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: 7372208
• 负责人: JOHN R. YANNELLI
• 金额: $ 17.58万
• 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 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7372208
• 关键词: Acute; Address; Aftercare; Antibodies; Area; Biological Assay; Blood; Boxing; Cell Survival; Cell physiology; Cells; Cellular Assay; Cellular Immunity; Clinical; Clinical Research; Clinical Trials; Condition; Cryopreservation; Dendritic Cells; Dimethyl Sulfoxide; Dose; Effector Cell; Electrolytes; Ensure; Freezing; Gases; Glucose; Goals; Guanosine Monophosphate; 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; Purpose; Range; Rate; Reagent; Rehydrations; Reliability of Results; Research Personnel; Running; Sampling; Serum; Serum-Free Culture Media; Site; Solutions; Source; Standards of Weights and Measures; T-Lymphocyte; Techniques; Temperature; Testing; Therapeutic; Time; Transplantation; Treatment Protocols; United States; United States Food and Drug Administration; Universities; Vaccines; Variant; Viral; Work; cell preparation; clinical research site; cost; crystalloid; day; human study; in vivo; innovation; insight; melanoma; precursor cell; research clinical testing; research study; response; 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）：全世界正在进行许多免疫疗法试验，从疫苗到T细胞转移和抗体研究。在这些试验中，除了显然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，需要在治疗前后定期从患者那里收集PBMC。这些PBMC是冷冻保存的，以节省PBMC表型和功能，直到可以解冻和评估PBMC的时间点，以确定免疫疗法是否有效。此外，随后的剂量的制备益处从可容易获得的PBMC来源中受益，这将响应与实验室收到产品的那一天相似的体外操作。与免疫疗法有关的细胞冷冻保存几乎没有标准化技术。大多数实验室都有自己的协议，这些方案本身通常会在细胞活力和解冻时功能上提供不一致的情况。也很难评估免疫疗法试验的结果并与实验室进行比较。当前的建议将集中于冷冻保存介质，并比较静态速率与受控速率冻结技术。在4个特定目标中，我们将研究：1）冷冻保存的最佳培养基。将将人血清 + DMSO与FDA认可的商业血清无元素或再合化液的质子分析进行比较。血浆A与血清的比较很重要，因为如果起作用，每升成本少于血清成本的1％，则会大幅降低成本。此外，这是FDA批准的产品，从批次到批量是一致的。第二个特定目的将评估不同的融化方法。特定的目标3将将静态冻结技术与控制速率冻结进行比较。第四个特定目标将使用细胞免疫测定在各种时间间隔比较PBMC子集功能，从而监测特定目标1和3。该提案的目的是制定外周血单核细胞（PBMC）冷冻保存策略，该策略可以更普遍地应用。一种更标准化的方法将允许对IMMNE球的评估进行更准确的评估，这些评估可以在不同中心的研究之间进行比较。此外，其他利用PBMC的临床研究也可以从此类研究中受益（例如，HIV和移植研究）。这4个具体目标将在两年的时间内完成。