Vinblastine Determination Measured by a Sensitive ELISA Inhibition Assay

Jae-Wha Kim(김재화),Mi-Young Han(한미영),Hee-Gu Lee(이희구),Eun-Young Song(송은영),Tai-Wha Chung(정태화),Kyung-Soo Nam(남경수),In-Seong Choe(최인성),최용경 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

Vinblastine을 포함하는 bis-indole alkaloids에 대한 단일클론 항체를 생산하여 Vinca alkaloids의 양을 측정할 수 있는 간편한 immunoassay 체계를 확립하였다. Vinca alkaloids는 periwinkle식물체의 배양된 세포로부터 추출하여 BSA와 접합한 후 Balb/c생쥐에 면역시켜 얻은 비장세포와 골수종양세포의 융합을 유도하여 VBL-BSA에 반응하는 클론을 ELISA 방법으로 분석하였으며 이들 클론 중 bis-indole alkaloids와 특이적으로 반응하는 항체는 inhibition assay를 통하여 분리할 수 있었고 그 결과 두 개의 단일클론 항체를 형성하는 세포주(KN-1과 KN-2)를 확립하였다. KN-1의 경우 dimeric bis-indole alkaloids와는 상당한 교차반응을 나타낸 반면 monomeric bis-indole alkaloids와는 교차반응을 나타내지 않았으며 이 클론의 항체를 이용하여 배양된 세포 추출물에 포함된 Vinca alkaloids의 양을 측정한 결과 0.05 nM정도의 dimeric Vinca alkoloids까지도 측정할 수 있었다. Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay). These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

미립자 응집반응을 이용한 C-reactive Protein의 면역측정법에 관한 연구

김재화(Jae-Wha Kim),송은영(Eun-Young Song),이희구(Hee-Gu Lee),최용경(Yong-Kyung Choe),최명자(Myung-Ja Choi),김용호(Yong Ho Kim),최인성(In-Seong Choe),정태화(Tai-Wha Chung) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

환자의 복수와 늑막액으로부터 p-diazonium phenylphosphorylcholine(DPPC) coupled Separose-4B affinity chromatography와 hydroxylapatite chromatography를 실시하여 C-reactive protein(CRP)를 분리, 제정하였다. 정제된 CRP를 토끼에게 면역화하여 항혈청을 얻고 affinity chromatography를 하여 면역항체(IgG)를 분리하였다. 분리된 면역항체를 미립자에 감작시킨 후 미립자 응집반응에 의하여 3분내에 CRP를 측정할 수 있는 간이 면역측정법을 개발하였다. 본 연구에서 개발된 CRP측정법의 검출범위는 0.5~20㎎/dl이며, 임상 시험 결과 0.7~2.9㎎/dl에서는 강한 응집반응을, 5.0~13.2㎎/dl에서는 약한 응집반응을 보였고 28㎎/dl이상에서는 항원 과잉으로 인한(zone of Ag excess phenomenon) 위음성을 나타냈다. 74명의 환자 혈청을 대상으로 CRP의 농도를 조사한 결과 평균치는 3.8㎎/dl이었으며 대부분의 환자에서는 10㎎/dl 이하의 농도로 존재하였다. 그러므로 1차판정시 음성을 나타낸 시료라도 혈청을 5~10배정도 희석하여 재분석한다면 오차없이 CRP를 검출할 수 있었다. 환자 혈청을 검체로 하여 본 연구에서 개발한 면역측정법과 현재 수입 시판중인 프랑스의 B사 제품과 일본의 I사 제품을 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가분석을 통하여 볼 때 본 연구에서 개발한 간이 면역측정법은 사용이 비교적 간편하며 신빙성이 있어 CRP를 스크리닝 하는데 효과적임을 알 수 있었다. The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5㎎/dl and 20㎎/dl in serum, showing strong response in the range of 0.7~2.9 ㎎/dl, weak response in 5.0~13.2 ㎎/dl and zone phenomenon over 28㎎/dl. The average value of CRP in 74 samples was 3.8㎎/dl and most of the values were lower than 10㎎/dl. The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed mi-croparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

조각난 경관에서 멸종위기종 붉은점모시나비의 서식지 패치 네트워크 분석과 보전

김도성,박성준,조영호,김기동,도재화,서형수,신영규,서민환,오길종,Kim, Do-Sung,Park, Seong-Joon,Cho, Young-Ho,Kim, Ki-Dong,Tho, Jae-Wha,Seo, Hyung-Soo,Shin, Young-Kyu,Suh, Min-Hwan,Oh, Gil-Jong 한국응용곤충학회 2012 한국응용곤충학회지 Vol.51 No.1

종에 대한 생태적 특성과 서식지에 대한 이해는 종의 보전에 매우 중요하다. 본 연구는 멸종위기에 처해있는 붉은점모시나비의 생태적 특성을 바탕으로 서식지 패치네트워크를 분석하였다. 그 결과 포획 개체수는 188개체, 재포획은 220회 되었다. 그리고 암수의 비율은 42:146개체로 암컷보다는 수컷이 약 4배 많은 것으로 나타났다. 또한 개체의 평균생존일수는 $3.93{\pm}3.93$일(수컷: $4.0{\pm}3.9$, 암컷: $2.5{\pm}1.0$), 암컷과 수컷의 최대 생존일수는 각각 13, 14일 나타났고, 수컷이 암컷에 비하여 오래 생존하는 개체가 많은 것으로 나타났다. 종의 평균이동거리는 377 m을 보였으며 최대 1550 m까지 이동하는 것으로 나타났다. 패치연결성과 개체생존이주율의 추정에서 패치간의 거리가 약 300 m 이내가 종의 이주에 적합하며 600 m 이상 떨어질 경우 개체생존이주율이 급격하게 감소하는 것으로 나타났다. 또한 종의 이주 빈도는 근접한 거리에서 다수의 패치가 있는 곳에서 활발하게 일어나고 있어 종의 보전을 위해서는 근접한 거리에 다수의 패치가 필요함을 알 수 있었다. 이번 연구 결과는 붉은점모시나비의 서식지 특성이 분석되어 종 보전을 위한 서식지 디자인 및 설계에 유용하게 사용될 수 있을 것으로 본다. Understanding the ecological complexity and habitat of a species are crucially important to conserve an endangered species. This study evaluated the patch network ecology of the endangered species $Parnassius$ $bremeri$. The results indicated that 188 individuals were captured and 220 were recaptured, respectively. The sex ratio of female: male was 42:146; males were four times more abundant than females. The average longevity of an adult was $3.93{\pm}3.93$ days (male, $4.0{\pm}3.9$; female, $2.5{\pm}1.0$ days); the maximum longevity was 14 days for males and 13 days for females, respectively. Therefore, the expected longevity of males was longer than that of females. The average emigration distance for the species was 377 m, and the maximum emigration distance was 1550 m. The analysis of patch connectivity and individual colonization revealed that the ideal distance between patches was about 300 m. Moreover, a >600 m patch distance decreased the colonization rate severely. We also observed higher immigration and emigration between patches that were clustered in close proximity. This leads us to conclude that a higher number of patches at a close distance is best suited for $P.bremeri$. We find these results to be crucial to determine a policy to protect and conserve this endangered species.

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.

인체 S100A6 단백질에 특이한 단일클론 항체

김재화,윤선영,주종혁,강호범,이영희,최용경,최인성,Kim, Jae Wha,Yoon, Sun Young,Joo, Joung-Hyuck,Kang, Ho Bum,Lee, Younghee,Choe, Yong-Kyung,Choe, In Seong 대한면역학회 2002 Immune Network Vol.2 No.3

Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.

PMA로 자극되어진 세포에서 염증 Cytokine 발현에 미치는 Bovine Lactoferrin의 생물활성 영향

정승희,강호범,김재화,윤성식,남명수,Chung, Sung-Hee,Kang, Ho-Bum,Kim, Jae-Wha,Yoon, Sung-Sik,Nam, Myoung-Soo 한국축산식품학회 2012 한국축산식품학회지 Vol.32 No.3

유청단백질의 일종인 lactoferrin은 이미 많은 보고에 의해서 여러 가지 생리활성기능이 있는 것으로 밝혀지고 있는데, U937, NK92, 수지상세포의 분화된 상태인 mutz-3와 muDC를 이용하여 과민반응과 천식, 면역 관련 유전자의 발현에 미치는 영향을 조사 한 결과 의미 있는 결과를 도출하였다. Lactoferrin 단독 또는 면역증강물질인 PMA와 혼합하여 처리 한 경우 상승효과 작용으로 과민반응과 천식, 면역 관련 유전자의 발현을 강하게 유도하였다. 이는 유청단백질의 주요 성분 중 하나인 lactoferrin이 면역기전에 중요한 역할을 하고 있다는 결과로 사료된다. Bovine lactoferrin is well known as biological activator in defense mechanism related some cells. In this study, we was investigated about the immune modulator as a role of lactoferrin through the transcriptional regulation of genes associated with hypersensitivity such as allergy, athma and inflammatory disease. Effects of inflammatory reaction of bovine lactoferrin was carried out by RT-PCR analysis from isolated total RNA treated with lactoferrin 0, 10, 50, 100, 500 ${\mu}g/mL$ and PMA 100 ng/mL. The expression of the TYROBP, PITPNA, IL-10, SLP1, DC-stamp and ICAM-1 mRNA were increased by synergy effect of bovine lactoferrin and PMA. The results of RT-PCR showed that bovine lactoferrin and PMA had an effect of immune modulator by enhancement of TYROBP, PITPNA, SLP1, DC-stamp, IL-10 and ICAM-1 gene transcription in U937, Mutz-3 and NK92 cells, respectively. Bovine lactoferrin showed a potential of biological function which could be used for industrial applications as a material of food and pharmaceutical.

Mycoplasma hominis Septic Arthritis of the Hip Developed in the Postpartum Period

Byung-Guk Kim(김병국),Hyung-Ku Youn(윤형구),Jae-Wha Kim(김재화),Hyun-Soo Ok(옥현수) 대한정형외과학회 2014 대한정형외과학회지 Vol.49 No.4

마이코플라즈마 호미니스에 의한 세균성 고관절염은 비교적 희귀할 뿐만 아니라, 출산 후 산모에게 발생했다면 이것은 매우 드문 경 우라 할 수 있다. 하지만 세균성 고관절염의 치료가 늦어지면 환자에게 매우 심각한 합병증을 유발할 수 있기 때문에 임상의는 이 질 환을 반드시 염두에 둘 필요가 있다. 이 증례는 마이코플라스마 호미니스에 의한 세균성 고관절염의 진단 및 치료에 대한 임상적 과 정을 잘 보여주고 있다고 생각하여 이에 보고하는 바이다. Septic arthritis of the hip is rarely caused by Mycoplasma hominis. It rarely develops in a patient during the postpartum period. However, delayed treatment of septic arthritis of the hip may lead to serious sequelae; therefore, it is important for clinicians not to overlook patients with the disease. This case illustrates the clinical steps in diagnosis and treatment of M. hominis septic arthritis of the hip.

대장균군 검사용 간이 시험지 개발

이인애(In-Ae Lee),김재화(Jae-Wha Kim),이회구(Hee-Gu Lee),성창근(Chang-Keun Seong),최인성(In-Seong Choe),정태화(Tai-Wha Chung) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt가 환원되어 적색 반점을 형성하는 것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 사약을 사용하여 여지에 흡착치킨 후, 건조시켜(60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No. 3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료 및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

붉은싸리버섯 추출물의 항산화 및 Human Neutrophil Elastase 저해활성

김관철 ( Kwan-chul Kim ),권용범 ( Yong-beom Kwon ),장해동 ( Hae-dong Jang ),김재화 ( Jae Wha Kim ),정재철 ( Jae Cheol Jeong ),이익수 ( Ik-soo Lee ),하병조 ( Byung-jo Ha ),유익동 ( Ick-dong Yoo ) 대한화장품학회 2016 대한화장품학회지 Vol.42 No.3

천연자원으로부터 항노화 화장품 신소재를 탐색하던 중, 국내 자생버섯의 일종인 붉은싸리버섯 자실체 추출물이 항산화 활성과 인체 호중구 엘라스타제 저해활성이 우수함을 확인하고 일련의 연구를 수행하였다. 붉은싸리버섯 추출물의 DPPH 라디칼 소거활성은 붉은싸리버섯 추출물 500 μ g/mL 처리시 117.0 mg/mL (ascorbic acid 환산값)의 매우 우수한 소거활성을 나타냈다. Peroxy 라디칼 소거활성을 oxygen radical absorbance capacity (ORAC) assay 를 통하여 측정한 결과 붉은싸리버섯 추출물 1, 10, 20 μ g/mL 처리 시, 각각 0.8, 5.2, 7.8 ORAC_Roo (trolox equivalents, 1 μ M)로 농도 의존적으로 높은 소거활성을 나타냈다. 뿐만 아니라 cellular antioxidant capacity를 DCF fluorescence intensity (% of control)로 조사한 결과에서도 붉은싸리버섯 추출물 20 μ g/mL 처리시 약 30% 이상 높은 항산화 활성을 나타냈다. Human neutrophil elastase 저해활성은 농도 의존적으로 저해활성을 나타냈으며 특히 에탄올 추출분획에서 ED₅₀ 값은 42.9 μ g/mL이었다. 붉은싸리버섯 추출물은 Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), Candida albicans (C. albicans), Aspergillus oryzae (A. oryzae) 균주 모두에서 항균활성은 나타나지 않았다. 또한 염증성 cytokine인 interleukin-10 및 interferon-γ (IFN-γ)의 생산 또는 분비 조절에는 영향을 미치지 않았다. 이상의 결과로 붉은싸리버섯 추출물은 항산화활성과 elastase 저해활성을 우수하여 피부에 자극이 없는 항노화 화장품 조성물로 유용하게 사용될 수 있음을 확인하였다. In searching for novel agents for skin anti-aging from natural resources, we found that the extract of the fruiting bodies of Ramaria formosa (R. formosa) had significant antioxidant and human neutrophil elastase (HNE) inhibitory activities. R. formosa extract exhibited a considerable DPPH radical scavenging activity with an antioxidant content of 117.0 mg/mL (ascorbic acid equivalents) at the concentration of 500 μg/mL. The capacity of R. formosa extract to scavenge peroxy radicals measured by ORAC assay also showed dose-dependent antioxidant effect with ORAC_Roo (trolox equivalents, 1 μM) values of 0.8, 5.2, and 7.8 at the concentrations of 1, 10, and 20 μg/mL. The cellular antioxidant capacity of R. formosa extract was investigated by assaying the cellular fluorescence intensity using dichlorodihydrofluorescein (DCF). The cellular oxidative stress induced by AAPH, Cu²⁺ or H2O2 in HepG2 cells was significantly attenuated by more than 30% at 20 μg/mL of R. formosa extract. HNE activity was reduced by treatment with R. formosa extract in a dose-dependent manner, and the ED₅₀ value for the ethanol extract of R. formosa was 42.9 μg/mL. R. formosa extract did not exhibited antimicrobial activity against four microorganisms including Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), Candida albicans (C. albicans), Aspergillus oryzae (A. oryzae). Furthermore, the extract did not affect the inflammatory cytokine production of interleukin-10 and interferon-γ in NK92 cells. From the above results, we found that R. formosa extract has considerable antioxidant and elastase inhibitory effects, and does not stimulate immune cells. These findings suggest that R. formosa extract may be used as a bioactive component in cosmetic composition.

젖버섯아재비 자실체로부터 분리한 Azulene계 화합물이 Interferon-γ생성에 미치는 영향

( Guang Hua Xu ),김재화 ( Jae Wha Kim ),( Gao Li ),유익동 ( Ick Dong Yoo ) 대한화장품학회 2010 대한화장품학회지 Vol.36 No.2

버섯유래 생리활성물질을 탐색하고자, 젖버섯아재비 자실체로부터 각종 컬럼크로마토그래피 및 HPLC 등 기법에 의하여 4종의 azulene계 화합물을 순수히 분리정제 하였다. 분리된 화합물은 각종 물리화학적 특성 및 분광학적 분석자료에 의하여 1-formyl-4-methyl-7-isopropyl azulene (1), lactaroviolin (2), 4-methyl-7-isopropyl-azulene-1-carboxylic acid (3) 및 1-formyl-4-methyl-7-(1-hydroxy-1-methylethyl) azulene (4)로 동정되었다. 이들 화합물의 인터페론 감마생성에 미치는 영향을 조사하였다. 화합물 1과 4는 자연살해세포주(NK92 cell)에서 인터페론 감마 생성을 농도 의존적으로 억제하였으며, 400 μM농도에서 각각 101.3 %와 92.7 %, 100 μM농도에서 각각 11.9 %와 24.1 %의 높은 저해활성을 보였으며, 화합물 2와 3은 400 μM농도에서 45.9 %와 18.0 %의 다소 낮은 저해활성을 나타내었다. Investigation of secondary bioactive metabolites from the fruiting bodies of Lactarius hatsudake led to the isolation of four azulene derivatives by means of repeated column chromatography and preparative HPLC, and they were identified as 1-formyl-4-methyl-7-isopropyl azulene (1), lactaroviolin (2), 4-methyl-7-isopropyl-azulene- 1-carboxylic acid (3), and 1-formyl-4-methyl-7-(1-hydroxy-1-methylethyl) azulene (4) by their physico-chemical properties and spectroscopic analysis. The isolated compounds were evaluated for the effects on modulation of cytokines in natural killer cell line (NK92 cells). Compounds 1 and 4 strongly inhibited IFN-γ production in a dose-dependent manner, corresponding to 101.3 % and 92.7 % inhibition at 400 μM, and 11.9 % and 24.1 % at 100 μM, respectively, whereas compounds 2 and 3 showed weak inhibitory effect on INF-γ production, corresponding to 45.9 % and 18.0 % inhibition at 400 μM.