To construct cDNA library of nasopharyngeal carcinoma （NPC） and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which...To construct cDNA library of nasopharyngeal carcinoma （NPC） and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX （serological identification of antigen by recombinant cDNA expression library）. Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64 × 10^6 recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31, SI00 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.展开更多
Identification of hepatocellular carcinoma- associated tumor antigens is necessary and pivotal for specific immunotherapy in hepatocellular carcinoma (HCC) patients. In the present study, HCC cDNAs are constructed int...Identification of hepatocellular carcinoma- associated tumor antigens is necessary and pivotal for specific immunotherapy in hepatocellular carcinoma (HCC) patients. In the present study, HCC cDNAs are constructed into ZAP cDNA expression library and screened by sera of patients with HCC. The positive clones are DNA sequenced and analyzed by bioinformatics. Thirty-one genes of hepatocellular carcinoma-associated tumor antigens are identified, of which 1 is unknown and 30 are known. The proteins encoded by these known genes can be classified into 8 categories: constitutive molecules of hepatocytes, RNA transcription and splicing-associated molecules, protein metabolism-associated molecules, energy synthesis-associated molecules, signal transduction molecules, cell adhesion molecules, immunosuppressive molecules, and proteins with unknown function. Among these genes, CAGE is a cancer-testis (CT) antigen. It is concluded that identification of hepatocellular carcinoma-associated tumor antigens provides potential targets for immunotherapy of HCC patients and facilitates explanation of carcinogenesis of HCC.展开更多
文摘目的:通过重组cDNA文库的血清学筛选(SEREX)技术获得来自非洲爪蟾(Xenopus laevis)肝脏肿瘤文库中具有潜在治疗价值的异种同源抗原的cDNA序列。方法:用人肝癌细胞株SMMC-7721免疫家兔后收集的抗血清作为筛选血清,采用SEREX技术筛选Xenopuslaevis肝脏肿瘤cDNA表达文库。结果:通过两轮筛选获得19个阳性克隆,通过DNA序列测定和分子生物学技术确定这19个克隆编码16个蛋白。其中Transgelin 2、RASguanylreleasing protein1和Chromobox protein homolog6可能与肝细胞肝癌有关。结论:用异种免疫血清筛选异种同源肿瘤抗原拓宽了SEREX技术的应用范围,鉴定Transgelin2、RASguanyl releasing protein1和Chromobox protein homolog6可能为异种同源抗原。
文摘To construct cDNA library of nasopharyngeal carcinoma （NPC） and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX （serological identification of antigen by recombinant cDNA expression library）. Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64 × 10^6 recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31, SI00 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.
文摘Identification of hepatocellular carcinoma- associated tumor antigens is necessary and pivotal for specific immunotherapy in hepatocellular carcinoma (HCC) patients. In the present study, HCC cDNAs are constructed into ZAP cDNA expression library and screened by sera of patients with HCC. The positive clones are DNA sequenced and analyzed by bioinformatics. Thirty-one genes of hepatocellular carcinoma-associated tumor antigens are identified, of which 1 is unknown and 30 are known. The proteins encoded by these known genes can be classified into 8 categories: constitutive molecules of hepatocytes, RNA transcription and splicing-associated molecules, protein metabolism-associated molecules, energy synthesis-associated molecules, signal transduction molecules, cell adhesion molecules, immunosuppressive molecules, and proteins with unknown function. Among these genes, CAGE is a cancer-testis (CT) antigen. It is concluded that identification of hepatocellular carcinoma-associated tumor antigens provides potential targets for immunotherapy of HCC patients and facilitates explanation of carcinogenesis of HCC.
