• IMMUNE NETWORK
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• v.11 no.2
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• pp.100-106
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• 2011

Background: Disparities of Minor H antigens can induce graft rejection after MHC-matched transplantation. H60 has been characterized as a dominant antigen expressed on hematopoietic cells and considered to be an ideal model antigen for study on graft-versus-leukemia effect. Methods: Splenocytes from C57BL/6 mice immunized with H60 congenic splenocytes were used for establishment of H60-specific CTL clones. Then the clones were characterized for proliferation capacity and cytotoxicity after stimulation with H60. Clone #14, #15, and #23 were tested for the TCR binding avidity to H60-peptide/$H-2K^b$ and analyzed for TCR sequences. Results: H60-specific CTL clones showed different levels of proliferation capacity and cytotoxic activity to H60-stimulation. Clones #14, #15, and #23 showed high proliferation activity, high cytotoxicity, and low activities on both aspects, respectively, and have TCRs with different binding avidities to H60-peptide/$H-2K^b$ with $t_{1/2}$ values of 4.87, 6.92, and 13.03 minutes, respectively. The TCR usages were $V{\alpha}12D-3-01+J{\alpha}11-01$ and $V{\beta}12-1-01+D{\beta}1-01+J2-7-01$ for clone #14, $V{\alpha}13D-1-02+J{\alpha}34-02$ and $V{\beta}13-1-02+D{\beta}2-01+J{\beta}2-7-01$ for clone #15, and $V{\alpha}16D+J{\alpha}45-01$ and $V{\beta}12-1-01+D{\beta}1-01+J{\beta}2-5-01$ for clone #23. Conclusion: The results will be useful for modeling GVL and generation TCR transgenic mouse.

• Journal of Oral Medicine and Pain
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• v.30 no.1
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• pp.15-23
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• 2005

Mucin glycoproteins are the primary carriers of the oligosaccharide moieties that constitute the blood group substances in human saliva. The aim of this study was to determine whether or not the conversion of either the A or B blood group antigens to the H antigen can occur during the degradation process of stored saliva samples. Forty subjects (20 subjects in each A and B blood group) identified as secretors were enrolled in this study. Fresh whole saliva samples and their clarified supernatants were stored at room temperature for 1 week. The conversion of the blood group antigens was detected by SDS-PAGE and immunoblotting. Among the subjects showing the conversion in whole saliva, glandular saliva samples were obtained from 8 subjects (4 subjects in each A and B blood group). Submandibular-sublingual saliva (SMSL) and a mixture of SMSL and parotid saliva (PS) were stored at room temperature for 1 week. The conversion of the blood group antigens was detected by the same method. The obtained results were as follows: 1. In the clarified samples of whole saliva, the A antigen was detected as being either intact (5%) or degraded molecules (95%) after the 1 week period. Conversion of the A antigen to the H antigen was detected in 5 subjects (25%). In the unclarified samples, the A antigen was either detected as degraded molecules (90%) or was not detected (10%). Conversion of the antigen had occurred in 4 subjects (20%). 2. In the clarified samples of whole saliva, the B antigen was detected as intact (20%) or as degraded molecules (65%) or was not detected (15%) after the 1 week period. Conversion of the B antigen to the H antigen was detected in 7 subjects (35%). In the unclarified samples, the B antigen was detected as intact (5%) or as degraded molecules (65%), or was not detected (30%). Conversion of the antigen was observed in 2 subjects (10%). 3. In the glandular saliva samples, only one of the four subjects displayed an antigenic conversion from the A to H antigen or from the B to H antigen. The conversion had occurred in both the SMSL samples and the SMSL and PS mixture. No degradation of the antigens was detected in the other three samples of the A or B blood groups, nor was there any conversion. The results demonstrated that conversion of the blood group antigens could occur in saliva, and suggested that the enzymes responsible for the conversion are present in saliva. Further studies on the origin and activity of the specific glycosidases in saliva as well as quantitative measurements of the antigenic conversion will be needed.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.338-343
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• 2003

Various factors, such as the adsorption pH, adjuvant dose, and adjuvant age, which affect the adsorption degree and immunogenicity of an antigen, were investigated. In addition, the effect of pH, antigen content, and adjuvant content on immunogenicity was also studied through animal experiments. Within the ranges studied, a low pH for adsorption, freshly preformed gel, and low pH formulation for the combined DTwP-HepB vaccine were preferrable for the adsorption of the antigens. In addition, a higher DT content was found to have a positive effect on the HBsAg immunogenicity in the combined vaccine. Accordingly, considering the factors affecting the adsorption rate and immunogenicity of the antigens, a novel DTwP-HepB vaccine (40 Lf/ml of diphtheria toxoid, 15 Lf/ml of tetanus toxoid, 20 OU/ml of detoxified whole cell pertussis, $24\;\mu\textrm{g}$ of HBsAg, $24\;\mu\textrm{g}\;Al/ml\;of \;Al(OH)_3\;gel,\;776\;\mu\textrm{g}\; Al/ml\;of\;AIPO_4\;gel$, and pH 7.1) was developed, whose immunogenicity was comparable to the case of administrating, separately and simultaneously, a combined DTwP vaccine (40 Lf/ml of diphtheria toxoid, 15 Lf/ml of tetanus toxoid, 20 OU/ml of detoxified whole cell pertussis, $300\;\mu\textrm{g}\;Al/ml\;of\; AIPO_4\;gel$, and pH 7.1) and mono HepB vaccine [$Hepavax^{\circledR},\;24\;\mu\textrm{g}/ml$ of HBsAg and $500\;\mu\textrm{g}\;Al/ml\;of\;Al(OH)_3\;gel$], which satisfies the potency criteria of the K-FDA for a combined DTwP vaccine and mono HepB vaccine.

