GUIDELINE FOR FLOW CYTOMETRIC IMMUNOPHENOTYPING - A REPORT FROM THE NATIONAL-INSTITUTE-OF-ALLERGY-AND-INFECTIOUS-DISEASES, DIVISION OF AIDS
GUIDELINE FOR FLOW CYTOMETRIC IMMUNOPHENOTYPING - A REPORT FROM THE NATIONAL-INSTITUTE-OF-ALLERGY-AND-INFECTIOUS-DISEASES, DIVISION OF AIDS
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DOI：
10.1002/cyto.990140703
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发表时间：
1993-10-01
期刊：
影响因子：
--
通讯作者：
KAGAN, J
中科院分区：
文献类型：
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作者：
CALVELLI, T;DENNY, TN;KAGAN, J
CD4阳性T淋巴细胞的进行性和选择性耗竭是HIV感染和艾滋病的一个特征。在HIV感染者中，CD4阳性T细胞的减少与机会性感染和疾病进展的风险增加有关。在HIV感染中，CD4T细胞计数是临床预后、治疗监测和临床试验进入标准最常用的实验室指标。目前，CD4T细胞的计数包括三个不同的测量：白细胞计数(WBC)、淋巴细胞分化(淋巴细胞百分比)和CD4阳性T淋巴细胞的百分比(CD4%)。使用自动血液学仪器对全血样本进行常规的WBC和淋巴细胞百分比测量。采用流式细胞仪免疫分型法检测全血中的CD4百分率。样本与针对细胞表面抗原(例如，CD4)的氟铬标记的单抗反应。当在流式细胞仪上进行分析时，结合了标记抗体的细胞可以根据它们的荧光发射进行量化。同时，使用光散射特性根据大小和粒度将淋巴细胞从其他白细胞(粒细胞、单核细胞)中分离出来。由此得到的CD4%用于通过以下公式计算CD4细胞数(即绝对的CD4、CD4计数)：
Progressive and selective depletion of CD4-positive T lymphocytes is a hallmark of HIV infection and AIDS. Decreases in CD4-positive T cells are associated with increased risk for opportunistic infection and disease progression in HIV-infected individuals. In HIV infection, the CD4 T-cell count is the most commonly utilized laboratory measure for clinical prognosis, therapeutic monitoring, and entry criteria for clinical trials. Currently, the enumeration of CD4 T cells involves three distinct measurements: the white blood count (WBC), the lymphocyte differential (percent lymphocytes), and the percentage of T lymphocytes that are CD4 positive (CD4 percent). The WBC and percent lymphocytes are routinely measured on whole blood samples using an automated hematology instrument. The CD4 percent is measured on whole blood by flow cytometric immunophenotyping. The sample is reacted with fluorochromelabeled monoclonal antibodies that are specific for cellsurface antigens (eg, CD4). When analyzed on a flow cytometer, cells that have bound the labeled antibody can be quantified on the basis of their fluorescence emission. Simultaneously, lymphocytes are resolved from other white blood cells (granulocytes, monocytes) based on size and granularity using light scatter properties. The resulting CD4% is used to calculate the CD4 cell number (ie, absolute CD4, CD4 count) by the formula: