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目的探讨流式细胞分选技术在筛选HIV-1抗原特异性单个B细胞中的应用。方法通过HIV-1特异性探针结合多色流式细胞技术,标记HIV-1抗原特异性B细胞;并结合单细胞流式分选,从两例HIV-1慢性感染者体内分离抗原特异性单个B细胞。结果两个样本均有较强的中和活性,可以用作筛选中和抗体。两例感染者经流式分选后分别获得探针特异性目的细胞CD3-、CD8-、INDO-1 VIOLET-、CD14-、CD19+、CD20+、Ig M-、Ig G+、JRFLgp140+、JRFLgp140-D368R-各33和20个。结论通过流式分选技术结合HIV-1特异性探针可以获得单个HIV-1抗原特异性B细胞,为后续获得有广谱中和HIV-1能力的抗体,设计抗HIV-1药物和HIV-1疫苗免疫原提供技术支持。
Objective To investigate the application of flow cytometry in the screening of HIV-1 antigen-specific single B cells. Methods HIV-1 antigen-specific B cells were labeled by HIV-1-specific probe combined with multi-color flow cytometry. Antigen specificities were separated from two chronic HIV-1 infected patients by single cell flow cytometry Single B cells. Results Both samples had strong neutralizing activity and could be used as screening neutralizing antibodies. Two infected patients were obtained by flow cytometry probe specific target cells CD3-, CD8-, INDO-1 VIOLET-, CD14-, CD19 +, CD20 +, Ig M-, Ig G +, JRFLgp140 +, JRFLgp140-D368R- 33 and 20 each. Conclusion Single HIV-1 antigen-specific B cells can be obtained by flow cytometry combined with HIV-1-specific probes. For the purpose of obtaining antibodies capable of neutralizing HIV-1 in the future, anti-HIV-1 drugs and HIV -1 vaccine immunogen to provide technical support.