ANTI-IDIOTYPE CYTOKINE FUSION PROTEIN AS TUMOR VACCINE

作为我们长期目标的一部分，用抗独特型药物治疗癌症患者 我们在这里提出的产生“第二代Ab2”的肿瘤疫苗 用于乳腺癌患者的治疗。我们被选为“第一名” 第“代”AB2、11D10，用作特定 人乳脂肪小球(HMFG)的表位，是治疗性AB2 在项目I中使用。“第二代”AB2将由 与关键细胞因子的联系。细胞因子已被证明可以增加 蛋白质抗原的免疫原性，包括独特型。我们将使用两个 细胞因子IL-2和GM-CSF，以产生11D10的嵌合抗体，该抗体 将包含与恒定结构域连接的两种细胞因子中的任何一种 (CH3)。1211D10的H链基因的可变外显子将被克隆并 已排序。VH基因将被整合到表达 恒定基因与IL-2或GM-CSF基因融合。一个沉重的 11D10杂交瘤细胞株的链缺失突变株将被 这些质粒。功能性细胞因子和免疫球蛋白的表达 将在转染瘤克隆和高产克隆中进行监测 被选中了。这些嵌合抗体将在老鼠和非人类身上进行测试。 灵长类作为抗独特型肿瘤疫苗：1)动物体内的ab3滴度 用嵌合抗体免疫，将与ab3滴度进行比较 按照项目1的建议，用明矾沉淀的AB2免疫的动物。 2)AB1‘也将在这些动物群体中进行比较。我们预计 在嵌合的ab2中观察到显著更高的ab3和ab1‘滴度 免疫组。我们还将比较AB2的增殖反应在 两组都是。如果这些嵌合细胞因子Ab2s的临床前研究 在动物身上诱导出优越的抗TAA反应我们将启动一个阶段 乳腺癌患者的临床试验。

As part of our long-term goals to treat cancer patients with anti-idiotypic tumor vaccines we are proposing here to generate "Second Generation Ab2s" for the therapy of breast cancer patients. We have selected as the "First Generation" Ab2, 11D10, which functions as an internal image for a specific epitope of the human milk fat globule (HMFG) and is the therapeutic Ab2 used in Project I. The "Second Generation" Ab2 will be constructed by linkages to key cytokines. Cytokines have been shown to increase the immunogenicity of protein antigens including idiotypes. We will use two cytokines, IL-2 and GM-CSF, to create a chimeric antibody of 11D10, which will contain either of the two cytokines attached to the constant domain (CH3). The variable exon of the H chain gene of 1211D10 will be cloned and sequenced. The VH gene will be incorporated into plasmids which express the constant gene fused to the genes for either IL-2 or GM-CSF. A heavy chain loss mutant of the 11D10 hybridoma cell line will be transfected with these plasmids. Expression of functional cytokines and Ig will be monitored in transfectoma clones and high producing clones will be selected. These chimeric antibodies will be tested in mice and nonhuman primates as anti-idiotype tumor vaccines: 1) the Ab3 titer in animals immunized with the chimeric antibody will be compared to the Ab3 titer in animals immunized with the alum precipitated Ab2 as proposed in Project 1. 2) The Ab1' will also be compared in these groups of animals. We expect to observe a significantly higher Ab3 and Ab1' titer in the chimeric Ab2 immunized group. We also will compare the proliferative response to Ab2 in both groups. If these preclinical studies with chimeric cytokine Ab2s induce superior anti-TAA responses in animals we will initiate a phase I clinical trial in breast cancer patients.