Cryopreparation of PBMC for Immunotherapy and Immune Assessment Studies.

用于免疫治疗和免疫评估研究的 PBMC 冷冻制备。

• 批准号: 7617659
• 负责人: JOHN R. YANNELLI
• 金额: $ 18.2万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7617659
• 关键词: Acute; Address; Aftercare; Antibodies; Area; Biological Assay; Blood; Boxing; Cell Survival; Cell physiology; Cells; Cellular Immunity; Clinical; Clinical Research; Clinical Trials; Cryopreservation; Dendritic Cells; Dimethyl Sulfoxide; Dose; Effector Cell; Electrolytes; Ensure; FDA approved; Freezing; Gases; Glucose; Goals; HIV; Histology; Human; Immune; Immune response; Immunologics; Immunotherapy; In Vitro; Intervention; Investigation; Kentucky; Laboratories; Learning; Leukapheresis; Licensing; Liquid substance; Lymphokine-Activated Killer Cells; Malignant Neoplasms; Measures; Mechanics; Methodology; Methods; Monitor; Natural Killer Cells; Nitrogen; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Plasmalyte A; Preparation; Procedures; Process; Property; Protocols documentation; Reagent; Rehydrations; Reliability of Results; Research Personnel; Running; Sampling; Serum; Serum-Free Culture Media; Site; Solutions; Source; T-Lymphocyte; Techniques; Temperature; Testing; Therapeutic; Time; Transplantation; Treatment Protocols; United States; Universities; Vaccines; Variant; Viral; Work; cell preparation; clinical research site; cost; crystalloid; in vivo; innovation; insight; melanoma; precursor cell; research clinical testing; research study; response; standard measure; time interval; tumor

DESCRIPTION (provided by applicant): There are many immunotherapy trials being conducted throughout the world ranging from vaccines to T cell transfer and antibody studies. In these trials, aside from the obvious need to assess clinical responses, Investigators must analyze changes in immune responsiveness which results from the immune intervention. Thus, there is a need to collect PBMC from patients at regular intervals both before and following the therapy. These PBMC are cryopreserved to save the PBMC phenotype and function until a later time point when the PBMC can be thawed and assessed to determine if the immunotherapy was effective. In addition, preparation of subsequent doses benefits from a source of readily obtainable PBMC which will respond to in vitro manipulation similar to the day the product was received by the lab. There are few standardized techniques available for cell cryopreservation as it relates to immunotherapy. Most labs have their own protocols which themselves often provide inconsistencies in cell viability and function upon thawing. It is also difficult to evaluate results of immunotherapy trials and make comparisons from lab to lab. The current proposal will focus on cryopreservation medium and compare static vs. controlled rate freezing techniques. In 4 Specific aims, we will examine: 1) optimum medium for cryopreservation. Human serum + DMSO will be compared to Plasmalyte-A, an FDA approved commercial serum free electroyte or rehydration fluid. The comparison of Plasmalyte A to serum is important because if it works, significant cost reduction will occur since the cost per liter is less than 1% of the cost of serum. In addition, this is an FDA approved product which is consistent from batch to batch. The second specific aim will evaluate different methods of thaw. Specific Aim 3 will compare static freeze techniques to controlled rate freezing. The fourth specific aim will monitor Specific aims 1 and 3 by comparing PBMC subset function at various time intervals using assays of cellular immunity. The goal of the proposal will be to develop a strategy for peripheral blood mononuclear cell (PBMC) cryopreservation which can be more universally applied. A more standardized approach will allow a more accurate assessment of immne responsivenss which can be compared between studies in different centers. In addition, other clinical studies utilizing PBMC can also benefit from such a study (studies of HIV and transplantation for instance). The 4 specific aims will be done over a two year period of time.

描述（由申请人提供）：世界各地正在进行许多免疫治疗试验，从疫苗到 T 细胞转移和抗体研究。在这些试验中，除了明显需要评估临床反应外，研究人员还必须分析免疫干预导致的免疫反应性变化。因此，需要在治疗之前和之后定期从患者收集 PBMC。这些 PBMC 被冷冻保存以保存 PBMC 表型和功能，直到稍后的时间点可以解冻和评估 PBMC 以确定免疫疗法是否有效。此外，后续剂量的制备受益于易于获得的 PBMC 来源，该来源将对类似于实验室收到产品当天的体外操作做出反应。由于与免疫治疗相关，因此很少有标准化技术可用于细胞冷冻保存。大多数实验室都有自己的方案，这些方案本身通常会导致解冻时细胞活力和功能的不一致。评估免疫治疗试验的结果并在实验室之间进行比较也很困难。目前的提案将重点关注冷冻保存介质，并比较静态与控速冷冻技术。在 4 个具体目标中，我们将研究：1) 冷冻保存的最佳培养基。人血清 + DMSO 将与 Plasmalyte-A 进行比较，Plasmalyte-A 是 FDA 批准的商业无血清电解质或补液。 Plasmalyte A 与血清的比较很重要，因为如果它有效，将会显着降低成本，因为每升的成本不到血清成本的 1%。此外，这是 FDA 批准的产品，批次之间是一致的。第二个具体目标将评估不同的解冻方法。具体目标 3 将比较静态冷冻技术与控速冷冻技术。第四个具体目标将通过使用细胞免疫测定比较不同时间间隔的 PBMC 子集功能来监测具体目标 1 和 3。该提案的目标是制定一种可以更普遍应用的外周血单核细胞（PBMC）冷冻保存策略。更标准化的方法将能够更准确地评估免疫反应，并可以在不同中心的研究之间进行比较。此外，利用 PBMC 的其他临床研究也可以从此类研究中受益（例如 HIV 和移植的研究）。这4个具体目标将在两年内完成。