Antitumor effect of monophosphory lipid A, polyadenylic-polyuridylic acid and cisplatin on B16 melanoma-induced pulmonary metastasis in mice

의학과/박사[한글] 종양에 대한 숙주의 방어기전은 주로 세포매개성 면역반응에 의해 유도되는 세포독성 T 림프구(cytotoxic T lymphocyte: CTL), 자연살(natural killer: NK) 세포, 림포카인 활성살(lymphokine activated killer: LAK) 세포, 종양침윤 림프구(tumor infiltrating lymphocyte: TIL) 및 대식세포의 세포독성 작용이 중요한 역활을 하는 것으로 알려져 있다. 이러한 사실에서 근래에는 이 세포들, 특히 LAK 및 TIL 세포를 interleukin(IL)-2로 시험관내에서 증식 및 활성화시켜 IL-2와 함께 종양환자에 재투여하는 자가입양 면역요법(autologous adoptive immunotherapy)이 시도되었고, 이 방법이 일부 환자에서는 임상적 치료 효과가 있음이 보고되었다. 그러나 이들 세포의 지속적인 성장과 세포독성능 유지를 위한 IL-2의 대량투여가 높은 부작용을 유발하고, 또한 몇몇 종양의 유형에만 치료효과가 있기 때문에 이 요법은 많은 문제가 제기되고 있다. Monophosphoryl lipid A(MPL)는 endotoxin인 lipopolysaccharide(LPS)의 유도체로서 독성이 매우 낮고 LPS와 매우 유사하게 면역반응을 증가시키며 특히 대식세포를 활성화시키는 동시에 interferon(IFN)-γ의 생성도 증가시키는 것으로 알려져 있다. 한편 polyadenylic-polyuridylic acid[poly(A)·poly(U))는 합성중합 핵산물질로 주로 T 세포에 작용하여IFN-γ의 생성을 증가시킴으로서 NK 세포의 세포 독성능을 증강시키는 것으로 보고되고 있다. 이에 본 연구에서는 MPL, poly(A)· Poly(U) 및 항암제인 cisplatin을 단독 혹은 병용투여하여 마우스 악성 흑색종의 실험적 폐전이에 미치는 항종양효과를 관찰하고 그 작용기전을 규명하고자 하였다. 1. MPL, poly(A) · poly(U) 및 cisplatin 투여군은 대조군에 비해 전이성 폐암 형성이 현저히 억제되었다. 2. Cisplatin과 MPL 또는 Poly(A)·Poly(U)의 병용투여는 병용효과가 없었으나, MPL 및 Poly(A) · poly(U)에 의한 항암효과는 단독투여보다 병용투여가 증가하는 경향을 보였다. 3. 생체내 MPL 및 Poly(A)· poly(U)의 투여시 비장 림프구의 세포독성능은 YAC-1 및 악성 흑색종세포에 대해 모두 증가하는데 반하여, 대식세포의 세포독성능은 YAC-1에 대해서만 증가하는 것으로 나타났다. 그리고 그 효과는 MPL 투여군이 Poly(A)· Poly(U) 투여 군보다 더 높은 것으로 나타났다. 한편 시험관내에서 MPL 및 poly(A)· Poly(U)의 자극은 비장 림프구 및 대식세포의 세포독성능을 증가시키지 못하였다. 4. 리장 림프구의 IL-2 및 IFN-γ의 생성은 MPL 및 po1y(A)· poly(U) 투여군에서 대조군에 비해 현저히 증가하였으며, MPL 투여군이 poly(A)·poly(U) 투여군보다 높은 증가를 보였다. 이들 세포의 IL-4의 생성은 poly(A)· poly(U) 투여군과 대조군사이에는 차이가 없었으며, MPL 투여군에서는 오히려 현저히 감소하였다. 이상의 결과를 종합하여 볼때 마우스에 있어서 MPL 및 poly(A)· poly(U) 투여는 악성 흑색종의 실험적 폐전이를 억제하는 것으로 나타났으며, 이들의 억제현상은 림프구의 IL-2 및IFN-γ의 생성을 증가시켜, NK 세포 및 대식세포의 세포독성능의 증가에 의해 기인된 것으로 생각된다. 특히 MPL이 poly(A)· poly(U)보다 높은 암 억제 효과를 나타내는 경향을 보이며, 세포 매개성 면역기능에 관여하는 IL-2 및 IFN-γ의 생성을 현저하게 증가시키는 점을 고려할때, 향후 유용한 면역치료제로 쓰일 수 있을 것으로 사료된다. Antltumor effect of monophosphoryl lipid A, polyadenylic-polyuridylic acid and cisplatin on B16 melanoma-induced pulmonary metastasis in mice Chul Ho Cho Department of Medical science The Graduate School, Yonsei University (Directed by Professor Joo Deuk Kim) Despite aggressive surgery, radiation and combined chemotherapy, recurrence after treatment of primary cancer is common. Complete eradication of cancer cells with these modalities is hard, if at all, to achieve. Immunity is usually depressed in hosts with tumor and a strong relationship exists between immunocompetence and prognosis of cancer. Immunotherapy thus aims at stimulating natural host defense mechanism to facilitate tumor regression. Augmentation of natural killer(NK) cell cytotoxicity, macrophage activation, and interferon (IFN)-γ-mediated T Iymphocyte proliferation are believed to be the main antitumor effects of immunotherapy. Monophosphoryl lipid A(MPL), a derivative of lipopolysaccharide(LPS), which has antitumor activity with low toxicity, has been shown to enhance antibody production with increased IFN-γ in both young adult and aging mice. Polyadenylic-polyuridrlic