GAGE-1 DNA肿瘤疫苗的构建及其抗肿瘤治疗效果的试验研究

Construction of the GAGE-1 DNA tumor vaccine and its therapeutic efficacy to the tumor

核酸疫苗的接种有多种途径,包括肌内、皮内、皮下注射途径、基因枪免疫、黏膜免疫途径等.不同免疫途径的免疫机制不同,所诱导的免疫保护力的强弱和维持的时间也不尽相同.GAGE基因通常表达于睾丸组织和部分恶性肿瘤组织中,被认为可能是理想的癌症诊断的标记和治疗的靶位.GAGE-1这个基因就属于这个家族.在本研究中,将C57 BL/6小鼠随机分为4组,分别接种实验组或对照组各三次.末次免疫后10d小鼠用于肿瘤细胞攻击试验:分别于左背部、右背部皮下种植B16肿瘤细胞、B16/GAGE-1肿瘤细胞.种植肿瘤细胞(荷瘤)后观察成瘤时间、肿瘤大小,并对小鼠脾细胞进行CTL细胞杀伤活性实验来观察DNA疫苗引起的免疫反应.

There are many routes of DNA vaccine immunization, including intramuscularly injection, in- tradermal injection, subcutaneous injection, gene gun immunization, mucosa immunization, and so on. Different inoculation routes could result in different immunologic potency and persistent time. GAGE gene family are known as the ideally marker and target for diagnosis and therapy of cancer. GAGE-1 gene is a member of this family. In this research, C57BL/6 mice were divided into 4 groups, subcutaneously injected with pcDNA3.1 ＋/GAGE-1 （experimental group 1）, subcutaneously injected with pcDNA3.1＋/GAGE-1/ IL-2 （experimental group 2）, pcDNA3.1＋ （control group 1）, pcDNA3.1＋/IL-2 （control group 2）, which were immunized thrice at week 0,2,4 respectively. 10d after the last boost all mice of each group were subcutaneously inoculated with B16 cells on the left back while with B16- GAGE-1 cells on the right back. The time of visible tumor formation and the size of tumor were recorded and observed. The life span and survival rate of mice after loaded with the tumor ceils were analyzed by Log2-Rank statistics and the CTL values.