Zhang Yeping, Zhu Ping, Shi Yongjin, Liu Jihua, Pu Dingfang, Cao Xianghong, Zhu Qiang, Wang Yijia, Ma Mingxin, Yu Jiren
Department of Hematology, Beijing Hospital, Beijing 100730, China.
Zhonghua Xue Ye Xue Za Zhi. 2002 Feb;23(2):68-72.
To investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.
The specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.
TCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.
The idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.
研究含不同抗原决定簇的TCR独特型DNA疫苗对BALB/c小鼠体内人CEM淋巴瘤细胞的杀伤活性。
采用RT-PCR技术获取CEM细胞系编码TCRVβ区的特异性重排基因片段。将PCR产物克隆至真核表达载体pcDNA3，以此作为DNA疫苗及PCR扩增不同抗原决定簇的模板。分别扩增编码不同抗原决定簇的基因片段并克隆至pcDNA3。通过肌肉注射DNA疫苗免疫实验小鼠。采用间接免疫荧光法检测特异性抗独特型抗体。
CEM细胞系的TCRβV含有5个抗原决定簇。用含全部5个决定簇的DNA疫苗免疫的6只小鼠均检测到特异性抗独特型抗体（最高效价为1:480）。在用含5个抗原决定簇中4个的DNA疫苗免疫的6只小鼠中，虽有4只检测到抗体，但抗体效价较低（最高效价为1:80）。含5个决定簇中2个的DNA疫苗不能诱导产生特异性抗体。
含完整TCRβV 5个抗原决定簇的独特型DNA疫苗可在BALB/c小鼠体内诱导产生特异性抗淋巴瘤独特型抗体。
