Production of Interferon-$\gamma$ from Human Peripheral Blood Lymphocytes

정일엽,염영일,이상기,정태화,한문희,Chung, Il-Yup,Yeom, Young-Il,Rhee, Sang-Ki,Chung, Tai-Wha,Han, Moon-Hi 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3

인체혈액에서 분리한 임파구세포를 PHA-M, Con A, PWM등의 mitogen으로 자극시켜 여러가지 조건하에서 감마-인터페론을 생산하였다. 유도물질로 사용된 T-세포 mitogen인 PHA-M이나 Con A가 B-세포 mitogen 인 PWM보다 감마-인터페론 유도에 훨씬 효율적이었으며 이로부터 임파구세포 중에서 T-세포가 감마-인터페론을 생산하는 주요세포원 임을 확인하였다. 감마-인터페론 생산시 Con A의 적정농도는 $5-10\;{\mu}g/ml$이었다. 유도개시 약 48시간 경과 후 인터페론이 최대로 생산되었고 그 이후에도 인터페론의 활성은 감소되지 않았다. Phorbolester인 TPA를 첨가할 경우 Con A 또는 PHA-M 단독으로 인터페론을 유도시킨 것보다 10배 이상의 감마-인터페론의 생산이 증가되었다. 생산배양액에 첨가되는 혈청의 농도가 인터페론의 생산에 마치는 영향은 매우 극적이어서 태아 송아지 혈청농도 2%에서 최대로 생산되었고 농도가 증가함에 따라 생산역가는 급격히 떨어졌다. 혈청 대신 사용한 알부민 단백질도 비슷한 효과를 나타냈다. 혈액중에 임파구세포에 처리된 Con A가 DNA 합성을 촉진시키는 성질을 이용하여 $^3H$-thymidine 표식으로 cpm을 측정해 본 결과, 이와같은 현상은 10-20%의 높은 혈청농도에서 Con A가 과량의 혈청단백질과 흡착하여 실제적인 Con A 농도를 적정농도 이하로 떨어뜨리기 때문인 것으로 나타났다. Interferon-$\gamma$(IFN-$\gamma$) was produced from human peripheral blood lymphocytes (Hu-PBL) by stimulating them with various kinds of mitogens. T-cell mitogens such as Con A and PHA-M were much more potent than B-cell mitogen such as PWM in producing IFN-$\gamma$ from Hu-PBL. The combined treatment of TP A with these mitogens enhanced IFN-$\gamma$ production ten fold compared with the single treatment of each mitogen. Both DNA synthesis and IFN-$\gamma$ production yielded bell-shaped dose-response curves for a given population of Hu-PBL in response to Con A. The effect of fetal calf serum(FCS) on IFN-$\gamma$ production was found to be critical: titer of IFN-$\gamma$ produced was very low in the absence of FCS and reached peak point at 2% FCS, beginning to decrease precipitously with an increasing FCS concentration up to 20%. Using the relationship between IFN-$\gamma$ production and mitogenic stimulation, it was found that one of factors responsible for the decrease of IFN-$\gamma$ production at the high serum concentration could be the binding of Con A to serum proteins which rendered Con A to be removed from the production system.

Urokinase Immobilized on Cellulose Membrane

김현표,김남득,염영일,변시명,Kim, Hyun-Pyo,Kim, Nam-Deuk,Yeom, Young-Il,Byun, Si-Myung 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.2

인공신장기에 쓰이는 셀룰로오즈 막에 시아노겐 브로마이드방법을 이용한 공유 결합법으로 혈전용해 효소인 유로키나제을 고정화 시켰다. 단순 흡착시킨 유로키나제가 $17-30{\times}10^{-3}$ CTA unit/cm의 활성을 냈으나 계속 사용할 수 없었고 안정성이 전혀 없는 반면, 고정화된 유로키나제는 $19-38{\times}10^{-3}$ CTA unit/cm 의 활성을 보유하였으며, 높은 안정성을 갖고 있는 것으로 판명되어 임상적 사용의 가능성을 높여 주었다. Urokinase was covalently immobilized to a cellulose hollow fiber membrane with the method of CNBr-activation. The resulted immobilized urokinase showed $19-38{\times}10^{-3}$ CTA unit/cm fiber compared to $17-30{\times}10^{-3}$ CTA unit/cm fiber of the simply adsorbed urokinase which showed the least stability during repeated use. The fibrinolytic activity of the immobilized one was very stable and the immobilized fiber promised a potential clinical use.

