A TEST FOR HTLV-1 MEDIATED IN VIVO T-CELL EXPANSION

HTLV-1 介导体内 T 细胞扩增的测试

• 批准号: 6209872
• 负责人: MARK ALLEGRETTA
• 金额: $ 12.19万
• 依托单位: BIOMOSAICS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-15 至 2001-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6209872
• 关键词: T cell receptor; T lymphocyte; cell cycle; clinical research; communicable disease diagnosis; cytodiagnosis; diagnosis design /evaluation; gene mutation; human T cell leukemia; human T cell lymphotropic virus type 1; human subject; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; reporter genes; southern blotting; virus integration

The primary objective of this project is to determine if a surrogate selection approach utilizing mutation at a reporter gene (hypoxanthine- guanine phosphoribosyltransferase (hprt]) as a probe for in vivo cell division can be used as a diagnostic tool for detection of sub-clinical clonal T-cell expansion in human T lymphotropic HTLV-1 carriers. Such a test would be to allow preventive therapy (initially in the setting of HTLV-1-mediated adult T-cell leukemia) to be initiated before the expanding clone has progressed to a more aggressive and untreatable state. The objective will be achieved by testing blood samples from HTLV-1-infected individuals with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Wild type and hprt mutant T-cell clones will be isolated, and clonal identity determined by multiplex PCR and DNA sequencing of T-cell receptor (TCR) variable region beta chain and third complimentarity determining regions. In vivo- expanded clones will be identified, and representatives of the amplified clones will be tested for monoclonal HTLV integration as compared to that observed in the peripheral blood sample. These studies may provide new insights into the precise mechanism of HTLV-1 leukemogensis, and direct the development of more effective therapies. PROPOSED COMMERCIAL APPLICATIONS: Development of an automated test for clonal T-cell expansion would aid in the diagnosis and treatment of virally-mediated pathologies. The test would also have diagnostic/therametric applications in disease conditions where therapeutic agents are used that target the puring salvage pathway.

本项目的主要目的是确定利用报告基因（次黄嘌呤-鸟嘌呤磷酸核糖转移酶（hprt））突变作为体内细胞分裂探针的替代选择方法是否可用作检测人T淋巴细胞嗜性HTLV-1携带者亚临床克隆T细胞扩增的诊断工具。这样的测试将允许预防性治疗（最初在HTLV-1介导的成人T细胞白血病的情况下）在扩增克隆发展到更具侵略性和不可治疗的状态之前开始。将通过检测HTLV-1相关脊髓病/热带痉挛性下肢轻瘫（HAM/TSP）HTLV-1感染个体的血液样本来实现该目标。将分离野生型和hprt突变T细胞克隆，并通过T细胞受体（TCR）可变区β链和第三互补决定区的多重PCR和DNA测序确定克隆同一性。将鉴定体内扩增的克隆，并将检测扩增克隆的代表性，与外周血样品中观察到的单克隆HTLV整合进行比较。这些研究可能为HTLV-1白血病发生的确切机制提供新的见解，并指导更有效的治疗方法的开发。拟议的商业应用：开发克隆T细胞扩增的自动化测试将有助于诊断和治疗病毒介导的病理。该测试还将在使用靶向纯化挽救途径的治疗剂的疾病状况中具有诊断/治疗应用。