• Biomolecules & Therapeutics
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• v.19 no.1
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• pp.77-83
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• 2011

Sulforaphane is an isothiocyanate found in cruciferous vegetables that has been reported to be an effective cancer preventive agent inducing growth arrest and/or cell death in cancer cells of various organs. This paper reports that sulforaphane exerts immunomodulatory activity on the MHC-restricted antigen presenting function. Sulforaphane efficiently increased the class II-restricted presentation of an exogenous antigen, ovalbumin (OVA), in both dendritic cells (DCs) and peritoneal macrophages in vitro. The class II-restricted OVA presentation-enhancing activity of sulforaphane was also confirmed using mice that had been injected with sulforaphane followed by soluble OVA. On the other hand, sulforaphane did not affect the class I-restricted presentation of exogenous OVA at concentrations that increase the class II-restricted antigen presentation. At a high concentration ($20\;{\mu}M$), sulforaphane inhibited the class I-restricted presentation of exogenous OVA. Sulforaphane did not affect the phagocytic activity of the DCs, and the cell surface expression of total H-$2K^b$, B7-1, B7-2 and CD54 molecules, even though it increased the expression of I-$A^b$ molecules to a barely discernable level. These results show that sulforaphane increases the class II-restricted antigen presenting function preferentially, and might provide a novel insight into the mechanisms of the anti-cancer effects of sulforaphane.

• Pediatric Infection and Vaccine
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• v.14 no.1
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• pp.97-103
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• 2007

Purpose : Lewis antigen has been known to have a role in the attachment of H. pylori to the gastric mucosa, but its expression pattern in children with H. pylori infection is still unclear. The recently described blood group antigen-binding adhesin BabA is known to mediate adherence of H. pylori to Lewis B receptors on gastric epithelium. We investigated the expression of Lewis antigen in gastric mucosa of Korean children with H. pylori infection. Methods : The expression of Lewis A ($Le^a$), B ($Le^b$), X ($Le^x$), and Y ($Le^y$) was evaluated by immunohistochemistry in H. pylori positive biopsy specimens from 35 children (antral gastritis in 30, peptic ulcer in 5) and in H. pylori negative specimens from 19 children. PCR assays for cagA and babA2 gene of H. pylori were performed. Results : We confirmed the expression of $Le^a$ in 60%, $Le^b$ in 97%, $Le^x$ in 100%, and $Le^y$ in 100% of the superficial epithelium of the 35 H. pylori positive children. In H. pylori negative patients, $Le^a$, $Le^b$, $Le^x$, and $Le^y$ expression was 52%, 100%, 89%, and 100%, respectively. The cagA gene was detected in 65% and babA2 gene in 25% of 35 patients. No differences in neutrophil activity and chronic inflammation were found according to the presence of cagA and babA2 genes in H. pylori. Conclusion : $Le^b$, $Le^x$ and $Le^y$ antigen were highly expressed in gastric mucosa of Korean children, but they were not associated with the status of H. pylori infection and the positivity of babA2 gene. Further studies for other mucosal receptors and toxins are needed to define the immune responses to H. pylori infection in gastric mucosa of Korean children.

• Journal of Sericultural and Entomological Science
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• v.15 no.2
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• pp.1-7
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• 1973

It was found that the diapause in the silkworm egg is induced by the action of the diapause hormone secreted from the suboesophageal ganglion, and "esterase A" affects protein metabolism in oocyte and egg. In this connection, some changes in protein metabolism of silkworm egg according to embryonic developments could give some information on the diapause, using Ouchterlony Test. Antigenicity of the protein of silkworm egg was detected through antigen-antibody interaction among the extracts of rabbit blood. Furthermore, existence of the specific antigen was also detected according to embryonic development, using the adsorption test. The results were obtained as follows: 1 Detection of antigenicity The antigenicity of silkworm egg was ascertained by inoculating it into a rabbit, but positive results were shown in most of the silkworm eggs tested, whereas the antibody specific to a certain antigen was not detected. 2. Detection of the common antigen It was demonstrated that most of the antigen could incite the common antibodies, but the specific antibody formation was not detected in a few antigens, even though the nonspecific antibody formation was displayed. 3. Detection of the specific antigen It is suggested that there are the specific antigens detectable in each treated eggs by the adsorption test.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.374-378
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• 2004

