SSX基因家族HLA-A2.1限制性CTL表位预测及其MHC-Ⅰ亲和力分析  

Prediction of HLA-A2.1 restricted CTL epitope of SSX gene family and affinity assay of these epitopes with MHC-Ⅰ molecule

作　　者：何仰东[1] 吴玉章[1] 林治华[2] 贾正才[1] 姜曼[1] 黎万玲[1] 付晓岚[1] 邓一静[1] 倪兵[1] 

机构地区：[1]第三军医大学基础医学部全军免疫学研究所,重庆400038 [2]重庆工学院生物工程学院,重庆400050

出　　处：《第三军医大学学报》2006年第9期872-875,共4页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目("973"项目)(2003CB514108);国家自然科学基金资助项目(30571835)~~

摘　　要：目的 通过对肿瘤基因SSX家族不同成员进行CTL表位预测，并对其与MHC-I亲和力进行分析，为探索基于SSX肿瘤基因家族的免疫治疗奠定基础。方法 利用基于超基序结合量化基序方案开发的HLA-A2．1限制性CTL表位预测软件，对SSX基因家族不同成员进行CTL表位预测，然后合成SSX相关待测表位肽P1（AMTKLGFKA）、P4（AMT-KLGFNV）、P5（AMTKLGFKV）和P6（VMTKLGFKV），再对这些合成肽与MHC-I结合亲和力及其结合物稳定性进行实验分析。结果与实验阳性肽（HBcAg，18～27）相比，除P1外，P4、P5、P6均显示较高的结合亲和力；而结合稳定性实验（DC50）结果显示，P1、P4与阳性对照肽（HBcAg，18～27）DC。均大于8h，而P5和P6的DC50介于2～4h之间。结论 计算机软件预测的CTL表位肽，经过体外亲和力与结合稳定性实验筛选到2条理想的表位肽，为该表位的下一步体内鉴定奠定基础。Objective To explore the immunotherapeutic strategy based on SSX tumor gene family by the CTL epitope prediction of the members of SSX tumor gene family and affinity assay of the epitope combining with MHC-Ⅰmolecule. Methods The CTL epitope in the members of SSX gene family were predicted with the predictive software of HLA-A2.1 restricted CTL epitope. The 4 predicted epitope peptides were synthesized ： P1 （AMTKLGFKA）, P4 （AMTKLGFNV）, P5 （AMTKLGFKV） and P6 （VMTKLGFKV）. After the affinity between these peptides and MHC-Ⅰ molecule were examined respectively, the combining stability between these peptides and the MHC- Ⅰ molecule were investigated respectively. Results As compared with positive control peptide HBcAg18 -27, all of the synthesized peptides except P1 showed high affinity with MHC- Ⅰ molecule. The combining stability assay indicated that dissociation complex 50 （ DC50 ） for P1, P4 and the positive control HBcAg 18 -27 was all above 8.0 h, but DC50 for P5 and P6 was between 2 h and 4 h. Conclusion Two ideal peptides were screened out from the predicted CTL epitope peptides by in vitro MHC- Ⅰ combining affinity and stability assay, which provided the experimental data for the further identification of these epitope peptides in vivo.

关 键 词：SSX肿瘤基因 表位预测 HLA-A2.1限制性 结合亲和力 

分 类 号：R392.33[医药卫生—免疫学] R392.3[医药卫生—基础医学]