acid[poly(A) · poly(U)] is a nontoxic and metabolically stable immunomodulator, capable of efficiently stimulating various compartments of host-immune system, thereby enhancing their antitumor activity. It has been successfully used in adjuvant treatment of human breast and stomach cancer, increasing patient survival and decreasing recurrence. This study looted into antitumor effects of MPL, poly(A) · poly(U) and cisplatin, used alone or in combination, against Bl6 melanoma lung metastasis in mice, and the following results were obtained: 1) Antitumor effects of MPL, poly(A) · poly(U) and cisplatin were significantly higher in the mice treated with these agents than the saline-treated controls. 2) The combined use of MPL and poly(A) · poly(U) tended to show, though not significant, higher antitumor effect than the sole use of either MPL or poly(A) · poly(U) in mice not previously treated with cisplatin, but not in mice pretreated with cisplatin. 3) In vivo inoculation of MPL and poly(A) · poly(U) significantly enhanced cytotoxicity of spleen cells against both YAC-1 cells and Bl6 melanoma cells, but enhanced cytotoxicity of peritoneal macrophages only against the former(YAC-1 cells). In vitro, it failed to increase cytotoxicity of either spleen cells or peritoneal macrophages. 4) In vivo IL-2 and IFN-γ Production were significantly higher in the mice treated with MPL or poly(A)·poly(U) than the controls with the production in the mice treated with MPL higher than with poly(A)·poly(U). IL-4 production in vivo was not different between the mice treated with poly(A)·poly(U) and the controls, but in the MPL-treated mice, it was significantly lower than in the controls. Antitumor activity of MPL and Poly(A)·Poly(U) against metastatic lung cancer appears to arise from increased cytotoxicy of NK cells and macrophages, mediated by the increased IL-2 and IFN-γ production. In Particular, compared with poly(A)·poly(B), MPL, the substance showing as in this study a high antitumor effect and significantly increased production of IL-2 and IFN-γ, the cytokines believed to be important in cell-mediated immunity, appears premising as a potentially useful immunotherapeutic agent. [영문] Despite aggressive surgery, radiation and combined chemotherapy, recurrence after treatment of primary cancer is common. Complete eradication of cancer cells with these modalities is hard, if at all, to achieve. Immunity is usually depressed in hosts with tumor and a strong relationship exists between immunocompetence and prognosis of cancer. Immunotherapy thus aims at stimulating natural host defense mechanism to facilitate tumor regression. Augmentation of natural killer(NK) cell cytotoxicity, macrophage activation, and interferon (IFN)-γ-mediated T Iymphocyte proliferation are believed to be the main antitumor effects of immunotherapy. Monophosphoryl lipid A(MPL), a derivative of lipopolysaccharide(LPS), which has antitumor activity with low toxicity, has been shown to enhance antibody production with increased IFN-γ in both young adult and aging mice. Polyadenylic-polyuridrlic acid[poly(A) · poly(U)] is a nontoxic and metabolically stable immunomodulator, capable of efficiently stimulating various compartments