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종양 미세 환경 내 대식세포에서 혈관 신생 조절 인자로서의 TNF-α에 의한 IFN-γ의 분비 조절
표석능,백소영,곽장동,박대섭,조성준,이현아,Pyo, Suhk-Neung,Baek, Soyoung,Kwak, Jang-Dong,Park, Dae-Sub,Joe, Sung-Jun,Lee, Hyun Ah 대한면역학회 2003 Immune Network Vol.3 No.1
Background: The role of macrophages in tumor angiogenesis is known to be the production of angiogenic cytokines and growth factors including TNF-${\alpha}$. Recently, macrophage also can produce the INF-${\gamma}$ that is being studied to be involved in angiogenic inhibition. Thus, the importance of macrophages in tumor angiogenesis is might being an angiogenic switch. Thus, the hypothesis tested here is that TNF-${\alpha}$ can modulate the INF-${\gamma}$ production in the macrophages from tumor environment as a part of tumor angiogenic switch. Methods: Macrophages in tumor environment were obtained from the peritoneal cavity of C57BL/6 mice injected with B16F10 melanoma cell line for 6 or 11 days. $Mac1^+$-macrophages were purified using magnetic bead ($MACs^{TM}$; Milteny Biotech, Germany) and cultured with various concentrations of TNF-${\alpha}$ for various time points at $37^{\circ}C$. The supernatants were analyzed for IFN-${\gamma}$ or VEGF by ELISA kit (Endogen, Woburn, MA). Results: Residential macrophages from the peritoneal cavity did not respond to LPS or TNF-${\alpha}$ to produce INF-${\gamma}$. However, the cells from tumor environment produced IFN-${\gamma}$ as well as VEGF and upregulated by the addition of LPS or TNF-${\alpha}$. RT-PCR analysis revealed the external TNF-${\alpha}$-induced IFN-${\gamma}$ gene expression in the macrophages from tumor environment. Conclusion: The overall data suggest that the macrophages in tumor environment might have an important role not only in angiogenic signal but also in anti-angiogenic signal by producing related cytokines. And TNF-${\alpha}$ might be a key cytokine in tumor angiogenic switch.
송지영,양현옥,표석능,박신영,김기환,손은화,강남성,윤연숙,Song, Jie-Young,Yang, Hyun-Ok,Pyo, Suhk-Neung,Park, Sin-Young,Kim, Ki-Hwan,Son, Eun-Hwa,Kang, Nam-Sung,Yun, Yeon-Sook 대한약학회 1998 약학회지 Vol.42 No.2
Currently, cancer is the primary cause of death and 50% of cancer patients are incurable by surgery, radiotherapy and chemotherapy. Therefore, immunotherpy is interested as the fourth remedy. Biological response modifier (BRM), such as organometallic compounds, glycoproteins, polysaccharides and other natural products. Is the one which can enhance the immune response against cancer cell. To develop new BRM from natural sources, we investigated 63 species Korean traditional medicines by observing the mitogenic activity to splenocytes, generation of activated killer cells and activation of macrophages. Finally, we selected 9 species including Angelicae gigantis Radix, Mori Cortex Radicis, Arisaematis Tuber, Salviae Radix, Cnidii Rhizoma, Ligusti Fructus, Pasoraliae Semen, Loranthi Ramulus, Ginseng Radix. Bioassay-guided fractionation and purification is undergoing.
민관기(Kwan Kee Min),표석능(Suhk Neung Pyo),이동권(Dong Kwon Rhee) 한국응용약물학회 1999 Biomolecules & Therapeutics(구 응용약물학회지) Vol.7 No.3
Streptococcus pneumoniae (pneumococcus) is the most frequent causative agent of acute bacterial pneumonia. Outstanding characteristic of pneumococcus is an ample polysaccharide capsule that is highly antigenic agent and is the major factor for classification of pneumococcus into more than 94 serotypes. In this study, production of capsular polysaccharide from Streptococcus pneumoniae type 23 was optimized by supplementation of metal ions or by modulation of culture conditions. When brain heart infusion (BHI) broth was supplemented with 1 mM concentration of CaCl₂, CuS0₄ and MgS0₄, growth of pneumococcus as well as polysaccharide yield were stimulated. Also slight aeration gave rise to better polysaccharide yield.
