CONSTRUCTION OF VACCINES AGAINST BREAST CANCER

这项提议的总体目标是确定抗原表位 对人体免疫系统具有免疫原性的乳腺癌，决定了 可作为第三阶段基础的最佳受益方法 疫苗接种试验。两个乳腺癌细胞系，选择高表达 Lewis(Y)、Lewis(X)、Tf、STN、MUC-1、CEA、HER2/neu、ras和 突变型P53，将作为免疫原。它们的免疫原性将是 免疫佐剂QS21和IL-2基因转导的增强作用 和干扰素伽马。将检测这些抗原的免疫原性 最初通过标准的抗体测试，迟发性超敏反应 和针对这些抗原的增殖性T细胞反应，以及人类白细胞抗原 针对MUC-1的非限制性细胞毒T细胞反应。的敏感性 人类单抗的建立将进一步提高检测水平 抗体及体外致敏和抗原特异性克隆 T细胞系。大多数这些抗原的纯化和/或合成版本 此外，还有一组分型细胞系可用于这些检测。 一旦抗原被确定为潜在的免疫原性(和 确定免疫原性表位)锁孔帽状血蓝蛋白(KLH)结合物 疫苗将在诊所准备和测试。既然有三个乳房 已知癌症抗原对人类具有潜在的免疫原性。 (Tf、STN和MUC-1)这些将用于初始的KLH结合物 疫苗试验。我们的具体目标是： 1.比较乳腺癌表达的肿瘤抗原的免疫原性 照射后的细胞株SKBR3和MCF7 免疫佐剂QS21，已转导IL-2和 干扰素-伽马基因，或与这些细胞因子一起转导的基因 基因，并与QS21混合。 2.确定最佳合成表位和结合物制剂 免疫原性乳房的抗体诱导和T细胞反应 当癌症抗原被识别时。最初，这将包括Tf， STN和MUC-1抗原。 3.比较人乳腺癌抗原表达的免疫原性。 在AIM I中筛选出具有免疫原性的最佳全细胞疫苗 这些相同的抗原在最佳合成结合疫苗中表达 在目标2中确定。 4.使用最具免疫原性的疫苗组合进行第二阶段试验 在30名乳腺癌患者中，他们没有发现疾病，但 循环中的肿瘤标志物升高。

The overall goals of this proposal are to identify antigenic epitopes on breast cancer which are immunogenic to the human immune system, determine the optimal method of benefit which may be the basis for a Phase III vaccination trial. Two breast cancer cell lines, selected for high expression of Lewis(y), Lewis(X), Tf, sTn, MUC-1, CEA, HER2/neu, ras and mutant p53, will be used as immunogen. Their immunogenicity will be boosted by immunological adjuvant QS21 and transduction of genes for IL2 and interferon gamma. Immunogenicity of these antigens will be detected initially by standard tests for antibody, delayed type hypersensitivity and proliferative T cell responses against each of these antigens, and HLA unrestricted cytotoxic T cell responses against MUC-1. The sensitivity of detection will be further increased by establishment of human monoclonal antibodies and by in vitro sensitization and cloning of antigen specific T cell lines. Purified and/or synthetic versions of most of these antigens as well as a panel of typing cell lines are available for these assays. Once antigens are identified as potentially immunogenic (and the immunogenic epitopes determined) keyhole limpet hemocyanin (KLH) conjugate vaccines will be prepared and tested in the clinic. Since three breast cancer antigens are already known to be potentially immunogenic in man (Tf, sTn and MUC-1) these will be used in the initial KLH conjugate vaccine trials. Our specific aims are: 1. Compare the immunogenicity of tumor antigens expressed on breast cancer cell lines SKBR3 and MCF7 which have been irradiated and mixed with immunological adjuvant QS21, which have been transduced with the IL2 and interferon gamma genes, or which have been transduced with these cytokine genes and mixed with QS21. 2. Determine the optimal synthetic epitopes and conjugate preparations for induction of antibody and T cell responses against immunogenic breast cancer antigens as they are identified. Initially this will include Tf, sTn and MUC-1 antigens. 3. Compare the immunogenicity of breast cancer antigens expressed on the optimal whole cell vaccine identified in Aim I with the immunogenicity of these same antigens expressed in the optimal synthetic conjugate vaccines identified in Aim 2. 4. Conduct a Phase II trial with the most immunogenic vaccine combination in 30 patients with breast cancer who are free of detectable disease but have elevated circulating tumor markers.