A bispecific diabody directed against prostate-specific membrane antigen and CD3 induces T-cell mediated lysis of prostate cancer cells.
A bispecific diabody directed against prostate-specific membrane antigen and CD3 induces T-cell mediated lysis of prostate cancer cells.
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DOI：
10.1007/s00262-007-0348-6
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发表时间：
2008-01
影响因子：
5.8
通讯作者：
Elsaesser-Beile, U.
中科院分区：
文献类型：
--
作者：
Buehler, P.;Wolf, P.;Gierschner, D.;Schaber, I.;Katzenwadel, A.;Schultze-Seemann, W.;Wetterauer, U.;Tacke, M.;Swamy, M.;Schamel, W. W. A.;Elsaesser-Beile, U.
尽管前列腺癌是男性最常诊断出的癌症之一，但在其进展超出可切除范围后，目前尚无治疗方法。因此，需要新的靶向治疗策略药物。通过双特异性单克隆抗体将肿瘤抗原与 T 细胞相关抗原交联已被证明可以增加 T 细胞中抗原特异性的细胞毒性。由于前列腺特异性膜抗原（PSMA）代表了一个极好的肿瘤靶标，因此双特异性双抗体的免疫疗法可能是前列腺癌的一种有前途的新型治疗选择。从抗 CD3 和抗 PSMA 单链 Fv 片段 (scFv) 的 DNA 构建对人 PSMA 和 T 细胞抗原 CD3 具有特异性的异二聚体双抗体。它使用含有双顺反子操纵子的载体在大肠杆菌中表达，以共分泌杂交 scFv VHCD3-VLPSMA 和 VHPSMA-VLCD3。通过固定化金属亲和层析 (IMAC) 从周质提取物中纯化所得的 PSMAxCD3 双抗体。通过流式细胞术在表达 PSMA 的前列腺癌细胞和 PSMA 阴性细胞系以及 Jurkat 细胞上测试了结合特性。对于体外功能分析，使用细胞活力测试（WST）。为了进行体内评估，将双抗体与人外周血淋巴细胞（PBL）一起应用于C4-2异种移植-SCID小鼠模型中。通过Blue Native凝胶电泳可以显示PSMAxCD3双抗体主要是四聚体。流式细胞术显示与表达 CD3 的 Jurkat 细胞和表达 PSMA 的 C4-2 细胞的特异性结合。在体外，双抗体被证明是重新靶向 PBL 以裂解 C4-2 前列腺癌细胞的有效试剂。用双抗体和 PBL 治疗接种 C4-2 肿瘤异种移植物的 SCID 小鼠可有效抑制肿瘤生长。 PSMAxCD3 双抗体具有促进前列腺癌免疫治疗和消除微小残留病的潜力。
Although cancer of the prostate is one of the most commonly diagnosed cancers in men, no curative treatment currently exists after its progression beyond resectable boundaries. Therefore, new agents for targeted treatment strategies are needed. Cross-linking of tumor antigens with T-cell associated antigens by bispecific monoclonal antibodies have been shown to increase antigen-specific cytotoxicity in T-cells. Since the prostate-specific membrane antigen (PSMA) represents an excellent tumor target, immunotherapy with bispecific diabodies could be a promising novel treatment option for prostate cancer. A heterodimeric diabody specific for human PSMA and the T-cell antigen CD3 was constructed from the DNA of anti-CD3 and anti-PSMA single chain Fv fragments (scFv). It was expressed in E. coli using a vector containing a bicistronic operon for co-secretion of the hybrid scFv VHCD3-VLPSMA and VHPSMA-VLCD3. The resulting PSMAxCD3 diabody was purified from the periplasmic extract by immobilized metal affinity chromatography (IMAC). The binding properties were tested on PSMA-expressing prostate cancer cells and PSMA-negative cell lines as well as on Jurkat cells by flow cytometry. For in vitro functional analysis, a cell viability test (WST) was used. For in vivo evaluation the diabody was applied together with human peripheral blood lymphocytes (PBL) in a C4-2 xenograft-SCID mouse model. By Blue Native gel electrophoresis, it could be shown that the PSMAxCD3 diabody is mainly a tetramer. Specific binding both to CD3-expressing Jurkat cells and PSMA-expressing C4-2 cells was shown by flow cytometry. In vitro, the diabody proved to be a potent agent for retargeting PBL to lyze C4-2 prostate cancer cells. Treatment of SCID mice inoculated with C4-2 tumor xenografts with the diabody and PBL efficiently inhibited tumor growth. The PSMAxCD3 diabody bears the potential for facilitating immunotherapy of prostate cancer and for the elimination of minimal residual disease.
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影响因子：
5.7
作者：
PERISIC, O;WEBB, PA;WILLIAMS, RL
通讯作者：
WILLIAMS, RL
影响因子：
6.4
作者：
Wang, SY;Diamond, DL;Vessella, RL
通讯作者：
Vessella, RL
影响因子：
254.7
作者：
Jemal, A;Tiwari, RC;Thun, MJ
通讯作者：
Thun, MJ
影响因子：
2.9
作者：
SCHAGGER, H;CRAMER, WA;VONJAGOW, G
通讯作者：
VONJAGOW, G
影响因子：
4.4
作者：
Cochlovius, B;Kipriyanov, SM;Little, M
通讯作者：
Little, M