ANTI-IDIOTYPE CYTOKINE FUSION PROTEIN AS TUMOR VACCINE

作为我们长期目标的一部分， 我们在此提出的肿瘤疫苗， 用于治疗乳腺癌患者。 我们已选定为“第一 第二代”Ab 2，11 D10“，其用作特定细胞的内部图像。 人乳脂球（HMFG）的表位，并且是治疗性Ab 2 在项目I中使用。 “第二代”Ab 2将由 与关键细胞因子的联系。 细胞因子已被证明可以增加 包括独特型在内的蛋白质抗原的免疫原性。 我们将使用两个 细胞因子IL-2和GM-CSF，以产生11 D10的嵌合抗体， 将含有连接到恒定区的两种细胞因子中的任一种 （CH3）。 将克隆1211 D10的H链基因的可变外显子， 测序 VH基因将被整合到表达 该恒定基因与IL-2或GM-CSF的基因融合。 的重 11 D10杂交瘤细胞系的链丢失突变体将用 这些质粒。 功能性细胞因子和IG的表达将被 在转染瘤克隆中监测， 选定. 这些嵌合抗体将在小鼠和非人类中进行测试。 灵长类动物作为抗独特型肿瘤疫苗：1）动物中的Ab 3滴度 将用嵌合抗体免疫的小鼠的Ab 3滴度与用嵌合抗体免疫的小鼠的Ab 3滴度比较。 用项目1中提出的明矾沉淀的Ab 2免疫的动物。 2)还将在这些动物组中比较Ab 1 '。 我们预计 为了观察嵌合Ab 2中显著更高的Ab 3和Ab 1 '滴度， 免疫组。 我们还将比较在小鼠中对Ab 2的增殖反应。 两组。 如果这些嵌合细胞因子Ab 2的临床前研究 在动物中诱导上级抗TAA应答，我们将启动I期 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.