• Journal of Nutrition and Health
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• v.43 no.2
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• pp.152-160
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• 2010

The purpose of this study was to investigate the effects of aging process on the immunity in human subjects. In this investigation, nineteen families of three generations (daughters on college age, their mothers, and grandmothers) participated to avoid genetic variation among individuals. Dietary food records, anthropometric measurements and biochemical assessments of serum nutrients were used to evaluate the nutritional status of subjects. The immune parameters of subjects were assessed by the total and differential WBC count. Total B and T lymphocytes, and T cell subsets were quantified by flowcytometer. Serum immunoglobulin G, A, M concentrations were also measured as an index of humoral immunity. The result of this study can be summarized as follows: 1. Along with the aging process, body fat was found to be increased whereas lean body mass and total body water were diminished. Since there were no significant difference in serum vitamin E levels in all age groups, serum retinal concentrations tended to decrease as one gets old. 2. Although total number of T lymphocytes seemed to be unchanged, B lymphocytes and NK cell numbers were increased by aging. The Percentage of CD8 + lymphocytes was lower in the elderly subjects compared with the younger, resulting in higher ratio of CD4 +/CD8 + lymphocytes in the elderly. Serum Ig G and Ig A levels remained unchanged, but IgM levels were significantly decreased as the age processes continue. Taking all together, it could be suggested that the alteration of immune cell population by aging is selective and possibly nonage factors such as nutrition may be attributable to the change of immunity in the elderly. The nutritional status and aging process may selectively affect both the cell-mediated (CD8 +, CD4 + CD8 + ratio, NK cell) and humoral (B lymphocyte, Immunoglobulin M, G) immune parameters in human subjects.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.651-657
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• 2019

Although smallpox was eradicated in 1980, it is still considered a potential agent of biowarfare and bioterrorism. Smallpox has the potential for high mortality rates along with a major public health impact, eventually causing public panic and social disruption. Passive administration of neutralizing monoclonal antibodies (mAbs) is an effective intervention for various adverse reactions caused by vaccination and the unpredictable nature of emerging and bioterrorist-related infections. Currently, vaccinia immune globulin (VIG) is manufactured from vaccinia vaccine-boosted plasma; however, this production method is not ideal because of its limited availability, low specific activity, and risk of contamination with blood-borne infectious agents. To overcome the limitations of VIG production from human plasma, we isolated two human single-chain variable fragments (scFvs), (SC34 and SC212), bound to vaccinia virus (VACV), from a scFv phage library constructed from the B cells of VACV vaccine-boosted volunteers. The scFvs were converted to human IgG1 (VC34 and VC212). These two anti-VACV mAbs were produced in Chinese Hamster Ovary (CHO) DG44 cells. The binding affinities of VC34 and VC212 were estimated by competition ELISA to $IC_{50}$ values of $2{\mu}g/ml$ (13.33 nM) and $22{\mu}g/ml$ (146.67 nM), respectively. Only the VC212 mAb was proven to neutralize the VACV, as evidenced by the plaque reduction neutralization test (PRNT) result with a $PRNT_{50}$ of ~0.16 mg/ml (${\sim}1.07{\mu}M$). This VC212 could serve as a valuable starting material for further development of VACV-neutralizing human immunoglobulin for a prophylactic measure against post-vaccination complications and for post-exposure treatment against smallpox.

• Clinical and Experimental Vaccine Research
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• v.12 no.4
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• pp.328-336
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• 2023

Purpose: Human peripheral blood mononuclear cell (PBMC)-based in vitro systems can be of great value in the development and assessment of vaccines but require the right medium for optimal performance of the different cell types present. Here, we compare three commonly used media for their capacity to support innate and adaptive immune responses evoked in PBMCs by Toll-like receptor (TLR) ligands and whole inactivated virus (WIV) influenza vaccine. Materials and Methods: Human PBMCs were cultured for different periods of time in Roswell Park Memorial Institute (RPMI), Dulbecco's minimal essential medium (DMEM), or Iscove's modified DMEM (IMDM) supplemented with 10% fetal calf serum. The viability of the cells was monitored and their responses to TLR ligands and WIV were assessed. Results: With increasing days of incubation, the viability of PBMCs cultured in RPMI or IMDM was slightly higher than that of cells cultured in DMEM. Upon exposure of the PBMCs to TLR ligands and WIV, RPMI was superior to the other two media in terms of supporting the expression of genes related to innate immunity, such as the TLR adaptor protein gene MyD88 (myeloid differentiation factor 88), the interferon (IFN)-stimulated genes MxA (myxovirus resistance protein 1) and ISG56 (interferon-stimulated gene 56), and the leukocyte recruitment chemokine gene MCP1 (monocyte chemoattractant protein-1). RPMI also performed best with regard to the activation of antigen-presenting cells. As for adaptive immunity, when stimulated with WIV, PBMCs cultured in RPMI or IMDM contained higher numbers of IFNγ-producing T cells and secreted more immunoglobulin G than PBMCs cultured in DMEM. Conclusion: Taken together, among the different media assessed, RPMI was identified as the optimal medium for a human PBMC-based in vitro vaccine evaluation system.

