(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产生抗癌疫苗。使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分子伴侣复合物。已经证明，用这些富含分子伴侣的细胞裂解物（CRCL）接种比用纯化的单个分子伴侣（热休克蛋白，HSP）免疫更有效。CRCL的抗原性可以通过将其加载到树突状细胞（DC）上进一步增强，从而即使在预先存在的疾病的情况下也能保护小鼠免受肿瘤的侵害。CRCL除了具有抗原携带能力外，还对DC具有强的免疫刺激作用。作为佐剂，CRCL提供危险信号，增强药物治疗后经历凋亡的白血病细胞的免疫原性。慢性粒细胞性白血病（CML）是研究CRCL分子伴侣肽抗原特异性免疫应答的有效模型。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（WT 1）。该提案的目标是生成足够强的临床前数据，以将CRCL疫苗推向临床环境。我们的实验室打算继续在12 B1小鼠CML模型中进行研究，以进一步了解CRCL疫苗的作用机制。同时，体外研究将检查人CML衍生的CRCL对人细胞的影响，以确定疗效和安全性。提出了以下具体目标。1)表征源自12 B1小鼠白血病的CRCL疫苗的肽抗原库。2)研究CRCL疫苗/佐剂与STI-571（一种诱导小鼠12 B1 bcr-abl+细胞凋亡的酪氨酸激酶抑制剂）组合的体内协同效应。3)对人CML衍生CRCL进行生物化学表征。4)评估人CML衍生的CRCL对人DC的作用，并检查CRCL脉冲的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CRCL作为针对CML的有效疫苗的使用，CML是一种免疫疗法已经显示出希望的疾病。

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.