and control group but treated group were significantly increased by 10.28% at 3 days after birth (P<0.02), 21.88% at 7 days after birth(P<0.01), 18.0% at 14 days after birth(P<0.07) and 14.3% at 21 days after birth(P<0.01). Concentrations of IgA and Ig M were not significantly different. Antibody titers against hog cholera virus were significantly increased by 57.0% at 24 days after birth(P<0.03), 74.6% at 34 days after birth (P<0.006), 48.6% at 44 days after birth(P<0.017), 45.0% at 54 days after birth(P<0.16) and 44.4% at 74 days after birth (P<0.006) in bee venom treated group in comparison with control group. Antibody titers against the Bordetella bronchiseptica was significantly increased in Beevenom treated group as 9.1% (P<0.32) at 24days, 39.7% (P<0.002) at 34days, 31.9% (P<0.02) at 44days, 33.4% (P<0.01) at 54days and 57.3% (P<0.007) at 74 days after birth when compared with those of control group pigs. Collecting together, the results in this study showed that immune responses were increased by treatment of natural honeybee venom to pigs. These results suggested that the treatment of bee venom could be used effectively for the increase of productivity in livestock industry.

• Journal of Food Hygiene and Safety
• /
• v.20 no.2
• /
• pp.118-122
• /
• 2005

Immu-Forte (Dong-Ahm Bio's. Corp., Korea) was evaluated fir its effectiveness as a nonspecific immunostimulator in mice. The effects of Immu-Forte were determined by analysis of cytokines using ELISh and phenotype of leukocyte subpopulations using monoclonal antibodies specific to mouse leukocyte differentiation antigens and flow cytometry. CD4 T cells, CD8 T cells, macrophages, IL-12 and IFN-r in Immu-Forte EX-treated middle dose group increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. All T cells, all B cells, macrophages, IL-2, IL-4 and IL-12 in Immu-Forte EX-treated low dose uoup increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte soy-treated group, CD4 T cells, IL-2, IL-4 and IL-12 were significantly higher in high dose-treated group, and CD 4 T cell, macrophages, IL-2, IL-4 and IL-12 were significantly higher in middle dose-treated group, and all T cell, IL-2, IL-4 and IL-12 were significantly higher in low dose-treated group. In the Itnmu-Forte A-treated group, macrophages, m cells and IL-12 in high dose-treated group and all T cells, macrophages, NK cells, IL-2, IL-4 and IL-12 in middle dose-treated group and NK cells in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte F-treated Group, all B cells, IL-4 and IL-12 in high dose-treated group and all T cells, aBl B cells, CD 4 T cells, CD8 T cells, macrophage, IL-2, IL-4, IL-12 and IFN-r in middle dose-treated group and NK cells and IL-12 in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In conclusion, the study has demonstrated that Immu-Forte had an immunostimulatory effect on mice through proliferation and activation of mouse immune cells.

• Tuberculosis and Respiratory Diseases
• /
• v.53 no.3
• /
• pp.294-308
• /
• 2002

Background : The cell-mediated immune response plays an important role in tuberculosis. After being activated by mycobacterial antigens, T lymphocytes express a high affinity receptor (IL-2R) for interleukin-2 (IL-2) on their own surface and release a soluble fraction of the IL-2 receptor (sIL-2R) from the cell membrane into the circulation. Neopterin is a metabolite of guanosine-triphosphate, which is produced by stimulated macrophages under the influence of IFN-$\gamma$ with a T lymphocyte origin. Therefore, the utility of sIL-2R, IFN-$\gamma$ and the neopterin levels as immunologic indices of the cell-mediated immune response and severity of disease in patients with pulmonary tuberculosis was assessed. Methods : The serum sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 39 patients with pulmonary tuberculosis, 6 patients with tuberculous lymphadenitis prior to treatment and 10 healthy subjects. The serum and pleural sIL-2R, neopterin and ADA levels were measured in 22 patients with tuberculous pleurisy. The patients with pulmonary tuberculosis were divided into a mild, moderate and severe group according to the severity by ATS guidelines. To compare the results from these patients with those of the pretreatment levels, the sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 36 of the 39 patients(1 patient, expired; 2 patients were referred to a sanitarium) with pulmonary tuberculosis after 2 months of treatment. Results : 1) the serum sIL-2R and IFN-$\gamma$ levels were elevated in patients with tuberculosis when compared to those of healthy subjects (p>0.05). The neopterin concentration in the serum was significantly lower in patients with pulmonary tuberculosis($2967{\pm}2132.8$ pg/ml) than in healthy controls($4949{\pm}1242.1$ pg/ml)(p<0.05). 2) In the pulmonary tuberculosis group, the serum sIL-2R and IFN-$\gamma$ levels were higher in patients with severe disease than those in patients with mild and moderate disease. However, the neopterin levels declined as the pulmonary tuberculosis became more severe (p<0.01). 3) The mean serum sIL-2R and IFN-$\gamma$ levels declined from $1071{\pm}1139.4$ U/ml to $1023{\pm}1920.9$ U/ml(p>0.05), $41{\pm}52.8$ pg/ml to $22{\pm}23.9$ gm/ml(p<0.05), respectively, after 2 month of treatment. The mean serum neopterin levels increased from $3158{\pm}2272.6$ pg/ml to $3737{\pm}2307.5$ pg/ml(p>0.05) after a 2 month of treatment. These findings were remarkable in the severe group of pulmonary tuberculosis with a clinical correlation. 4) In the patients with tuberculous pleurisy, the serum sIL-2R and ADA were significantly higher than those in the pleural fluid, However, the neopterin levels in the sera and pleural effusion were similar. Conclusion : On the basis of this study, sIL-2R, IFN-$\gamma$ and neopterin measurements may not only provide an insight into the present state of the cell-mediated immune response, but also serve as parameters monitoring of the prognosis of the disease, particularly in patients with severe pulmonary tuberculosis. In addition, an assay of the pleural sIL-2R levels might signal a stimulated local immunity including T cell activation in the tuberculous pleural effusion.