최인화,김수남,김승환,권창희,표석능,이동권,Choi, In hwa,Kim, Su Nam,Kim, Seung Hwan,Kweon, Chang Hee,Pyo, Suhk Neung,Rhee, Dong Kwon 한국미생물학회 1998 미생물학회지 Vol.34 No.1
여러가지 환경 stress에서 유도되는 폐렴구균 열충격단백질 GroEL의 특성에 대하여 검토하였다. 비병원성인 폐렴구균(Streptococcusa pneumoniae)을 사용하여 stress 조건을 설정하고 이 설정된 조건에서 stress에 의해 유도되는 단백질을 $^{35}S$]-methionine으로 표지하여 autoradiography를 실시하였다. 열충격을 가했을 때 유도되는 단백질(65, 73, 84-kDa 등) 중 65 kDa의 단백질(hsp65)을 DEAE-Sepharose ion exchange 및 ATP-agarose affinity chromatography를 이용하여 분리 정제하였으며 hsp65에 대해 생성된 항체를 이용하여 immunoblot을 실시하였을 때 약 60 kDa의 대장균 단백질과 반응하였으며 정제된 폐렴구균 hsp65가 대장균의 anti-GroEL monoclonal 항체와 반응함으로써 폐렴구균 hsp65가 대장균GroEL과 유사한 단백질임을 확인하였다. Induction and purification of the GroEL homolog from Streptococcus pneumolliae were characterized. The stress conditions were determined by temperature, ethanol, NaCI, $H_2O_2$ methyl methanesulfonate, and ethyl methanesulfonate. And stress induced proteins were analyzed using [$^{35}S$]-methionine labeling method. Heat shock induced the synthesis of a set of about 3 heat shock proteins (hsps) (65, 73, and 84-kDa). Of those 3 hsps, a 65 kDa protein, hsp65, was purified by DEAE-Sephacel and ATP-agarose column chromatography, and used for antibody preparation. Immunoblot analysis employing antisera raised against pneumococcus hsp65 demonstrated cross-reactivity with a 60 kDa protein in Eschericha coli. Also cross reaction of the purified p65 with anti-Escherichia coli GroEL monoclonal antibody demonstrated that pneumococcal hsp65 is the GroEL homolog.
손은화,박재현,김경란,김병오,이동권,표석능,Son, Eun-Wha,Park, Jae-Hyun,Kim, Kyung-Ran,Kim, Byung-O,Rhee, Dong-Kwon,Pyo, Suhk-Neung 한국생약학회 1998 생약학회지 Vol.29 No.1
The effects of Bezoar Bovis on immune responses of ICR Mice were studied. In the present study, the Jerne hemolytic plaque assay (PFC) was used to evaluate the humoral immune response to sheep red blood cells, and macrophage regulatory function was measured by quantitating the production of molecules secreted by macrophages. Bezoar Bovis was given in single daily p.o doses for short term (1, 2 days) or long term (7, 14 days). PFC response for taken 3 days after short term exposure to Bezoar Bovis was increased. The production of nitirc oxide in macrophages was also stimulated. In contrast, long term exposure to Bezoar Bovis resulted in inhibitory effect of Bezoar Bovis on nitirc oxide, $TNF-{\alpha}$and IL-6 production in macrophages. These findings suggest that treatment with Bezoar Bovis may result in differential immunological effect depending on treatment schedule.
김경란(Kyung Ran Kim),조대현(Dae Hyun Cho),표석능(Suhk Neung Pyo) 한국응용약물학회 1999 Biomolecules & Therapeutics(구 응용약물학회지) Vol.7 No.2
Triazine herbicide has been reported to directly suppress the immune response. In the present study, the effect of simazine on the immune response was investigated. Splenic lymphocytes were treated withmitogen (lipopolysaccaride, concanavalin A) in the presence of simazine. When simazine(300 mg/kg, 600 mg/kg) was administrated every day for 3 weeks or 4 weeks, respectively, the number of plaque forming cells (PFC) was decreased. Antibody production of IgM and IgG class was significantly decreased in splenic cells from simazine-treated animals. In addition, when animals were exposed to simazine, the susceptibility of virus infection as well as the growth of tumor cells was increased. These data suggest that simazine affected the immune function and humoral immunity impaired by simazine treatment contributed to pathological process.
이동권(Dong Kwon Rhee),문은이(Eun Yi Moon),표석능(Suhk Neung Pyo) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.2
Aflatoxin B1 (AFB1) has been reported to directly suppress the immune responses. In the present study, the effect of AFB1 on immune functions was investigated. Splenic lymphocytes were treated with various doses of the mitogens (lipopolysaccharide, concanavalin A) in the presence of AFB1. AFB1 pretretment decreased the number of plague forming cells (PFC) in a dose-dependent manner. Antibody production of IgM and IgG class was significantly decreased in AFB1-treated splenic cells. In addition, when animals were exposed to AFB1, the susceptibility of bacterial infection as well as the growth of tumor cells was increased. These data suggest that AFB1 affected the immune function and humoral immunity impaired by AFB1 treatment contributed to pathological process.
84 - kDa 의 폐렴구균 열충격단백질 CipL 의 Cloning 및 면역특성에 관한 연구
권혁영(Hyog Young Kwon),김용환(Yong Hwan Kim),최혜진(Hye Jin Choi),박연진(Youn Jin Park),표석능(Suhk Neung Pyo),이동권(Dong Kwon Rhee) 한국응용약물학회 2001 Biomolecules & Therapeutics(구 응용약물학회지) Vol.9 No.2
Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal C1pL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clp member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneumococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.