Rodríguez-Sáinz C, Valor L, Hernández D C, Gil J, Carbone J, Pascual-Bernaldez M, Rodríguez-Alcántara F, Martínez I, Vicario J L, Mallal S, Fernández-Cruz E
Servicio de Inmunología Clínica, Hospital General Universitario Gregorio Marañón, Departamento de Microbiología I (Inmunología), Universidad Complutense de Madrid, Madrid, Spain.
HIV Clin Trials. 2013 Jul-Aug;14(4):160-4. doi: 10.1310/hct1404-160.
Rapid screening for the detection of HLA-B*57:01 in the prevention of abacavir hypersensitivity in HIV-1-infected patients is a hallmark for clinical services.
The aim of this work was to analyze the utility of flow cytometry with a new FITC-conjugated B-17 monoclonal antibody (mAb3E12) for HLA-B*57:01 screening in a Spanish cohort of 577 HIV-1+ individuals.
Cryopreserved peripheral blood mononuclear cell samples from HIV-1+ individuals were analyzed by flow cytometry with the mAb 3E12 that recognizes both HLA-B57 and HLA-B58 alleles (members of the group specificity, HLA-B17). Patients' DNA samples had been previously typed for HLA-B*57:01 with PCR-SSO or PCR-SSP and additional DNA sequencing (EPI Study). The results obtained by flow cytometry were compared with the results obtained by the DNA-PCR techniques.
By flow cytometry, 46 samples (7.97%) were positive for HLA-B17, 530 (91.86%) were negative, and 1 (0.17%) was undetermined. All samples found negative by flow cytometry were negative for HLA-B57:01 by DNA-PCR. Of the HLA-B17 positive samples, 31 (67.4%) were positive for HLA-B57:01, 2 (3.25%) were positive for HLA-B57:03, 11 (26.1%) were positive for HLA-B58, and 2 (3.25%) were negative for both HLA-B57 and HLA-B58 antigens. The undetermined sample was negative for HLA-B57 and HLA-B58 alleles by DNA-PCR.
This study shows that flow cytometry with mAb3E12 is a highly sensitive method (no false negatives) to implement prior to DNA-PCR analysis for rapid screening of HLA-B*57:01. Additional confirmation by molecular HLA typing method would be required in less than 10% of the cohort of HIV-1-infected individuals.
在预防HIV-1感染患者的阿巴卡韦超敏反应中，快速筛查HLA-B*57:01是临床服务的一项标志。
本研究旨在分析采用新型异硫氰酸荧光素（FITC）偶联的B-17单克隆抗体（mAb3E12）进行流式细胞术检测，对西班牙577名HIV-1阳性个体队列进行HLA-B*57:01筛查的实用性。
采用识别HLA-B57和HLA-B58等位基因（组特异性HLA-B17成员）的mAb 3E12，通过流式细胞术分析HIV-1阳性个体的冷冻保存外周血单个核细胞样本。患者的DNA样本先前已通过聚合酶链反应-序列特异性寡核苷酸探针杂交（PCR-SSO）或聚合酶链反应-序列特异性引物（PCR-SSP）以及额外的DNA测序进行HLA-B*57:01分型（EPI研究）。将流式细胞术获得的结果与DNA-PCR技术获得的结果进行比较。
通过流式细胞术检测，46份样本（7.97%）HLA-B17呈阳性，530份（91.86%）呈阴性，1份（0.17%）结果不确定。流式细胞术检测为阴性的所有样本，DNA-PCR检测HLA-B57:01均为阴性。在HLA-B17阳性样本中，31份（67.4%）HLA-B57:01呈阳性，2份（3.25%）HLA-B57:03呈阳性，11份（26.1%）HLA-B58呈阳性，2份（3.25%）HLA-B57和HLA-B58抗原均为阴性。结果不确定的样本DNA-PCR检测HLA-B57和HLA-B58等位基因为阴性。
本研究表明，在进行DNA-PCR分析之前，采用mAb3E12的流式细胞术是一种高度敏感的方法（无假阴性），可用于快速筛查HLA-B*57:01。在不到10%的HIV-1感染个体队列中，需要通过分子HLA分型方法进行额外确认。
