目的: 研究粒细胞巨噬细胞集落刺激因子（granulocytemacrophage colony stimulating factor, GMCSF)膜修饰小鼠黑素瘤B16.F10细胞制备的疫苗对小鼠种植肿瘤的抑制作用。方法：采用生物素链亲合素GMCSF融合蛋白技术制备GMCSF膜修饰B16.F10黑素瘤细胞疫苗，将BALB/c小鼠随机分为GMCSF膜修饰B16.F10细胞疫苗组、GMCSF与B16.F10细胞混合疫苗组、B16.F10细胞疫苗组、GMCSF组、生理盐水组。各组于第1天和第7天分别进行免疫接种，第14天皮下注射0.2 ml B16.F10细胞（1×106个细胞）进行攻击，观察各组小鼠无瘤率和生存期；攻击后24 d 用ELISA法检测小鼠脾细胞INFγ分泌水平。结果：接受B16.F10细胞攻击后第60天和第90天，GMCSF膜修饰B16.F10细胞疫苗组小鼠均未成瘤,存活率为100%；GMCSF与 B16.F10细胞混合组无瘤率分别50%和40%，存活率分别为70%和40%；B16.F10细胞疫苗组无瘤率均为20%，存活率分别为80%和20%；GMCSF组和生理盐水组无瘤率为0，无小鼠存活。GMCSF膜修饰B16.F10细胞疫苗组的IFNγ分泌量比其他组明显升高（P<0.01）。结论：GMCSF膜修饰B16.F10肿瘤细胞疫苗可以激发BALB/c小鼠特异性抗肿瘤免疫反应，有效防止B16.F10肿瘤细胞的攻击。

Objective:To study the inhibitory effect of GMCSF（granulocytemacrophage colony stimulating factor) membraneanchored B16.F10 (murine melanoma cell line) vaccine against implanted tumors in mice. Methods: GMCSF membraneanchored B16.F10 vaccine was prepared by fusing biotinavidinGMCSF together. BALB/c mice were randomly divided into 5 groups: GMCSF membraneanchored B16.F10, B16.F10 mixed with GMCSF, B16.F10 cells, GMCSF and 0.9% sodium chloride. At day 1 and 7, all BALB/c mice were immunized; at day 14, all the mice received subcutaneous injection of 0.2 ml wildtype B16.F10 cells (1×106 cells). Tumorfree rate and survival time of different groups were measured. INFγ levels in splenocytes were detected by ELISA at day 24 in different groups.Results: Sixty days and 90 d after subcutaneous injection with B16.F10 cells, tumorfree rate and survival rate of mice in GMCSF membraneanchored B16.F10 group, B16.F10 mixed with GMCSF group, B16.F10 cell group were 0% and 0%, 100% and 100%; 50% and 40%, 70% and 40%; 20% and 20%, and 80% and 20%, respectively. The tumorfree rate and survival rate of mice in GMCSF group and 0.9% sodium chloride group were both 100% and 0%, respectively. INFγ levels in splenocytes in GMCSF membraneanchored B16.F10 group were significantly higher than those of other groups（P<0.01）. Conclusion: GMCSF membraneanchored B16.F10 vaccine can induce specific antitumor immunity in mice and can prevent later B16.F10 tumor cells challenge.

广东省自然科学基金重点资助项目(No.06104396)