Preclinical Testing of Interleukin-7 in Primates

Interleukin-7 在灵长类动物中的临床前测试

• 批准号: 6644818
• 负责人: JAN STOREK
• 金额: $ 44.27万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-15 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6644818
• 关键词: B lymphocyte; T cell receptor; T lymphocyte; autoimmune disorder; autologous transplantation; baboons; blood cell count; bone density; computed axial tomography; flow cytometry; granulocytopenia; green fluorescent proteins; helper T lymphocyte; hematopoietic stem cells; immunopathology chemotherapy; interleukin 7; laboratory mouse; lymphoma; lymphopoiesis; nonhuman therapy evaluation; osteoporosis; placebos; polymerase chain reaction; stem cell transplantation; thymus

Background: Infections are frequent after high dose chemotherapy or total body irradiation followed by hematopoietic cell transplantation (HCT). This is due at least in part to the slow reconstitution of CD4 cells. In adult patients, the T cell repertoire is restricted and counts of phenotypically nave T cells are low for years. This is primarily due to the limited ability of adult patients to regenerate T cells de novo (from hematopoietic stem cells). Hypothesis: Interleukin-7 (IL7) hastens the regeneration of T cells de novo. Research design and Methods: Autologous HCT transplantation of irradiated baboons will be performed, using yellow fluorescent protein (YFP) gene-transduced CD34 cells and green fluorescent protein (GFP) gene-transduced T cells. After transplant, de novo T cell generation in IL7-treated versus placebo-treated animals will be evaluated by 1. enumerating YFP+ T cells, 2. determining functional T cell repertoire through the detection of various antigen-specific T cells after immunization with multiple antigens, 3. enumerating phenotypically nave (e.g., CD45RA high) CD4 T cells, 4. assessing the size and histology of the thymus, including RAG and TdT immunostains, thymocyte subsets and T cell receptor rearrangement excision circle (TREC) levels in thymocytes, and 5. determining TREC levels in peripheral T cells. To rule out the possibility that the expected result of increased YFP+/CD45RA high/TREC+ T cell counts in the blood of the IL7-treated animals is due to IL7-induced shift of stem cell-derived T cells from organs to the blood, YFP+, CD45RA high and TREC+ cells will be enumerated in both the blood and lymphatic tissues. We will also evaluate whether IL7 stimulates peripheral expansion of primate T cells by enumerating GFP+ T cells. Moreover, we will determine the efficacious (de novo regeneration-stimulating) dose of IL7 in primates and potentially toxicity including osteoporosis or a lymphoproliferative disorder. These results will be used for the design of clinical studies.

背景：造血细胞移植（HCT）后大剂量化疗或全身照射后经常发生感染。 这至少部分是由于CD 4细胞的缓慢重建。 在成人患者中，T细胞库受到限制，表型计数不明显，ve T细胞多年来一直很低。这主要是由于成年患者从头再生T细胞（从造血干细胞）的能力有限。假设：白细胞介素-7（IL-7）加速T细胞从头再生。研究设计和方法：将使用黄色荧光蛋白（YFP）基因转导的CD 34细胞和绿色荧光蛋白（GFP）基因转导的T细胞对辐照狒狒进行自体HCT移植。移植后，将通过1.计数YFP+ T细胞，2.通过在用多种抗原免疫后检测各种抗原特异性T细胞来确定功能性T细胞库，3.计数表型NAve（例如，CD 45 RA高）CD 4 T细胞，4.评估胸腺的大小和组织学，包括RAG和TdT免疫染色、胸腺细胞亚群和胸腺细胞中的T细胞受体重排切除环（TREC）水平，和5.测定外周T细胞中的TREC水平。 为了排除IL 7处理动物血液中YFP+/CD 45 RA高/TREC+ T细胞计数增加的预期结果是由于IL 7诱导的干细胞来源的T细胞从器官向血液的转移的可能性，将在血液和淋巴组织中计数YFP+、CD 45 RA高和TREC+细胞。我们还将通过计数GFP+ T细胞来评估IL 7是否刺激灵长类T细胞的外周扩增。此外，我们将确定灵长类动物中IL 7的有效（从头再生刺激）剂量和潜在毒性，包括骨质疏松症或淋巴增生性疾病。 这些结果将用于临床研究的设计。