• Neonatal Medicine
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• v.25 no.2
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• pp.58-65
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• 2018

Purpose: The purpose of this study is to describe the rate of cytomegalovirus (CMV) virolactia, and the prevalence of breast milk (BM)-transmitted postnatal CMV infection among premature infants after freeze-thawing (FT) and Holder pasteurization (HP) of breast milk. Methods: This is a single-center, retrospective study of 312 infants born at less than 32 weeks of gestation, or with a birth weight less than 1,500 g from January 2013 to June 2017. All infants were screened for CMV-specific immunoglobulin (Ig) G and IgM at birth. Initial CMV specific polymerase chain reaction (PCR) and CMV culture were performed on mothers' BM and babies' urine within the first 21 days of life. FT and HP of BM was used to prevent the transmission of CMV. For the surveillance of postnatal CMV infection, CMV culture and CMV specific PCR of urine from babies were repeated one to two months after the initial screening. Screening for viremia and viruria was performed if postnatal CMV infection was suspected. Results: Among 178 BM samples obtained from mothers of CMV-IgG-seropositive infants, 80 (44.9%) were CMV PCR positive. CMV deoxyribonucleic acid (DNA) was detected in five of the 22 BM samples (22.7%) obtained from the mothers of CMV-IgG seronegative infants. When CMV DNA load in BM was measured before and after HP, various results were shown. Sixty-three infants out of 232 (27.2%) were evaluated for postnatal CMV infection and four infants out of 63 (6.3%) were infected. Conclusion: Interventions to prevent BM-transmitted CMV infection can reduce the chance of postnatal CMV infection, but not completely eliminate it.

• Reproductive and Developmental Biology
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• v.36 no.2
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• pp.133-139
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• 2012

This study aims to examine the effect of Genetically Modified ${\beta}$-Carotene Biofortified Rice rice developed by simultaneous expression technology in NAAS on biological immunity. Accordingly, this study added Genetically Modified ${\beta}$-Carotene Biofortified Rice 25, 50% and general rice 50% as control group into diet and provided rats with the prescribed feeds and then measured the contents of immunoglobulin and cytokine in blood. As a result, male and female IgM, IgE, male IgG1, female IgG2a and TNF-a, IL5 and IL12 showed no significant difference; male IgG2a tended to decrease dependently on the combined concentration of Genetically Modified ${\beta}$-Carotene Biofortified Rice; female IgG1 showed significance with control group, but its association with diet was not found. The higher the dietary mixing ratio, the more the male and female IFN-a and female IL-4 contents, regardless of rice variety, and it was found that female IL6 content decreased significantly, but its association with diet was not found. The risk of beta carotene-enriched rice into environment and human body has not been reported yet. The digestion of Genetically Modified ${\beta}$-Carotene Biofortified Rice can be seen as "safe" as this test result showed no big difference between general rice and Genetically Modified ${\beta}$-Carotene Biofortified Rice, and its usability is full of suggestions.

• Preventive Nutrition and Food Science
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• v.22 no.1
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• pp.9-15
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• 2017

Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor ($Fc{\varepsilon}RI$) ${\alpha}$ chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be $170{\pm}1.9mg$ gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of $Fc{\varepsilon}RI$ was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the $Fc{\varepsilon}RI$ ${\alpha}$ chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of $Fc{\varepsilon}RI$ and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([$Ca^{2+}$]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of $Fc{\varepsilon}RI$ expression through inhibition of ERK 1/2 activation.

