McCormick A A, Kumagai M H, Hanley K, Turpen T H, Hakim I, Grill L K, Tusé D, Levy S, Levy R
Biosource Technologies, Inc., 3333 Vacavalley Parkway, Suite 1000, Vacaville, CA 95688, USA.
Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):703-8. doi: 10.1073/pnas.96.2.703.
Rapid production of protein-based tumor-specific vaccines for the treatment of malignancies is possible with the plant-based transient expression system described here. We created a modified tobamoviral vector that encodes the idiotype-specific single-chain Fv fragment (scFv) of the immunoglobulin from the 38C13 mouse B cell lymphoma. Infected Nicotiana benthamiana plants contain high levels of secreted scFv protein in the extracellular compartment. This material reacts with an anti-idiotype antibody by Western blotting, ELISA, and affinity chromatography, suggesting that the plant-produced 38C13 scFv protein is properly folded in solution. Mice vaccinated with the affinity-purified 38C13 scFv generate >10 micrograms/ml anti-idiotype immunoglobulins. These mice were protected from challenge by a lethal dose of the syngeneic 38C13 tumor, similar to mice immunized with the native 38C13 IgM-keyhole limpet hemocyanin conjugate vaccine. This rapid production system for generating tumor-specific protein vaccines may provide a viable strategy for the treatment of non-Hodgkin's lymphoma.
利用本文所述的基于植物的瞬时表达系统,能够快速生产用于治疗恶性肿瘤的基于蛋白质的肿瘤特异性疫苗。我们构建了一种修饰的烟草花叶病毒载体,其编码来自38C13小鼠B细胞淋巴瘤免疫球蛋白的独特型特异性单链Fv片段(scFv)。被感染的本氏烟草植株在细胞外区室中含有高水平的分泌型scFv蛋白。该物质通过蛋白质印迹法、酶联免疫吸附测定和亲和色谱法与抗独特型抗体发生反应,表明植物产生的38C13 scFv蛋白在溶液中正确折叠。用亲和纯化的38C13 scFv免疫的小鼠产生>10微克/毫升的抗独特型免疫球蛋白。这些小鼠受到同基因38C13肿瘤致死剂量攻击时受到保护,类似于用天然38C13 IgM-钥孔血蓝蛋白缀合疫苗免疫的小鼠。这种用于生成肿瘤特异性蛋白质疫苗的快速生产系统可能为非霍奇金淋巴瘤的治疗提供一种可行的策略。