CAJ | 학술논문

目的：研究抗 CTLA-4（cytotoxic T lymphocyte antigen-4，CTLA-4）抗体（9H10）对小鼠 Lewis 肺癌的抑瘤作用及其机制。方法选取健康适龄 C57小鼠，皮下接种小鼠 Lewis 肺癌细胞系，建立皮下移植瘤模型。12只荷瘤小鼠分为2组，分别为空白对照组和实验组，分别腹腔注射给予生理盐水和 CTLA-4抗体（9H10），通过肿瘤体积、重量及肿瘤生长抑制率评估9H10的抑瘤作用；免疫组织化学染色检测脾组织 T 细胞亚型（CD4及 CD8阳性 T 细胞）及肿瘤微血管密度（MVD），通过组间对照分析抗 CTLA-4对肿瘤微环境免疫细胞及血管生成的影响。结果CTLA-4抗体可延缓荷瘤小鼠的肿瘤生长速度，延长其生存时间。免疫组织化学染色检测荷瘤小鼠脾脏中 CD4及 CD8阳性 T细胞。结果显示，实验组与对照组 CD4阳性 T 细胞的数量分别为（49．05±7．31）％和（23．45±8．06）％，与对照组比较，应用抗 CTLA-4处理后 CD4表达阳性率明显增高，具有统计学意义（P ＜0．01）。实验组和对照组中 CD8表达的阳性率分别为17．25±2．86和11．32±0．71，与对照组比较，应用抗 CTLA-4处理后 CD8表达的阳性率明显增高，具有统计学意义（P ＜0．05）。用抗 CD34单克隆抗体标记肿瘤微血管内皮细胞，计数肿瘤间质 MVD。实验组和对照组的 MVD 值分别为37．20±2．84和75．72±14．49，CTLA-4使肿瘤 MVD 明显减少，具有统计学意义（P ＜0．001）。结论CTLA-4抗体对荷瘤小鼠 Lewis 肺癌具有抑制作用，荷瘤小鼠脾组织中 CD4及 CD8表达均增高，肿瘤间质的MVD 值显著降低。CTLA-4在提供机体抗肿瘤免疫及抑制肿瘤血管生成中均发挥重要作用。
목적：연구항 CTLA-4（cytotoxic T lymphocyte antigen-4，CTLA-4）항체（9H10）대소서 Lewis 폐암적억류작용급기궤제。방법선취건강괄령 C57소서，피하접충소서 Lewis 폐암세포계，건립피하이식류모형。12지하류소서분위2조，분별위공백대조조화실험조，분별복강주사급여생리염수화 CTLA-4항체（9H10），통과종류체적、중량급종류생장억제솔평고9H10적억류작용；면역조직화학염색검측비조직 T 세포아형（CD4급 CD8양성 T 세포）급종류미혈관밀도（MVD），통과조간대조분석항 CTLA-4대종류미배경면역세포급혈관생성적영향。결과CTLA-4항체가연완하류소서적종류생장속도，연장기생존시간。면역조직화학염색검측하류소서비장중 CD4급 CD8양성 T세포。결과현시，실험조여대조조 CD4양성 T 세포적수량분별위（49．05±7．31）％화（23．45±8．06）％，여대조조비교，응용항 CTLA-4처리후 CD4표체양성솔명현증고，구유통계학의의（P ＜0．01）。실험조화대조조중 CD8표체적양성솔분별위17．25±2．86화11．32±0．71，여대조조비교，응용항 CTLA-4처리후 CD8표체적양성솔명현증고，구유통계학의의（P ＜0．05）。용항 CD34단극륭항체표기종류미혈관내피세포，계수종류간질 MVD。실험조화대조조적 MVD 치분별위37．20±2．84화75．72±14．49，CTLA-4사종류 MVD 명현감소，구유통계학의의（P ＜0．001）。결론CTLA-4항체대하류소서 Lewis 폐암구유억제작용，하류소서비조직중 CD4급 CD8표 체 균 증 고，종류간질적MVD 치현저강저。CTLA-4재제공궤체항종류면역급억제종류혈관생성중균발휘중요작용。
Objective To investigate the antitumor effects of cytotoxic T lymphocyte antigen-4 antiboby(9H10)on mice Lewis lung cancer.Methods The transplantation tumor models were established by injection of Lewis lung cancer cells subcutaneously to C57BL/6 mice.Twelve mice were randomly divided into two groups:The 9H10 therapy group with intraperitoneal injection of cytotoxic T lymphocyte antigen-4 antibody and the control group with the injection of 0.9% sodium chloride respectively.The antitumor effects were investigated by measuring tumor volume,the dynamic change of tumor weight and the inhibition rate of tumor growth.The expression ratio of CD4 and CD8 positive T cells in spleen were detected by immunohistochemistry.Microvessel density (MVD)was assessed by CD31 immunostaining. Results The growth rate of tumor bearing mice were delayed and the survival time were prolonged.The CD4 positive T cells in the spleen of tumor bearing mice were 49.05±7.31% and 23.45±8.06% in 9H10 group and control group respectively.The positive rate of CD4 T cells were significantly increased comparing with control group after CTLA-4 antibody treatment (P <0.01).The CD8 positive T cells were 17.25±2.86 and 11.32±0.71 in 9H10 group and con-trol group respectively.The positive rate of CD8 T cells were significantly increased in the experimental group than in the control group(P <0.05).The tumor interstitial microvessel density (MVD)were 37.20±2.84 和 75.72±14.49 in the experimental group and control group respectively.The MVD significantly decreased after CTLA-4 antibody treat-ment (P <0.01).Conclusion The growth of Lewis lung cancer were inhibited after CTLA-4 antibody treatment.The expression ratio of CD4 and CD8 positive T cells were both increased and the MVD value was lowered in the spleen of Lewis cancer bearing mice after CTLA-4 antibody treatment.CTLA-4 may play significant roles both on increasing the tumor immune and inhibiting tumor angiogenesis.