생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과

정옥(Ok Joung),조영식(Young Sik Cho),조정원(Cheong Weon Cho),이경애(Kyung Ae Lee),심정현(Jung Hyun Shim),조민철(Min Chul Cho),이홍수(Hong Soo Lee),염영일(Young Il Yeom),김상범(Sang Bom Kim),박순희(Sue Nie Park),윤도영(Do Young Yoon) 대한약학회 2000 약학회지 Vol.44 No.4

Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner. In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

Genomics 빅데이터를 이용한 간암 연구

강민호 ( Minho Kang ),염영일 ( Young Il Yeom ) 대한간학회 2016 임상연구방법론워크숍 Vol.2016 No.1

Liver cancer is the fifth most common cancer in men and the seventh in women, and, in developed countries, its incidence is increasing in contrast to most of the other cancer types. The development of hepatocellular carcinoma (HCC), which accounts for 70%~85% of the total liver cancer, is closely associated with multiple risk factors such as chronic infection of hepatitis B (HBV) and C viruses (HCV), alcohol consumption, aflatoxin B1 exposure and obesity. Efforts in large-scale genomic analyses revealed the involvement of diverse oncogenic as well as tumor suppressive genes and pathways in HCC development. However, liver cancer remains highly intractable to anticancer therapies despite the large efforts in clinical trials employing molecularly targeted agents including sorafenib. This is partly due to the tumor heterogeneity that causes the molecular pathogenesis of HCC much complicated. Recent technological advances of high-throughput methods such as the next generation sequencing of cancer genomes and transcriptomes have yielded significant insights into the genetic profiles (somatic mutations, structure variations, HBV integrations, and epigenetic modifications) of HCC. In combination with the high-throughput functional genomics tools such as RNAi and Crispr/Cas9, these genomic informations may make important contributions to identifying biomarkers and therapeutic targets that are critical for the understanding of the pathogenic mechanisms of HCC and the development of individually targeted therapies.

인체 S100A2 단백질에 특이적인 단일클론 항체

김재화,윤선영,김주헌,주종혁,김진숙,이영희,염영일,최용경,최인성,Kim, Jae Wha,Yoon, Sun Young,Kim, Joo Heon,Joo, Jong-Hyuck,Kim, Jin Sook,Lee, Younghee,Yeom, Young Il,Choe, Yong-Kyung,Choe, In Seong 대한면역학회 2003 Immune Network Vol.3 No.1

Background: The S100A2 gene, also known as S100L or CaN19, encodes a protein comprised of 99-amino acids, is a member of the calcium-binding proteins of EF-hand family. According to a recent study, this gene was over-expressed in several early and malignant carcinomas compared to normal tissues. To elucidate the role of S100A2 protein in the process during carcinogenesis, production of monoclonal antibody specific to the protein is essential. Methods: First, cDNA sequence coding for ORF region of human S100A2 gene was amplified and cloned into an expression vector to produce GST fusion protein. Recombinant S100A2 protein and subsequently, monoclonal antibody to the protein were produced. The specificity of anti-S100A2 monoclonal antibody was confirmed by immunoblot analysis of cross reactivity to other recombinant proteins of S100A family (GST-S100A1, GST-S100A4 and GST-S100A6). To confirm the relation of S100A2 to cervical carcinogenesis, S100A2 protein in early cervical carcinoma tissue was immunostained using the monoclonal antibody. Results: GST-S100A2 recombinant protein was purified by affinity chromatography and then fusion protein was cleaved and S100A2 protein was isolated. The monoclonal antibody (KK0723; Korean patent pending #2001-30294) to the protein was produced and the antibody did not react with other members of EF-hand family proteins such as S100A1, S100A4 and S100A6. Conclusion: These data suggest that anti-S100A2 monoclonal antibody produced in this study can be very useful for the early detection of cervical carcinoma and elucidation of mechanism during the early cervical carcinogenesis.

게놈프로젝트란 무엇인가?

염영일 약업신문사 2000 의약정보 Vol.2000 No.8

인체혈 중 임파구로부터 감마 - 인터페론의 생산

이상기,정태화,한문희,염영일,정일엽 생화학분자생물학회 1991 BMB Reports Vol.19 No.3

Interferon-γ (IFN-γ) was produced from human peripheral blood lymphocytes (Hu-PBL) by stimulating them with various kinds of mitogens. T-cell mitogens such as Con A and PHA-M were much more potent than B-cell mitogen such as PWM in producing IFN-γ from Hu-PBL. The combined treatment of TPA with these mitogens enhanced IFN-γ production ten fold compared with the single treatment of each mitogen. Both DNA synthesis and IFN-γ production yielded bell-shaped dose-response curves for a given population of Hu-PBL in response to Con A. The effect of fetal calf serum(FCS) on IFN-γ production was found to be critical: titer of IFN-γ produced was very low in the absence of FCS and reached peak point at 2% FCS, beginning to decrease precipitously with an increasing FCS concentration up to 20%. Using the relationship between IFN-γ production and mitogenic stimulation, it was found that one of factors responsible for the decrease of IFN-γ production at the high serum concentration could be the binding of Con A to serum proteins which rendered Con A to be removed from the production system.

셀룰로오즈 막에 고정화 시킨 유로키나제

김현표,변시명,김남득,염영일 생화학분자생물학회 1991 BMB Reports Vol.19 No.2

Urokinase was covalently immobilized to a cellulose hollow fiber membrane with the method of CNBr-activation. The resulted immobilized urokinase showed 19-38×10^(-3) CTA unit/cm fiber compared to 17-30×10^(-3) CTA unit/㎝ fiber of the simply adsorbed urokinase which showed the least stability during repeated use. The fibrinolytic activity of the immobilized one was very stable and the immobilized fiber promised a potential clinical use.

인체 S100A6 단백질에 특이한 단일클론 항체

김재화,윤선영,김주헌,주종혁,김진숙,이영희,염영일,최용경,최인성 Official Journal of the Korean society for immunol 2003 Immune Network Vol.3 No.1