of host-immune system, thereby enhancing their antitumor activity. It has been successfully used in adjuvant treatment of human breast and stomach cancer, increasing patient survival and decreasing recurrence. This study looted into antitumor effects of MPL, poly(A) · poly(U) and cisplatin, used alone or in combination, against Bl6 melanoma lung metastasis in mice, and the following results were obtained: 1) Antitumor effects of MPL, poly(A) · poly(U) and cisplatin were significantly higher in the mice treated with these agents than the saline-treated controls. 2) The combined use of MPL and poly(A) · poly(U) tended to show, though not significant, higher antitumor effect than the sole use of either MPL or poly(A) · poly(U) in mice not previously treated with cisplatin, but not in mice pretreated with cisplatin. 3) In vivo inoculation of MPL and poly(A) · poly(U) significantly enhanced cytotoxicity of spleen cells against both YAC-1 cells and Bl6 melanoma cells, but enhanced cytotoxicity of peritoneal macrophages only against the former(YAC-1 cells). In vitro, it failed to increase cytotoxicity of either spleen cells or peritoneal macrophages. 4) In vivo IL-2 and IFN-γ Production were significantly higher in the mice treated with MPL or poly(A)·poly(U) than the controls with the production in the mice treated with MPL higher than with poly(A)·poly(U). IL-4 production in vivo was not different between the mice treated with poly(A)·poly(U) and the controls, but in the MPL-treated mice, it was significantly lower than in the controls. Antitumor activity of MPL and Poly(A)·Poly(U) against metastatic lung cancer appears to arise from increased cytotoxicy of NK cells and macrophages, mediated by the increased IL-2 and IFN-γ production. In Particular, compared with poly(A)·poly(B), MPL, the substance showing as in this study a high antitumor effect and significantly increased production of IL-2 and IFN-γ, the cytokines believed to be important in cell-mediated immunity, appears premising as a potentially useful immunotherapeutic agent.restrictio

Diagnostic value of carcinoembryonic antigen (CEA) in ascitic fluid

의학과/석사[한글] 복수는 여러 가지 양성 및 악성질환에 의해서 발생되는데 복수환자에 있어서 복수가 양성질환에 의한 복수인지 악성질환에 의한 복수인지 악성질환에 의한 복수인지 감별하기 어려운 경우가 있고, 이의 감별을 위해 여려가지 방법이 이용되고 있다. Carcinoembryoni c antigen (이하 CEA로 약함)은 흉막액내의 농도를 측정함으로써 흉곽내 악성질환 존재여부를 아는데 도움이 된다는 보고들이 있었고, 또한 Loewenstein등(1978)과 Pare등(1983) 은 암환자의 복수 CEA치가 혈청 CEA치보다 현저히 높았음을 보고하였다. 저자는 복수내 CEA가 복강내 악성질활의 존재여부와 악성질환의 복막전이 여부를 아는데 도움이 되는지를 알아보기 위하여 1985년 3월부터 1985년 10월까지 연세대학교 의과대학 부속 세브란스병원 내과에 입원하였던 복수한자 75예를 대상으로 복수 및 혈청 CEA치 를 측정하고 이를 비교 분석하여 다음과 같은 결과를얻었다. 1) 양성복수군의 복수 및 혈청 CEA치의 평균치는 각각 2.46ng/ml, 3.83ng/ml이었으며, 복수의 악성 및 양성 감별을 위한 기준치는 양성복수군외 복수 및 혈청 CEA치의 평균치에 표준편차 2배를 합한 간인 8,72ng/ml, 9.95ng/ml로 정하였다. 2) 복수 CEA치가 기준치 (8.72ng/ml) 이상은 악성복수군 9예중 6예(66.7%), 악성질환군 36예중 14예(38.9%), 양성복수군 30예중 1예 (3.3%)이었다. 복수 CEA치가 57ng/ml 이상은 모두 16예이었는데 악성복수군이 6예, 악성질환이 있었다. 3) 혈청 CEA치가 기준치 (9.95ng/ml) 이상은 악성복수군 9예중 5예(55.5%), 악성질환군 36예중 12예 (33.3%), 양성복수군 30예중 2예(6.7%)이었다. 