• 식물조직배양학회지
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• 제21권6호
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• pp.353-356
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• 1994

사람 B형 간염 바이러스의 pre-S2 표면항원에 결합하는 키메라 항체 유전자(카파 및 감마사슬의 cDNA클론)를 식물체에서 발현시키기 위해 식물체 발현벡터인 pBKS-1에 XbaI 자리를 이용하여 클로닝하였다. 이들 유전자를 포함하는 대장균의 플라스미드 핵산을 추출하여 아그로박테리움에 형질전환 시켰다. 다음 담배의 조직절편과 아그로박테리움을 공동배양함으로써 담배의 형질 전환을 시도하였다. 카나마이신이 포함된 신초유기배지에서 나온 신초를 시료로 하여 Western blot을 실시함으로써 이들 유전자가 형질전환 담배에서 안정하게 발현됨을 확인하였다.

• Childhood Kidney Diseases
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• 제24권1호
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• pp.1-13
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• 2020

Immunoglobulin (Ig)A vasculitis nephritis (IgAVN), also referred to as Henoch-Schönlein purpura nephritis, is a relatively benign disease in children. However, two 24-year European cohort studies have reported high sustained rates of hypertension, severe proteinuria, and renal dysfunction in patients with IgAVN. Notably, the incidence and exacerbation rates of proteinuria, hypertension, and renal dysfunction during pregnancy were high even in women who recovered from IgAVN before pregnancy. Patients with IgAVN need lifelong care. Trials have been performed to investigate early biomarkers and genes associated with poor prognosis to identify high-risk patients in whom IgAVN may progress to severe renal disease. Urinary IgA/cr, IgM/cr levels, and HLAB35 and angiotensinogen gene expression were shown to be predictors of progression of IgAVN to severe renal dysfunction. The 2019 Single Hub and Access point for paediatric Rheumatology in Europe (SHARE) initiative group published guidelines for pediatric IgAVN, following the Kidney Disease: Improving Global Outcomes (KDIGO) guidelines established in 2012. Compared with the KDIGO guidelines, the SHARE guidelines recommend earlier corticosteroid administration in cases of mild proteinuria (>0.5 g/d). Clinical trials of targeted budesonide delivery to the distal ileum, monoclonal antibody targeting C5, eculizumab and anti-CD20 monoclonal antibody administration, among others are currently underway in patients with IgA nephropathy. It is expected that newer therapeutic agents would become available for IgAVN in the near future. This review summarizes IgAVN with emphasis on recently published literature, including possible preventive strategies, predictive biomarkers for progression of IgAVN, and various treatments.

• 대한본초학회지
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• 제30권1호
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• pp.25-32
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• 2015

Objectives : In this study, we investigated the effect of Cassia obtusifolia Linne (Cassiae Semen; CS) extract on ovalbumin (OVA)-induced allergic asthma in mice. Methods : CR was extracted with 70% ethanol. For in vitro study, HMC-1, human mast cells were treated with CS extract at 0.2 and $0.5mg/m{\ell}$ for 1 h, and then stimulated with compound (C) 48/80 for 30 min. Primary spleenocytes were isolated from the spleen of mice, treated with CS extract for 1 h, and then stimulated with ConA for 24 h. For in vivo study, mice were sensitized at day 0, 7 and 14 with 0.2% OVA and then airway challenged using neublizer at day 21, 23, 25, and 27 to induced allergic asthma. CS extract at doses of 100 and 300 mg/kg body weight was orally administered during OVA challenge once per a day. The levels of allergic mediators such as histamine, OVA-specific IgE, IL-4, and $IFN-{\gamma}$ were measured in the sera of mice or culture supernatants by EIA and ELISA, respectively. The expression of IL-4 and $IFN-{\gamma}$ gene was determined by RT-PCR. The histopathological change of lung tissues was observed with hematoxylin and eosin (H&E) and Periodic acid Schiff (PAS) staining. Results : The treatment of CS extract in HMC-1 cells significantly inhibited C48/80-induced degranulation, and histamine release. The treatment of CS extract in spleenocytes suppressed the expression of IL-4 and $IFN-{\gamma}$ mRNA. The administration of CS extract in OVA-induced asthmatic mice significantly decreased the levels of OVA-specific IgE, and IL-4 in a dose-dependent manner with OVA-control group. In addition, CS extract inhibited the infiltration of inflammatory cells and bronchiolar damage with epithelial thickening in lung tissues of OVA-induced asthma mice, and also mucin accumulation. Conclusions : These results indicate that CS extract prevents asthmatic damage through regulating the allergic immune response.

