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细胞毒性T淋巴细胞(又称杀伤性T淋巴细胞,cytotoxic T lymphocytes, CTL),是一种特异T细胞,分泌各种细胞因子参与免疫作用。对某些病毒、肿瘤细胞具有杀伤作用,与自然杀伤细胞构成机体抗病毒、抗肿瘤免疫的重要防线。CTL是肿瘤免疫过继疗法主要效应细胞之一。我们的CTL细胞治疗技术是利用专有技术制备的人工抗原提呈细胞,能高效负载多靶点的多种肿瘤抗原肽,起到高效刺激T细胞的作用,比目前国内外用的自体DC细胞或其他各种人工抗原提呈细胞株有更强的刺激CD8T细胞的效果,技术优势明显且安全性高。公司的专有技术是从美国强生获取,并在大中华地区使用,我们创新地应用这个技术开展了对新的肿瘤适应症的研究。

肿瘤抗原肽是新一类肿瘤标志物。肿瘤抗原在细胞体内被分解后产生的小分子多肽,和MHC I结合形成MHC I-抗原肽复合物后,表达在癌细胞的表面。该复合物能够被CD8 T细胞识别,因此可以被利用作为癌细胞的靶分子,可用于诊断和治疗的各个方面,因此鉴定癌细胞肿瘤抗原肽具有重要的意义。我们发展出了一套综合了免疫-生物化学化/生物物理-生物信息学方法来鉴定肿瘤抗原肽的技术平台。细胞学试验表明,鉴定出来的抗原肽能诱导出高度特异的肿瘤杀伤作用。技术特点l  自主知识产权,已授权。(**号:201310439062.2);l  该方法得到的抗原肽是直接经细胞处理加工的抗原肽,真实有效;l  抗原肽确定后可刺激CD8T细胞产生有肿瘤杀伤功能的CTL,可用于细胞治疗;l  抗原肽所对应抗原可作为肿瘤标志物,用于肿瘤诊断试剂的研发(抗体诊断、PCR诊断);l  肿瘤新抗原疫苗的研发。Tumor antigen peptide identification platform    Tumor antigen peptide is a new class of tumor markers. Tumor antigens are decomposed in the cell to generate peptides, which bind to MHC-1 to form MHC-1/antigen peptide complexes, and are displayed on cancer cell surface. The complexes can be recognized by corresponding CD8 T cells, and therefore can be used as targets for diagnosis and treatment. Identification of cancer cell tumor antigen peptides is, therefore, of great importance. We have developed a technical platform that integrates immune-biochemical / biophysical-bioinformatics methods to identify tumor antigen peptides. Our studies on cultured cells have shown that the identified antigenic peptides can induce a highly specific tumor killing effect.  Technical features and highlights1.    Patented technology involved (patent no. 201310439062.2);2.    The peptides thus obtained are those naturally processed by tumor celss, therefore        authantic and useful;3.    The peptide(s) can be used to stimulate tumor specifc CD8 T cell for cell immunotherapy;4.    The peptide(s) origing protein(s) can be the tumor marker(s),  of potential use in diagnoisis;5.    Useful in anti-tumor peptide vaccine development.

将已构建的插入IL-24等细胞因子编码基因的溶瘤病毒,转染到体外诱导的CIK细胞内,通过特殊的基因改造,使溶瘤病毒在CIK细胞内增殖但不破坏CIK细胞,然后向患者体内回输CIK细胞,使融瘤病毒特异性导向肿瘤细胞,在其内大量复制并*终摧毁肿瘤细胞。技术特点l   CIK细胞本身具有非MHC限制性的非特异性免疫杀伤作用,对肿瘤细胞有杀伤活性;l   通过CIK细胞携带溶瘤病毒并释放到肿瘤细胞内,使溶瘤病毒特异性导向肿瘤细胞,并发挥溶瘤活性;l   插入在溶瘤病毒基因组的细胞因子基因随着病毒的扩增在肿瘤细胞内表达,并释放细胞因子,直接发挥抗肿瘤活性;l   CIK细胞作为载体向肿瘤细胞导向溶瘤病毒,减少溶瘤病毒直接注射被机体免疫系统的清除以及副作用;    因此这种溶瘤病毒与CIK免疫细胞的联合治疗法比单一的细胞治疗或溶瘤病毒治疗具有更高的特异性和杀伤活性。Oncolytic virus / CIK technologyThe oncolytic virus integrated with IL-24 and other cytokine genes was transduced into CIK cells. The genetically modified oncolytic virus would proliferate in CIK cells without destroying CIK cells. The virus harbored CIK cells were then transfused to the patient, specifically targeting the oncolytic virus to tumor cells, and ultimately destroy the tumor cells. Technical features 1.   CIK cells possess non-MHC-specific and nonspecific cytotoxicity against tumor cells; 2.   Oncolytic virus carried in the CIK cells would be targeted to, released into and then more       efficiently kill the tumor cells;3.   The cytokine gene integrated in the genome of the oncolytic virus is expressed in the tumor       cells along with the expansion of the virus, which directly develop the antitumor activity. 4.   Using CIK cell as carrier of oncolytic virus helps to reduce the clearence of virus by receptor       immune system and otherwise some side effects caused by naked virus injectionTherefore, the combination of the oncolytic virus and CIK immune cells has a higher specificity and killing activity than CIK or oncolytic virus alone.

双特异性抗体是含有两种特异性抗原结合位点的双功能抗体。我们通过化学交联法将抗CD3单抗与曲妥珠单抗交联,获得了对肿瘤细胞和淋巴细胞的双向识别作用的双特异抗体。此双特异性抗体一方面通过CD3单抗识别CIK细胞;另一方面,通过曲妥珠单抗识别肿瘤表面HER2受体。在DC-CIK细胞体外诱导、培养的**阶段加入该双特异抗体,双特异抗体即通过抗CD3抗体结合到CIK细胞表面,CIK细胞回输到体内后,其表面的双特异抗体将CIK细胞靶向募集到肿瘤细胞表面,从而提高CIK细胞对肿瘤细胞的识别和杀伤活性。技术特点l   制备技术相对简单;l   利用抗体-抗原的强大亲和力,大大提高DC-CIK细胞对肿瘤细胞的识别和杀伤机会。Bispecific antibody / DC-CIK technologyA bispecific antibody is a bifunctional antibody that contains two specific antigen binding sites. We cross-linked anti-CD3 monoclonal antibody with trastuzumab by chemical cross-linking, and obtained bi-specific antibodies for bi-directional recognition of tumor cells and lymphocytes. This bispecific antibody recognizes CIK cells via CD3 target and recognizes tumor cells via HER2 receptor. In the final stage of DC-CIK cells culturing, the bispecific antibody was added and binds to the surface of CIK cells through anti-CD3 antibody. After transfusion, the  cells were recruited to tumor cells through bispecific antibody, thereby enhancing the recognition and killing activity of CIK cells to tumor cells. Technical features1.   The preparation technique is relatively simple 2.   The strong affinity of antibody to antigen greatly improves the DC-CIK cells on tumor       cell recognition and killing opportunities

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