Establishment of tumor immunotherapy using class II

II类肿瘤免疫疗法的建立

• 批准号: 14570154
• 负责人: YAZAWA Takuya
• 金额: $ 2.24万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570154/
• 关键词: Class II Transactivator; Major Histocompatibility Complex; Cancer; Tumor Immunity; Tatタンパク

In the first year, we tried to establish the system to delivery the recombinant hCIITA molecules into cancer cells. We first used VP22 tag derived from HSV as a candidate of deliverers, the VP22 interfered the function of hCIITA. Thus, we next used Tat tag derived from HIV and obtained the facts that Tat-hCIITA has enough function to induce MHC antigens in cancer cells and purified recombinant Tat-hCIITA protein in the culture media move into cancer cells and could make MHC induce. However, it was impossible to get enough amounts of recombinant Tat-hCIITA protein, since the protein formed inclusion body in E.coli.In the second year, we tried to establish the co-expression system of hCIITA and MHC haplotype-matched antigenicoligopeptides (Tb (k) and Tb (km) = I-A^k-haplotype matched oligopeptides, Tb (b) = I-A^b-haplotype matched oligopeptides) derived from Mycobacterium tuberculosis alpha antigen. We constructed the expression vectors, pZeo-hCIITA-HA-furin-Tb (k) -His, pZeo-hCIITA-HA furin-Tb (km) -His, and pZeo-hCIITA-HA-furin-Tb (b)-His and transfected into he C1300 (mouse neuroblastoma cell line derived from A/J mouse (I-A^k haplotype), MM102 (mouse breast cancer cell line derived from C3H/HeN mouse (I-A^k haptotype), and Lewis (mouse lung cancer cell line derived from C57BL/6 mouse (I-A^b haplotype). We have established stably hCIITA-and Tb (b)-coexpressed transfectants (Lewis-pZeo-hCIITA-HA-furin-Tb(b)-His and MM102-pZeo-hCIITA-HA furin-Tb (b)-His) and got the evidence that Lewis-pZeo-hCIITA-HA furin-Tb (b)-His generates well tumor immunity against tumor-laden hosts.

第一年，我们尝试建立将重组hCIITA分子递送到癌细胞中的系统。我们首先使用源自HSV的VP22标签作为候选传递者，VP22干扰hCIITA的功能。因此，我们接下来使用源自HIV的Tat标签，并获得事实：Tat-hCIITA具有足够的功能来诱导癌细胞中的MHC抗原，并且培养基中纯化的重组Tat-hCIITA蛋白进入癌细胞并可以诱导MHC。然而，由于重组Tat-hCIITA蛋白在大肠杆菌中形成包涵体，不可能获得足够量的重组蛋白。第二年，我们尝试建立hCIITA和MHC单倍型匹配的抗原寡肽的共表达系统（Tb（k）和Tb（km）= I-A^k-单倍型匹配的寡肽，Tb（b）） = I-A^b-单倍型匹配的寡肽）源自结核分枝杆菌 α 抗原。我们构建了表达载体 pZeo-hCIITA-HA-furin-Tb (k) -His、pZeo-hCIITA-HAfurin-Tb (km) -His 和 pZeo-hCIITA-HA-furin-Tb (b)-His 并转染至 he C1300（源自 A/J 小鼠（I-A^k 单倍型）的小鼠神经母细胞瘤细胞系 MM102） (来自C3H/HeN小鼠的小鼠乳腺癌细胞系(I-A^k单倍型)和Lewis(来自C57BL/6小鼠的小鼠肺癌细胞系(I-A^b单倍型)。我们已经建立了稳定的hCIITA-和Tb (b)-共表达转染子(Lewis-pZeo-hCIITA-HA-furin-Tb(b)-His和 MM102-pZeo-hCIITA-HA Furin-Tb (b)-His) 并得到证据表明 Lewis-pZeo-hCIITA-HA Furin-Tb (b)-His 对荷瘤宿主产生良好的肿瘤免疫。

项目成果

Yazawa T: "Class II transactivator (CIITA) deficiency in tumor cells : Complicated mechanisms or not?"Am J Pathol. 163. 373-376 (2003)

Yazawa T：“肿瘤细胞中 II 类反式激活因子 (CIITA) 缺陷：机制是否复杂？”Am J Pathol。

Okudela K: "K-ras gene mutation enhances motility of immortalized airway cells and lung adenocarcinoma cells via AKT activation"Am J Pathol. 164. 91-100 (2004)

Okudela K：“K-ras 基因突变通过 AKT 激活增强永生化气道细胞和肺腺癌细胞的运动性”Am J Pathol。

Yazawa T: "Complicated mechanisms of class II transactivator transcription deficiency in small cell lung cancer and neuroblastoma"Am J Pathol. 161. 291-300 (2002)

Yazawa T：“小细胞肺癌和神经母细胞瘤中 II 类反式激活因子转录缺陷的复杂机制”Am J Pathol。

Yazawa T, et al.: "Colliding primary lung cancers of adenosquamous carcinoma and large cell neuroendocrine carcinoma."Pathol Int. 53. 58-65 (2003)

Yazawa T 等人：“腺鳞癌和大细胞神经内分泌癌的原发性肺癌的冲突。”Pathol Int。

Yazawa T, et al.: "Complicated mechanisms of class II transactivator transcription deficiency in small cell lung cancer and neuroblastoma."Am J Pathol. 161. 291-300 (2002)

Yazawa T 等人：“小细胞肺癌和神经母细胞瘤中 II 类反式激活因子转录缺陷的复杂机制。”Am J Pathol。