• 생약학회지
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• 제51권4호
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• pp.340-348
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• 2020

This study was conducted to evaluate the immunomodulatory effects of red doraji (Platycodon grandiflorum, RD) prepared by repeated steaming and drying process in the immune-suppressed mice induced by pre (RD-A) or post-treatment (RD-B) with cyclophosphamide. The immune-stimulating effects of ethanol RD extract in in vivo at 150 (RD-1) and 300 mg/kg body weight (RD-2) for RD-A and RD-B groups were measured and compared to the NC group supplied with distilled water only or positive control group. After 14 days of oral supplement, serum IgA, IgG, and cytokine levels, splenocytes proliferation rate, NK cell activity, and gene expression of cytokines were measured as immune related biomarkers. Serum IgA, IgG, IL-1β, and IL-12 levels increased in both RD-A and RD-B groups while serum TNF-α level decreased in RD-A group compared to the NC group. Splenocytes proliferation rate, NK cell activity, and cytokine (IL-1β, IL-6, IFN-γ) expression levels were also improved by RD supplement in the both groups. The RD showed more significant immunomodulatory effects at higher dose (RD-2) rather than the lower dose (RD-1). Thus, RD has an immune efficacy in a dose dependent manner and can be used as an immune stimulating source to improve immunity.

• Journal of Ginseng Research
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• 제46권2호
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• pp.304-314
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• 2022

Background: Ginsenosides are biologically active components of ginseng and have various functions. In this study, we investigated the immunomodulatory activity of a ginseng product generated from ginseng powder (GP) via enzymatic bioconversion. This product, General Bio compound K-10 mg solution (GBCK10S), exhibited increased levels of minor ginsenosides, including ginsenoside-F1, compound K, and compound Y. Methods: The immunomodulatory properties of GBCK10S were confirmed using mice and a human natural killer (NK) cell line. We monitored the expression of molecules involved in immune responses via enzyme-linked immunosorbent assay, flow cytometry, NK cell-targeted cell destruction, quantitative reverse-transcription real-time polymerase chain reaction, and Western blot analyses. Results: Oral administration of GBCK10S significantly increased serum immunoglobulin M levels and primed splenocytes to express pro-inflammatory cytokines such as interleukin-6, tumor necrosis factor-α, and interferon-γ. Oral administration of GBCK10S also activated NK cells in mice. Furthermore, GBCK10S treatment stimulated a human NK cell line in vitro, thereby increasing granzyme B gene expression and activating STAT5. Conclusion: GBCK10S may have potent immunostimulatory properties and can activate immune responses mediated by B cells, Th1-type T cells, and NK cells.

• Pediatric Infection and Vaccine
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• 제29권3호
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• pp.166-172
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• 2022

블라우증후군(Blau syndrome)은 nucleotide-binding oligomerization domain protein 2 (NOD2) 유전자의변이에 의해 발생하는 질환으로 육아종성 피부염 및 만성 포도막염, 관절염을 특징으로 한다. 증상이 비특이적이고 동시다발적으로 발생하지 않아 진단이 어려운 경우가 많다. 반복되는 피부 발진 및 발열에 대해 두 차례 가와사키병으로 오진되어 면역글로불린과 전신 스테로이드로 치료받은 바 있는 13개월 남자 환자에서 블라우 증후군을 진단한 증례를 보고하고자 한다.

• IMMUNE NETWORK
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• 제20권5호
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• pp.42.1-42.10
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• 2020

Long-lasting post-switched plasma cells (PCs) arise mainly from germinal center (GC) reactions, but little is known about the mechanism by which GC B cells differentiate into PCs. Based on our observation that the expression of the transcription factor CCAAT/enhancer binding protein β (C/EPBβ) is associated with the emergence of post-switched PCs, we enquired whether a cell-autonomous function of C/EPBβ is involved in the program for PC development. To address this, we generated C/EPBβ-deficient mice in which the Cebpb locus was specifically deleted in B cells after transcription of the Ig γ1 constant gene segment (Cγ1). In response to in vitro stimulation, B cells from these Cebpb^fl/flCγ1^Cre/+ mice had defects in the induction of B lymphocyte-induced maturation protein 1 (Blimp1) and the formation of IgG1⁺ PCs, but not in proliferation and survival. At steady state, the Cebpb^fl/flCγ1^Cre/+ mice had reduced serum IgG1 titers but normal IgG2c and IgM titers. Moreover, upon immunization with T-dependent Ag, the mice produced reduced levels of Ag-specific IgG1 Ab, and were defective in the production of Ag-specific IgG1 Ab-secreting cells. These results suggest that a cell-autonomous function of C/EPBβ is crucial for differentiation of post-switched GC B cells into PCs through a Blimp1-dependent pathway.

