Immuno-gene therapy by secreted gp96-Ig fusion protein

分泌型 gp96-Ig 融合蛋白的免疫基因治疗

• 批准号: 13670584
• 负责人: YAMAZAKI Koichi
• 金额: $ 2.3万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670584/
• 关键词: secreted heat shock fusion protein; gp96; immuno-gene therapy; CD8 CTL; tumor rejection

Gp96 has useful properties as tumor vaccine for the generation of CD8 CTL independent of MHC restriction. We have developed a secreted form of gp96-Ig (gp96-Ig) by deleting the endoplasmic reticulum retention signal, KDEL, and replacing it with the Fc portion of murine IgG_1. Tumor cells transfected with and overexpressing gp96-Ig were less tumorigenic, compared to wild type tumors. Immunization of mice with the murine lymphoma E.G7 transfected with gp96-Ig (E.G7-gp96-Ig), but not with irradiated E.G7, induced immunity to subsequent challenge with E.G7. Depletion studies showed that CD8^+ cells were required for E.G7-gp96-Ig rejection throughout induction phase and effector phase, but not CD4^+ cells or macrophages. In this study, we examined the immunotherapeutical potency by gp96-Ig gene-transfected tumor cells. E.G7 was established in C57BL/6 mice for three days. Daily injections, beginning on day 4, of gp96-Ig-transduced tumor cells suppressed tumor growth remarkably when compared … More to PBS treatment or to treatment with control tumors treatment with control tumors secreting gp96-Ig. Beginning vaccination on day 5 after tumor transplantation, successful therapy required a vaccination schedule of two daily injections of E.G7-gp96-lg. In a Jung tumor model. LLC, LLC-gp96-Ig vaccines were equally effective as gp96 purified from LLC cells and comparable to IL-12-transduced tumor cells (LLC/1L12). There were no synergistic effects by a 1:1 mixture of LLC-gp96-Ig and LLC/IL12, suggesting that the two vaccines may act through similar cellular mechanisms. CD4^+ T cells were not required for vaccines by gp96-Ig-transduced tumor cells. CD8^+ cytotoxic activities specific for wild type tumor cells were induced by vaccines with gp96-Ig-transduced tumor cells. Moreover, adoptively transferred, ovalbumin specific T-cell receptor (TCR) transgenic CD8^+ cells (OT-1) responded with clonal expansion to the immunization with EG7-gp96-Ig. Our data suggest that tumors secreting gp96-Ig may be useful as potent anti tumor vaccines. Less

Gp 96具有作为肿瘤疫苗的有用特性，用于产生不依赖于MHC限制的CD 8 CTL。我们通过删除内质网滞留信号KDEL并用鼠IgG_1的Fc部分取代它，开发了分泌形式的gp 96-IG（gp 96-IG）。与野生型肿瘤相比，用gp 96-IG转染并过表达的肿瘤细胞的致瘤性较低。用gp 96-IG转染的鼠淋巴瘤E.G7（E.G7-gp 96-IG）而不是用辐照的E.G7免疫小鼠，诱导对随后用E.G7攻击的免疫。耗竭研究表明，在整个诱导期和效应期，E.G7-gp 96-IG排斥反应需要CD 8 ^+细胞，而不是CD 4 ^+细胞或巨噬细胞。在本研究中，我们通过gp 96-IG基因转染的肿瘤细胞来检测免疫治疗效力。在C57 BL/6小鼠中建立E.G7三天。从第4天开始，每天注射gp 96-Ig转导的肿瘤细胞显著抑制肿瘤生长， ...更多信息 PBS处理或用分泌gp 96-IG的对照肿瘤处理。在肿瘤移植后第5天开始疫苗接种，成功的治疗需要在Jung肿瘤模型中每天注射两次E.G7-gp 96-Ig的疫苗接种方案。LLC，LLC-gp 96-IG疫苗与从LLC细胞纯化的gp 96同样有效，并且与IL-12转导的肿瘤细胞（LLC/1 L12）相当。LLC-gp 96-IG和LLC/IL 12的1：1混合物没有协同效应，表明这两种疫苗可能通过相似的细胞机制起作用。gp 96-Ig转导的肿瘤细胞的疫苗不需要CD 4 ^+ T细胞。用gp 96-Ig转导的肿瘤细胞制成的疫苗可诱导特异性针对野生型肿瘤细胞的CD 8 ^+细胞毒活性。此外，过继转移的卵清蛋白特异性T细胞受体（TCR）转基因CD 8 ^+细胞（OT-1）对EG 7-gp 96-IG免疫的反应是克隆扩增。我们的数据表明，肿瘤分泌gp 96-IG可能是有用的有效的抗肿瘤疫苗。少

项目成果

Hommura F, Furuuchi K, Yamazaki K, Ogura S, Kinoshita I, Shimizu M, Moriuchi T, Katoh H, Nishimura M, Dosaka-Akita H: "Increased expression of β-catenin predicts better prognosis in nonsmall cell lung carcinoma"Cancer. 94. 752-758 (2002)

Hommura F、Furuuchi K、Yamazaki K、Ogura S、Kinoshita I、Shimizu M、Moriuchi T、Katoh H、Nishimura M、Dosaka-Akita H：“β-连环蛋白表达增加可预测非小细胞肺癌更好的预后”癌症。 94.752-758 (2002)

山崎浩一: "Annual Review免疫2001"中外医学社編. 308-315 (2002)

Koichi Yamazaki：“免疫学年度评论 2001”，由 Chugai Igakusha 编辑 308-315 (2002)。

Dosaka-Akita H, Kinoshita I, Yamazaki K, Izumi H, Itoh T, Katoh H, Nishimura M, Matsuo K, Yamada Y, Kohno K: "N-acetylgalactosaminy1 transferase-3 (GalNAc-T3) is a potential new marker for non-small cell lung cancers"Br J Cancer. 87. 751-755 (2002)

Dosaka-Akita H、Kinoshita I、Yamazaki K、Izumi H、Itoh T、Katoh H、Nishimura M、Matsuo K、Yamada Y、Kohno K：“N-乙酰半乳糖胺 1 转移酶 3 (GalNAc-T3) 是一种潜在的新标记物

Dosaka-Akita H, Kinoshita I, Yamazaki K, Izumi H, Itoh T, Katoh H, Nishimura M, Matsuo K, Yamada Y, Kohno K: "N-acetylgalactosaminyl transferase-3 (GalNAc-T3) is a potential new marker for non-small cell lung cancers"Br J Cancer. 87. 751-755 (2002)

Dosaka-Akita H、Kinoshita I、Yamazaki K、Izumi H、Itoh T、Katoh H、Nishimura M、Matsuo K、Yamada Y、Kohno K：“N-乙酰半乳糖胺基转移酶 3 (GalNAc-T3) 是一种潜在的新标记物

Masutani M, Suzuki J, Matsuda T, Dochin A, Sadaoka K, Nomura A, Ohira K, Takahashi T, Yamazaki K, Dosaka-Akita H, Nishimura M, Kawakami Y.: "Increased apoptosis associated with depressed type of early intestinal gastric cancer"Jpn J Cancer Res. 92. 1214-1

Masutani M、Suzuki J、Matsuda T、Dochin A、Sadaoka K、Nomura A、Ohira K、Takahashi T、Yamazaki K、Dosaka-Akita H、Nishimura M、Kawakami Y.：“与早期肠胃抑郁型相关的细胞凋亡增加