本文選題：SSX + HLA-A2限制性��； 參考：《第三軍醫(yī)大學》2006年碩士論文

【摘要】： 基于抗原表位肽的疫苗,由于其中表位肽的化學成分相對簡單、穩(wěn)定、容易構(gòu)建、可以避免各種污染及潛在的致癌物質(zhì)等方面的優(yōu)勢,已經(jīng)成為腫瘤免疫治療研究的主要策略之一。目前這種疫苗形式中的抗原表位肽主要是來源于腫瘤/睪丸抗原(Cancer/testis antigen,CT抗原)。SSX基因家族是CT抗原之一,與其它多成員組成的CT抗原家族一樣,它由9個不同成員組成。在某一種腫瘤細胞中不會同時表達所有SSX家族成員,而是僅表達其中一種或幾種。如對6種不同組織類型的10個滑膜肉瘤細胞系,進行SSX-1、SSX-2、SSX-3、SSX-4、SSX-5的mRNA表達調(diào)查發(fā)現(xiàn),其表達陽性率分別為90%、50%、10%、60%和50%,兩種以上基因同時表達的達60%,全部此類腫瘤細胞至少表達一種上述基因。而對26例原發(fā)性肝癌(hepatocellular carcinoma,HCC)組織進行SSX-1、SSX-2和SSX-5的mRNA表達分析,其表達陽性率分別為61.5%、34.6%和46.2%,兩種以上基因同時表達的有4例(15.4%),至少有一種基因表達的有17例(65.4%)。因而針對其中某一種成員的抗原表位肽腫瘤疫苗的實際應用將會存在一定的局限性�；诖�,研究開發(fā)能同時針對整個SSX家族所有成員的表位肽疫苗將會有廣闊的應用前景。 本課題將SSX家族的9個成員分別輸入林治華教授基于超基序結(jié)合量化基序列開發(fā)的CTL表位預測軟件,并結(jié)合著名的美國國立衛(wèi)生研究所的BIMAS軟件(http://www-bimas.cit.gov/ molbio/hla_bind/)和德國海德堡生物醫(yī)學信息中心根據(jù)特定肽的錨定殘基和輔助殘基位置的氨基酸類型來評分的SYFPEITHI軟件(http://www.syfpeithi.de/),分析它們的預測結(jié)果。發(fā)現(xiàn)SSX57-65是所有成員中預測分值相對較高的肽片段,并且它們之間的序列相似程度非常高,僅在第1、8、9位上有所區(qū)別,而其它位點序列完全一致。根據(jù)這些結(jié)果,我們合成了可以代表SSX57-65全部成員的4條肽,即(1) P1: AMTKLGFKA (SSX-2, SSX-3, SSX-5, SSX-7, SSX-9), (2) P4: AMTKLGFNV (SSX-6, SSX-8), (3) P5: AMTKLGFKV (SSX-1), (4) P6: VMTKLGFKV (SSX-4)。在分析此4條預測表位肽與MHC-I分子結(jié)合的親和力大小和結(jié)合穩(wěn)定性的基礎上,我們再用它們在體外刺激健康人PBMC,用可以評價特異性CTL的
[Abstract]:Because the epitope peptide based vaccine is relatively simple, stable and easy to construct, it can avoid all kinds of pollution and potential carcinogens. It has become one of the main strategies in tumor immunotherapy. At present, the epitope peptide of this vaccine is mainly derived from tumor / testis antigen Cancer-testis antigen.SSX gene family is one of CT antigens. In a tumor cell, all members of SSX family are not expressed at the same time, but only one or several of them are expressed. For 10 synovial sarcoma cell lines of 6 different tissue types, the mRNA expression of SSX-1pSSX-2mSSX-3nSSX-3nSX-4nSSX-5 was investigated. The positive rates of SSX-4SSX-5 expression were 90% and 50101060% and 50%, respectively. The two or more genes were expressed at the same time, and all of these tumor cells expressed at least one of the above genes. The positive rates of SSX-1mSSX-2 and SSX-5 were 61.5% and 46.2%, respectively. The positive rates of SSX-1SSX-2 and SSX-5 were 61.5% and 46.2%, respectively. There were 4 cases with two or more genes expressed at the same time and 17 cases with at least one gene expression (65.4%). Therefore, the application of epitope peptide tumor vaccine against one of its members will be limited. Therefore, the research and development of epitope peptide vaccine which can target all members of SSX family at the same time will have a wide application prospect. In this study, nine members of the SSX family were imported into the CTL epitope prediction software developed by Professor Lin Zhihua based on hypermotif and quantization sequence. The SYFPEITHI software named http: / www-bimas.cit.gov. / molbior-hla bindr.) and Heidelberg Biomedical Information Center in Germany rated the amino acid types of anchored residues and auxiliary residues on the basis of the amino acid types of specific peptide anchoring residues and auxiliary residues, and then analyzed their prediction results by using SYFPEITHI software named http: / www-bimas.cit.gov. / molbior.hla-bindr /) and the Heidelberg Biomedical Information Center in Germany, according to the amino acid types of the anchored and auxiliary residues of certain peptides. It was found that SSX57-65 was a peptide fragment with relatively high predictive score among all members, and the sequence similarity between them was very high, and only the difference was found in the first 89th position, while the sequence of other loci was completely consistent. On the basis of these results, we have synthesized four peptides representing all members of SSX57-65, I. e., 1) P1: AMTKLGFKA: SSX-2, SSX-3, SSX-5, SSX-7, SSX-9, 2) P4: AMTKLGFNV SSSX-6, SSX-8, P5) P5: AMTKLGFKV SSX-1, 4) P6: VMTKLGFKV. Based on the analysis of the binding affinity and binding stability of these four epitope peptides to MHC-I molecules, we used them to stimulate PBMCs in vitro and to evaluate the specific CTL.
【學位授予單位】：第三軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R392

【引證文獻】

相關(guān)期刊論文 前1條

1 韋力嘉;何少健;許淑茹;李希圣;;腫瘤—睪丸基因SSX1的原核表達載體的構(gòu)建[J];黑龍江醫(yī)藥;2011年01期

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本文編號：1986461