Flow Cytometric Evaluation of Treatment Responses in Sickle Cell Disease

镰状细胞病治疗反应的流式细胞术评估

• 批准号: 7593137
• 负责人: irina maric
• 金额: $ 1.1万
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593137
• 关键词: Adult; Aging; American; Biological Assay; Cells; Child; Chronic; Collaborations; Dose; Edetic Acid; Erythrocytes; Erythropoietin; Evaluable Disease; Evaluation; Fetal Hemoglobin; Hereditary Disease; Incubated; Kidney Failure; Methods; Monitor; Monoclonal Antibodies; Morbidity - disease rate; Newborn Infant; Patients; Population; Procedures; Production; Reticulocytes; Risk; Sickle Cell; Sickle Cell Anemia; Solutions; Staining method; Stains; Syndrome; Techniques; Treatment Protocols; hydroxyurea; mortality; peripheral blood; response; sickling; thiazole orange

With the emergence of fetal hemoglobin (HbF)-stimulating agents as potential treatments for sickle-cell disease, procedures to monitor the effect of these agents on HbF levels in patients are needed. Flow cytometric assay was developed as a rapid procedure to detect fetal hemoglobin in erythrocytes (F cells) and reticulocytes (F reticulocytes) using a Tricolor-conjugated monoclonal antibody against HbF. In order to detect fetal hemoglobin in reticulocytes, these same erythrocytes are subsequently incubated with a staining solution of Thiazole Orange. This method permits rapid distinction and semi-quantitation of F cells and F reticulocytes in EDTA anticoagulated peripheral blood. In collaboration with Dr. Mark Gladwin group, we are following patients with sickle syndromes that are treated with hydroxyurea (HU) and/or erythropoietin or darbepoietin (both referred to as EPO). Flow cytometric analysis is used to establish baseline levels of HbF and F reticulocytes in SCD patients and to assess their responses to different treatment regimens. The percent of HbF and F-cells increase in all evaluable patients is recorded and HU dosing in conjunction with EPO therapy is explored, particularly in high-risk SCD patients and in the setting of renal insufficiency, common to the aging sickle cell population.

随着胎儿血红蛋白（HBF）刺激剂的出现，作为镰状细胞疾病的潜在治疗方法，需要监测这些药物对患者HBF水平的影响的程序。 流式细胞仪测定是作为一种快速的程序，用于检测红细胞（F细胞）和网状细胞（F网状细胞）中使用三色偶联的单克隆抗体对HBF的胎儿血红蛋白。 为了检测网状细胞中的胎儿血红蛋白，随后将这些相同的红细胞与噻唑橙的染色溶液一起孵育。 这种方法允许在EDTA抗凝外周血中快速区分F细胞和F网状细胞。与Mark Gladwin Group博士合作，我们正在关注用羟基脲（HU）和/或促红细胞增生素或Darbepoietin治疗的镰状综合征患者（均称为EPO）。 流式细胞仪分析用于在SCD患者中建立HBF和F网状细胞的基线水平，并评估其对不同治疗方案的反应。记录所有可评估患者的HBF和F细胞的百分比增加，并探索HU剂量与EPO疗法结合使用，尤其是在高风险的SCD患者中以及在肾脏不足的情况下，这是老龄化镰状细胞种群常见的情况。