• Journal of Pharmacopuncture
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• v.7 no.3
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• pp.5-25
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• 2004

Objectives : To observe changes in the serum proteins before and after intravenous injection of wild ginseng herbal acupuncture. Methods : Blood was collected before and after the administration of wild ginseng herbal acupuncture and only the serum was centrifuged. Then differences in the spots on the scanned image after running 2-Dimensionl electrophoresis were located and conducted mass analysis and protein identification. Results : Following results were obtained from the comparative analysis of serum proteins before and after the administration of wild ginseng herbal acupuncture. 1. 28 spots were identified before and after the administration. 2. In confirming manifestation degree, spots with more than two-times increase were 204, 803, 1505, 2205, 3105, 7104, 9001 spots, with more than one-time increase were 1101, 1302, 2013, 3009, 3010, 4002, 4009, 6706, 7103, 8006, 8101, and spots with decrease were 205, 801, 3205, 5202, 6105. 3. After conducting protein identification, proteins 205, 804, 1302, 4009, 6105, 6106 are unidentified yet, and 1101 is unnamed protein. Protein 204 is identified as complement receptor CR2-C3d, 801 as YAP1 protein, 803 as antitrypsin polymer, 1505 as PRO0684, 2013 and 3010 as proapolipoprotein, 2205 as USP48, 2403 as vitamin D binding protein, 3009 as complement component 4A preprotein, 3105 as immunoglobulin lambda chain, 3205 as transthyretin, 4002 as Ras-related protein Ral-A, 4204 as beta actin, 5202 and 7104 as apolipoprotein L1, 6704 as alpha 2 macroglobulin precursor, 7103 as complement component 3 precursor, 8006 as testis-specific protein Y, 8101 as Transferrin, 9001 as(Alpha-Oxy, Beta-(C112g)deoxy) T-State Human Hemoglobin, and 9003 as human hemoglobin. 4. Immune protein CR2-C3d, which acts against microbes and pathogenic organisms, and Antitrypsin(803), which is secreted with inflammatory response in the lungs, were increased by more than 200% after the administration of herbal acupuncture. 5. Immunoglobulin lambda chain(3105), Alpha-Oxy, Beta-(C112g)deoxy T-State Human Hemoglobin(9001), and human hemoglobin(9003) were increased by more than two-times after the administration of herbal acupuncture. 6. Proapolipoprotein(2013, 3010) and apolipoprotein(7104), key components of the HDL-cholesterol which plays an important role in preventing arteriosclerosis, were increased after the administration of herbal acupuncture. 7. Vitamin D binding protein(DBP, 2403), protecting the lung at the time of inflammatory response, was increased after the administration of herbal acupuncture. 8. Transthyretin(TTR, 3205), which is the main protein causing familial aimyloid polyneuropathy(FAP), was decreased after the administration of herbal acupuncture. 9. Ras-related protein Ral-A(4002) that controls phospholipid metabolism, cytoskeletal formation, and membrane traffic, was increased after the administration of herbal acupuncture. 10. Testis-specific protein Y(8006), which takes part in determination of the gender, was increased by more than two-times after the administration of herbal acupuncture. 11. Transferrin(8101), T-State Human Hemoblobin(9001), and Human Hemoblobin(9003) which balances the iron level in the body, were increased after the administration of herbal acupuncture. Conousion : Above results support the notion that intravenous injection of cultivated wild ginseng herbal acupuncture induce changes in serum proteins and this research can be a pioneer work in finding biomarkers.

• Tuberculosis and Respiratory Diseases
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• v.42 no.4
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• pp.455-464
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• 1995

Background: Considering that both humoral and cell mediated immunities play an important role for human tuberculosis infection, enzyme-linked immunosorbent assay(ELISA) measurement of immunoglobulin G (IgG) antibody to mycobacterial antigens can be used for the serologic diagnosis of tuberculous pleural effusion. Method: We measured absorbance values of IgG antibodies to purified-protein-derivative (PPD) and lipoarabinomannan-B (LAM-B) in the pleural fluid (PF) and the serum in 40 tuberculous (TPE) and 19 nontuberculous pleural effusions (NTPE). Results: 1) The IgG antibodies to PPD and LAM-B were significantly (P<0.0005) higher in the PF and the serum of TPE compared to NTPE. 2) The IgG antibodies to PPD and LAM-B in the serum were higher than that in PF. 3) Significant correlations were found between pleural and serum IgG antibodies to PPD and LAM-B. 4) With a cutoff value for IgG antibody to PPD in the PF of 0.091, sensitivity was 55.0% and specificity 94.7% in the diagnosis of TPE. 5) With a cutoff value for IgG antibody to LAM-B in the PF of 0.337, sensitivity was 50.0% and specificity 94.7% in the diagnosis of TPE. 6) The seropositive rates in TPE were not related to PPD skin test status, the amount of PF and coexisting active pulmonary tuberculosis. Conclusion: The assay of IgG antibodies to PPD and LAM-B might be useful for the diagnosis of TPE. Our study suggests the mechanism of passive transfer of IgG antibodies to PPD and LAM-B from the serum to the PF through pleural tissue.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.81-89
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• 2004

Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.511-519
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• 2009

A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.