• 대한임상검사과학회지
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• 제48권4호
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• pp.371-377
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• 2016

탈회방법은 골수조직의 병리학적 진단을 위해서 항상 시행되는 과정이다. HCl 탈회용액과 같이 주로 사용하고 있는 산성용액은 탈회과정 동안에 조직내의 항원성에 손상을 입힌다. 특히, 골수조직 내의 RNA나 DNA에 심하게 손상을 준다. 따라서 조직의 항원성을 보존하기 위한 표준화된 탈회방법이 필요하다. 본 연구는 일반적으로 가장 많이 사용되는 HCl 기반의 상품화된 탈회용액과 직접 제조한 EDTA 탈회용액이 골수조직의 탈회과정에 어떤 영향을 미치는지 분석하였다. 환자로부터 채취된 73예의 골수생검조직을 HCl 탈회와 EDTA 탈회의 두 그룹으로 나누어 탈회과정을 진행하였다. 골수생검조직의 탈회과정 후 결과의 차이는 hematoxylin & eosin 염색과 reticulum 염색, Ki-67, CD20, CD138의 항체를 이용한 면역조직화학염색, DNA 추출 및 분석, in situ hybridization, IGH gene rearrangement 와 같은 분자병리검사를 시행하여 분석하였다. 일반적인 염색과 특수염색에서는 두 탈회용액간의 차이는 없었다. 또한 세포증식 표지자와 같은 세포막 혹은 세포질에서 발현되는 항체는 탈회용액간의 차이 없이 잘 염색되었다. 반면 HCl 탈회 용액에 처리한 후 핵 내 단백질인 Ki-67의 염색상은 현저히 불량한 것으로 관찰되었다. HCl 탈회용액과 비교하여 EDTA 탈회용액에서의 골수생검조직 내의 DNA와 RNA가 잘 보존되었음을 다양한 분자병리검사를 통해 확인할 수 있었다. 특히 HCl 탈회용액에 처리한 28예와 EDTA 탈회용액에 처리한 12예의 DNA의 순도와 농도을 비교한 결과 통계학적으로 유의한 수준으로 차이가 있음을 확인하였다. 이로써 EDTA 탈회용액이 조직 내의 항원성을 잘 유지시키며, 면역조직화학염색과 분자병리검사에 적합한 방법임을 확인 할 수 있었다.

• BMB Reports
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• 제39권6호
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• pp.696-702
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• 2006

Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/macrophage colony-stimulating factor and FMS-like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno-fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters.

• 대한핵의학회지
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• 제35권5호
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• pp.313-323
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• 2001

목적: 저선량 방사선에 의해 그 이후의 고선량 방사선에 저항이 생기는 유익한 반응을 보인다는 방사선적응반응이라는 현상이 알려져 있지만, 저선량 방사선이 어떤 기작에 의해 이런 반응을 일으키는지에 대해서는 아직 알려지지 않고 있다. 본 연구에서는 정상 인체세포주에서 저선량 $^{99m}Tc$에 의해 방사선적응반응이 유도되는지를 확인하고, 이 때 활성화되는 유전자를 찾아보고자 하였다. 대상 및 방법: 인체 정상 림프구 세포주인 NC-37 세포주 $2{\times}10^6mL$개의 세포에 $^{99m}Tc$을 148 MBq/mL로부터 148 Bq/mL의 농도가 되도록 10배씩 희석하여 첨가하고 44시간동안 배양하였다. 결과: 각각의 군에 대해 이상 염색체를 계수하여 148 KBq/mL의 $^{99m}Tc$을 첨가한 군에서 방사선적응반응이 가장 현저하게 유도되었음을 확인하였다. 이 세포군에서 mRNA를 추출하고 여기에서 cDNA를 만든 후 gene discovery array (GDA) 여과기를 이용하여 대조군에 비해 발현이 증가된 casein kinase II beta chain, immunoglobulin, HLA-B 그리고 아직 알려지지 않은 2개의 유전자 등 6개의 유전자를 찾아내었다. Representational difference analysis (RDA)법을 통해서는 대조군에 비해 발현이 증가된 유전자 클론을 20개 찾아내었다. 결론: 인체세포주 NC-37에서 저선량의 $^{99m}Tc$에 의해 방사선적응반응이 유도된다는 사실을 밝혔으며, 이때 다수의 유전자가 발현된다는 사실을 알 수 있었다.