The effects of various environmental factors such as pH (5, 6, 7, 8 and 9), temperature (30, 37 and $40^{\circ}C$) and rotational speed (150, 200 and 250 rpm) on the growth and the hepatitis B core antigen (HBcAg) production of Escherichia coli W3110IQ were examined in the present Study. The highest growth rate is achieved at pH 7, $37^{\circ}C$ and at a rotational Speed of 250 rpm which is 0.927 $h^{-1}$. The effect of pH on cell growth is more substantial compared to other parameters; it recorded a $123\%$ different between the highest growth rate (0.927 $h^{-1}$) at pH 7 and lowest growth at pH 5. The highest protein yield is achieved at pH 9, rotational speed of 250 rpm and $40^{\circ}C$. The yield of protein at pH 7 is $154\%$ higher compared to the lowest yield achieved at pH 5. There is about $28\%$ different of the protein yield for the E. coli cultivated at 250 rpm compared to that at 150 rpm which has the lowest HBcAg yield. The yield of protein at $40^{\circ}C$ is $38\%$ higher compared to the lowest yield achieved at $30^{\circ}C$.

• Bulletin of the Korean Chemical Society
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• v.18 no.1
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• pp.44-50
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• 1997

A method of dual-signal generation from two different enzymes was developed and utilized to simultaneously perform dual immunoassays in a single microwell. Two enzymes selected as tracers were horseradish peroxidase (HRP) and β-galactosidase (GAL). 3, 3', 5, 5'-Tetramethylbenzidine (TMB) and chlorophenolred-β-galactopyranoside (CPRG) as chromogenic substrates for the respective enzyme were used. Although the two enzymes showed their maximum activities at distinct pH conditions (pH 5.1 for HRP and 7.5 for GAL), the enzyme reactions were able to be concurrently carried out at pH 5.75 in a dual-substrate solution without signal loss. This performance was achieved by increasing TMB concentration two-fold, introducing potassium salt as activator of GAL reaction, and extending total reaction time 50%. The signal generation method was then used for dual-enzyme immunoassays to detect antibodies with co-immobilized Hepatitis C virus antigens (core and NS5) and a Hepatitis B virus antigen (PreS(2)) in a microwell. Dose-response curves of the assays revealed cooperativity between different antigen-antibody complex formation, which suggested that dual immunoassays can only be used for qualitative screening tests unless the antigens immobilized were spatially separated.

• Parasites, Hosts and Diseases
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• v.23 no.1
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• pp.95-101
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• 1985

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

• IMMUNE NETWORK
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• v.5 no.3
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• pp.163-171
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• 2005

Background: To perform the successful dendritic cell-based cancer immunotherapy one of the main issues to be solved is the source of antigen for DC pulsing. Limitations occur by using auto-tumor lysate due to the difficulties obtaining enough tumor tissue(s) quantitatively as well as qualitatively. In this study the possibility of allogeneic tumor cell lysate as a DC pulsing antigen has been tested in mouse melanoma pulmonary me tastasis model. Methods: B16F10 melanoma cells $(1{\timeS}10^5/mouse)$ were inoculated intra venously into the C57BL/6 mouse. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 (1,000 U/ml each) for 7 days and pulsed with lysate of either autologous B16F10 (B-DC), allogeneic K1735 (C3H/He origin; K-DC) or CloneM3 (DBA2 origin; C-DC) melanoma cells for 18 hrs. Pulsed-DCs $(1{\times}10^6/mouse)_{[CGP1]}$ were injected i.p. twice with one week interval starting from the day 1 after tumor cell inoculation. Results: Without observable toxicity, allogeneic tumor cell lysate pulsed-DC induced the significantly better anti-tumor response (tumor scale: $2.7{\pm}0.3,\;0.7{\pm}0.3\;and\;0.3{\pm}0.2$ for saline, B-DC and C-DC treated group, respectively). Along with increased tumor specific lymphocyte proliferations, induction of IFN-${\gamma}$ secretion against both auto- and allo-tumor cell lysates was observed from the DC treated mice. (w/B16F10-lysate: $44.97{\pm}10.31,\;1787.94{\pm}131.18,\;1257.15{\pm}48.27$, w/CloneM3 lysate: 0, $1591.13{\pm}1.83,\;1460.47{\pm}86.05pg/ml$ for saline, B-DC and C-DC treated group, respectively) Natural killer cell activity was also increased in the mice treated with tumor cell lysate pulsed-DC ($8.9{\pm}_{[CGP2]}0.1,\;11.6{\pm}0.8\;and\;12.6{\pm}0.7%$ specific NK activity for saline, B-DC and C-DC treated group, respectively). Conclusion: Conclusively, promising data were obtained that allogeneic-tumor cell lysate can be used as a tumor antigen for DC-based cancer immunotherapy.