CYTOMETRIC CD4 COUNTING - A MINIMALIST APPROACH

细胞计数 CD4 计数 - 极简方法

• 批准号: 7338991
• 负责人: HOWARD M. SHAPIRO
• 金额: $ 9.78万
• 依托单位: HOWARD M. SHAPIRO, MD, P.C.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7338991
• 关键词: Acquired Immunodeficiency Syndrome; Adult; Affect; Africa; Antibodies; Area; Asia; CD4 Lymphocyte Count; CD4 Positive T Lymphocytes; Caribbean natives; Caribbean region; Cells; Charge; Child; Compatible; Complex; Computers; Consumption; Count; Country; Devices; Diagnostic; Disease; Eastern Europe; Flow Cytometry; Fluorescence; Foundations; Gold; Government; HIV; HIV-2; Homosexuals; Image Cytometry; Immunologic Deficiency Syndromes; Investigation; Journals; Label; Life; Lymphocyte Subset; Measurement; Medicine; Monitor; Monoclonal Antibodies; New England; Nongovernmental Organizations; Other Resources; Patients; Principal Investigator; Process; Public Health; Reagent; Research Infrastructure; Resolution; Resources; Retroviridae; Sampling; Semiconductors; South America; Standards of Weights and Measures; Syndrome; T-Lymphocyte; Technology; Testing; Training; United Nations; United States; antiretroviral therapy; base; cost; digital; fluorescence imaging; instrument; men; metal oxide; peripheral blood; prognostic; size; virtual

DESCRIPTION (provided by applicant): In late 1981, Gottlieb et al described an apparently new immunodeficiency syndrome in homosexual men living in the United States. We now know the syndrome as AIDS (Acquired Immune Deficiency Syndrome), recognize the retrovirus HIV (Human Immunodeficiency Virus) as its cause, and describe the involved cell subset as CD4-positive (CD4+) T cells. The absolute count of CD4+ T cells in peripheral blood was quickly identified as a valuable prognostic measurement in HIV-infected patients. AIDS originated in Africa, and it is there, and in other resource-poor areas of the world, that the disease has had the greatest impact on public health. Since effective antiretroviral therapy (ART) became available in the developed world in the 1990s, the United Nations, various governments, and nongovernmental organizations such as the Gates and Clinton Foundations, have sought to provide affordable treatment to at least some of the millions of HIV-infected adults and children in resource-poor countries in Africa, Asia, the Caribbean, Eastern Europe, and South America. Optimal planning and monitoring of such therapy would be facilitated if equally affordable CD4 counting technology were available. Flow cytometry is recognized as the "gold standard" for CD4 counting in affluent countries, in which several million tests are performed annually at a cost of tens of US dollars each. In resource-poor areas, an "affordable" CD4 counting test means that test must cost no more than a few dollars, and be usable in areas without the infrastructure typically required to support complex diagnostic apparatus such as flow cytometers. The measurement capabilities required to do CD4 counting can now be incorporated into substantially smaller, simpler, more energy- efficient, less expensive devices without any loss of accuracy and precision. This project will determine the minimal instrument configuration for a two-parameter low-resolution fluorescence imaging cytometer using light-emitting diodes (LEDs) for fluorescence excitation and a CCD (charge-coupled device) or CMOS (complementary metal oxide semiconductor) camera chip(s) for photodetection, suitable for CD4 counting using fluorescently labeled monoclonal antibodies, and requiring only minimal sample handling and operator training. It will also be determined whether combinations of antibodies bearing a single fluorescent label can be used to accurately identify CD4+ T cells and other lymphocyte subsets in a single-parameter fluorescence imaging or flow cytometer. AIDS (Acquired Immune Deficiency Syndrome) affects millions of adults and children worldwide, and it is in resource-poor areas of the world that the disease has had the greatest impact on public health. Various government and nongovernmental agencies have sought to provide affordable antiretroviral therapy (ART) to some of the millions of HIV-infected adults and children. Optimal planning and monitoring of therapy would be greatly facilitated by an affordable CD4 counting technology. This project will attempt to produce a simple, rugged, extremely inexpensive automated CD4 counting device using light-emitting diodes and digital camera and computer technology, and requiring only minimal sample handling and operator training.

描述（由申请人提供）：1981 年末，Gottlieb 等人描述了生活在美国的同性恋男性中一种明显新的免疫缺陷综合症。我们现在知道这种综合症为艾滋病（获得性免疫缺陷综合症），并认识到逆转录病毒 HIV（人类免疫缺陷病毒）是其病因，并将所涉及的细胞亚群描述为 CD4 阳性 (CD4+) T 细胞。外周血中 CD4+ T 细胞的绝对计数很快被确定为 HIV 感染患者的一个有价值的预后指标。艾滋病起源于非洲，正是在那里，以及世界上其他资源匮乏的地区，这种疾病对公众健康的影响最大。自 20 世纪 90 年代发达国家获得有效的抗逆转录病毒疗法 (ART) 以来，联合国、各国政府以及盖茨基金会和克林顿基金会等非政府组织一直致力于为数百万艾滋病毒感染者中的至少一部分提供负担得起的治疗-非洲、亚洲、加勒比地区、东欧和南美洲资源匮乏国家的受感染成人和儿童。如果有同样负担得起的 CD4 计数技术，将有助于对此类治疗进行最佳规划和监测。流式细胞术被公认为富裕国家CD4计数的“黄金标准”，每年进行数百万次检测，每次检测费用为数十美元。在资源匮乏的地区，“负担得起”的 CD4 计数测试意味着测试成本必须不超过几美元，并且可以在没有支持复杂诊断设备（例如流式细胞仪）通常所需的基础设施的地区使用。 CD4 计数所需的测量功能现在可以集成到更小、更简单、更节能、更便宜的设备中，而不会损失任何准确性和精度。该项目将确定使用发光二极管 (LED) 进行荧光激发和 CCD（电荷耦合器件）或 CMOS（互补金属氧化物半导体）相机芯片的双参数低分辨率荧光成像细胞仪的最小仪器配置。 s) 用于光电检测，适用于使用荧光标记的单克隆抗体进行 CD4 计数，并且仅需要最少的样品处理和操作员培训。还将确定带有单一荧光标记的抗体组合是否可用于在单参数荧光成像或流式细胞仪中准确识别 CD4+ T 细胞和其他淋巴细胞亚群。艾滋病（获得性免疫缺陷综合症）影响着全世界数以百万计的成年人和儿童，而在世界资源匮乏的地区，这种疾病对公共卫生的影响最大。各个政府和非政府机构一直在寻求为数百万感染艾滋病毒的成人和儿童中的一些人提供负担得起的抗逆转录病毒治疗（ART）。经济实惠的 CD4 计数技术将极大地促进治疗的最佳规划和监测。该项目将尝试使用发光二极管、数码相机和计算机技术生产一种简单、坚固、极其便宜的自动 CD4 计数装置，并且只需要最少的样品处理和操作员培训。