(CRCL) vaccine for Chronic Myelogenous Leukemia

描述（由申请人提供）：我们的实验室开发了一种全新的方法来生产抗癌疫苗。利用自由溶液等电聚焦（FS-IEF）技术，我们可以有效地从肿瘤裂解物中富集多种伴侣复合物。已有文献证明，用这些富含伴侣蛋白的细胞裂解物（CRCL）接种疫苗比用纯化的单个伴侣蛋白（热休克蛋白，HSPs）接种疫苗更有效。通过将CRCL装载到树突状细胞（dc）上，可以进一步增强CRCL的抗原性，从而即使在已有疾病的情况下也能对小鼠肿瘤产生保护作用。除了它们的抗原携带能力外，CRCL对dc有强大的免疫刺激作用。作为佐剂，CRCL提供危险信号，增强药物治疗后发生凋亡的白血病细胞的免疫原性。慢性髓性白血病（CML）是研究慢性髓性白血病伴随肽抗原特异性免疫反应的一个有用模型。它在白血病中是独特的，因为bcr-abl癌基因本身是一种肿瘤抗原。此外，CML细胞表达其他潜在的靶抗原，如蛋白酶-3和Wilms肿瘤蛋白（WT1）。该提案的目标是产生足够强大的临床前数据，将CRCL疫苗推向临床环境。我们实验室打算继续在12B1小鼠CML模型中进行研究，以进一步了解CRCL疫苗的作用机制。与此同时，体外研究将检验人类cml衍生的CRCL对人类细胞的影响，以确定其有效性和安全性。提出以下具体目标。1)鉴定12B1小鼠白血病CRCL疫苗的肽抗原库。2)研究CRCL疫苗/佐剂与诱导小鼠12B1 bcr-abl+细胞凋亡的酪氨酸激酶抑制剂STI-571联合在体内的协同效应。3)人cml衍生CRCL的生化特征。4)评估人类cml衍生的CRCL对人类dc的影响，并检测CRCL脉冲dc产生白血病特异性ctl的潜力。这些目标的成功完成可能导致使用CRCL作为CML的有效疫苗，免疫疗法已经显示出对CML的希望。

DESCRIPTION (provided by applicant): Our laboratory has developed a completely new approach to generating an anticancer vaccine. Using a free solution isoelectric focusing (FS-IEF) technique we can efficiently enrich for multiple chaperone complexes from tumor lysates. It has been documented that vaccination with these chaperone rich cell lysates (CRCL) is more effective than immunization with purified individual chaperones (heat shock proteins, HSPs). The antigenicity of CRCL can be augmented further by loading them onto dendritic cells (DCs) resulting in protection against murine tumors even in the setting of pre-existing disease. In addition to their antigen carrying capacities, CRCL have potent immunostimulatory effects on DCs. As adjuvants CRCL provide danger signals enhancing the immunogenicity of leukemia cells undergoing apoptosis following drug treatment. Chronic Myelogenous Leukemia (CML) is a useful model for the study of antigen specific immune responses to peptides chaperoned by CRCL. It is unique among leukemias in that the bcr-abl oncogene itself is a tumor antigen. Moreover, CML cells express other potential target antigens such as Proteinase-3 and Wilms' tumor protein (WT1). The goal of this proposal is to generate sufficiently strong pre-clinical data to move CRCL vaccines into the clinical setting. Our laboratory intends to continue studies in the 12B1 murine CML model in order to understand further the mechanisms of action of CRCL vaccines. In parallel, in vitro studies will examine the effects of human CML-derived CRCL on human cells in order to establish efficacy and safety. The following specific aims are proposed. 1) Characterize the peptide antigen repertoire of CRCL vaccine derived from 12B1 murine leukemia. 2) Study the in vivo synergistic effects of combining CRCL vaccine/adjuvant with STI-571, a tyrosine kinase inhibitor that induces apoptosis in murine 12B1 bcr-abl+ cells. 3) Biochemically characterize human CML-derived CRCL. 4) Evaluate the effects of human CML-derived CRCL on human DCs and examine the potential of CRCL-pulsed DCs to generate leukemia specific CTLs. The successful completion of these aims may result in the use of CRCL as an effective vaccine against CML, a disease in which immunotherapy has already shown promise.