Sen M, Wankowski D M, Garlie N K, Siebenlist R E, Van Epps D, LeFever A V, Lum L G
Blood Center of Southeastern Wisconsin, Milwaukee, WI 53201, USA.
J Hematother Stem Cell Res. 2001 Apr;10(2):247-60. doi: 10.1089/15258160151134944.
Relapse after adjuvant chemotherapy or high-dose chemotherapy with stem cell transplant for high-risk breast cancer remains high and new strategies that provide additional antitumor effects are needed. This report describes methods to generate highly effective HER2/neu-specific cytotoxic T cells by arming activated T cells with anti-CD3 x anti-HER2/neu bispecific antibody (BsAb). OKT3 and 9184 (anti-HER2) monoclonal antibodies (mAb) were conjugated and used to arm T cells that were subsequently tested in binding, cytotoxicity, and cytokine secretion assays. Armed T cells aggregated and specifically killed HER2/neu(+) breast cancer cells. Cytotoxicity emerged after 6 days of culture, was higher in armed T cells than unarmed T cells at all effector to target ratios (E/T) tested, and increased as the arming dose was increased. At an E/T of 20:1, the mean cytotoxicity of armed activated T cells (ATC) from 10 normal subjects increased by 59 +/- 11% (+/-SD) over that seen in unarmed ATC (p < 0.001) and the mean cytotoxicity of armed ATC from 6 cancer patients increased by 32 +/- 9% above that seen for unarmed ATC (p < 0.0004). After arming, the BsAb persisted on ATC up to 72 h and armed ATC continued to be cytotoxic up to 54 h. The amount of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and granulocyte-macrophage colony-stimulating factor (GM-CSF) secreted was 1699, 922, and 3092 pg/ml/10(6) cells per 24 h, respectively, when armed T cells were exposed to a HER2/neu(+) breast carcinoma cell line. These studies show the feasibility and clinical adaptability of this approach for generating large numbers of anti-HER2-specific, cytotoxic T cells for clinical trials.
高危乳腺癌患者在接受辅助化疗或高剂量化疗联合干细胞移植后复发率仍然很高，因此需要能提供额外抗肿瘤作用的新策略。本报告描述了通过用抗CD3×抗HER2/neu双特异性抗体（BsAb）武装活化T细胞来生成高效HER2/neu特异性细胞毒性T细胞的方法。将OKT3和9184（抗HER2）单克隆抗体（mAb）偶联，用于武装T细胞，随后在结合、细胞毒性和细胞因子分泌试验中对其进行检测。武装后的T细胞聚集并特异性杀伤HER2/neu(+)乳腺癌细胞。培养6天后出现细胞毒性，在所有测试的效应细胞与靶细胞比例（E/T）下，武装T细胞的细胞毒性均高于未武装T细胞，且随着武装剂量的增加而增加。在E/T为20:1时，10名正常受试者的武装活化T细胞（ATC）的平均细胞毒性比未武装ATC增加了59±11%（±标准差）（p<0.001），6名癌症患者的武装ATC的平均细胞毒性比未武装ATC增加了32±9%（p<0.0004）。武装后，BsAb在ATC上持续存在长达72小时，武装后的ATC在长达54小时内仍具有细胞毒性。当武装T细胞暴露于HER2/neu(+)乳腺癌细胞系时，每24小时分泌的干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）的量分别为1699、922和3092 pg/ml/10(6)个细胞。这些研究表明了该方法用于生成大量抗HER2特异性细胞毒性T细胞用于临床试验的可行性和临床适应性。