혈청 CEA치가 50ng/ml 이상은 모두 8예이었는데 악성복수군이 3예, 악성질환군이 5예로 8예 전 예에서 복강내 악성질환이 있었다. 4) 복수 CEA치는 기준치 이상이고 혈청 CEA치가 기준치 이하는 5예이며, 이중 악성복수군이 1예, 악성질환군은 3예로 4예(80%)에 복강내 악성질환이 있었다. 5) 복강내 악성질환의 진단에 있어서 복수 CEA검사의 96.7%, 양성(陽性) 예측도는 95.2%로 매우 높았고, 감수성은 44.4%, 음성(陰性) 예측도는 53.7%로 낮았다. 6) 복강내 악성질환의 진단에 있어서 혈청 CEA검사의 특이성은 93.3%, 양성 예측도는 89.5%로 높았고, 감수성은 37.8%, 음성 예측도는 50.0%로 역시 낮았으며, 복수 CEA검사보다 낮은 진단율을 보였다. 이상의 결과로 복수 CEA검사는 감수성과 음성 예측도는 낮으나 특이성과 양성 예측도가 높아 복강내 악성질환의 존재여부를 아는데 도움이 되며, 혈청 CEA검사보다 진단율이 높았다. 또한 복수 CEA치가 50ng/㎖이상인 경우는 악성질환의 복막전이 여부를 아는데 도움 이 될 것으로 사료된다. [영문] Ascites may be caused by various benign and malignant diseases but it is sometimes difficult to determine whether the ascites is caused by benign diseases or malignant diseases. Various diagnostic methods such as ascites level and ascites/serum ratio of protein and LDH, cytology and peritoneoscopy have been used in differentiating between benign and malignant ascites. Carcinoembryonic antigen(CEA) level in pleural effusion is known to be useful to make the diagnosis of the malignant diseases in thoracic cavity. In order to determine whether ascites CEA is of help to make the diagnosis of malignant diseases in abdominal cavity and their peritoneal metastasis, both ascites CEA were measured and analyzed in 75 patients with ascites who were admitted to Yonsei University Severance Hospital from March 1985 to October 1985. The results were as follows : 1. The mean value of CEA level in ascites and serum in benign ascites group was 2.46ng/ml, and 3.83n/ml respectively. The upper limit(mean+2 S.D.)of ascites and serum CEA in benign ascites group was 8.72ng/ml, and 9.95ng/ml respectively. 2. Ascites CEA level was higher than 8.72ng/㎖ in 6 of 9 patients with malignant ascites(66.7%), 14 of 36patients with malignant diseases(38.9%), and one of 30 patients with benign ascites(3.3%). In 16 patients ascites CEA lev디 was higher than 50ng/㎖(6 patients with malignant ascites, and 10 patients with malignant diseases). All of them had malignant diseases in abdominal cavity. 3. Serum CEA level was higher than 9.95ng㎖ in 5 of 9 patients with malignant ascites(55.5%), 12 of 36 patients with malignant diseases(33.3%), and 2 of 30 patients with benign ascites(6.7%), In 8 patients serum CEA level was higher than 50ng/㎖(3 patients with malignant ascites and 5 patients with malignant diseases). All of them had malignant diseases in abdominal cavity. 4. In 5 patients ascites CEA level was higher than 8.72ng/㎖ but serum CEA level was lower than 9.95ng/㎖. Four had malignant diseases in abdominal cavity(one patient with malignant ascites and 3 patients with malignant diseases). 5. The specificity of ascites CEA(8.72ng/㎖)for malignant diseases in abdominal cavity was 96.7%, the positive predictability 95.2%, the sensitivity 44.4%, and the negative predictability 53.7%. 6. The specificity of serm CEA(9.95ng/㎖)for malignant diseases in abdominal cavity was 93.3%, the positive predictability 89.5%, the sensitivity 37.8%, and the negative predictability 50.0%, which were slightly lower than those of ascites CEA. In summary, the measurement of ascites CEA level is useful in determining the existence of malignant diseases in abdominal cavity and higher than that of serum CEA in the diagnostic accuracy. Also it is useful in predicting the peritoneal metastasis in cases with ascites CEA level higher than 50ng/㎖.restrictio