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PKH67 Green Fluorescent Cell Linker Midi Kit for General Cell Membrane Labeling


Company: Sigma-Aldrich
Catalog#: MIDI67
Other protocol()

ADCC Assay Protocol
[Abstract]  Antibody-dependent cell-mediated cytotoxicity (ADCC) bridges innate and adaptive immunity, and it involves both humoral and cellular immune responses. ADCC has been found to be a main route of immune protection against viral infections and cancers in vivo. Here we developed a flow cytometry based protocol for ADCC assay using human peripheral blood mononuclear cells (PBMCs) as effector cells. Using this protocol, we determined the ADCC activity of convalescent plasma IgGs from six H1N1-infected human subjects in China, and identified two dominant ADCC epitopes, designated E1 [amino acid (AA) 92-117] and E2 (AA 124-159), on haemagglutinin of pandemic H1N1 influenza virus by epitope mapping of the convalescent plasma IgGs with different levels of ADCC activity. Our study may aid in ... [摘要]  抗体依赖性细胞介导的细胞毒性(ADCC)桥接先天和适应性免疫,并且其涉及体液和细胞免疫应答。已经发现ADCC是针对体内病毒感染和癌症的免疫保护的主要途径。在这里,我们开发了基于流式细胞术的协议,使用人类外周血单核细胞(PBMCs)作为效应细胞ADCC测定。使用这个协议,我们确定在中国六个H1N1感染人类受试者的恢复期血浆IgG的ADCC活性,并确定两个显性ADCC表位,指定E1 [氨基酸(AA)92-117]和E2(AA 124-159) ,通过具有不同水平的ADCC活性的恢复血浆IgG的表位作图对大流行性H1N1流感病毒的血凝素进行。我们的研究可能有助于设计可以引发具有高ADCC活性的抗体的免疫原。设计为包括有效的广泛中和抗体和ADCC表位的结构决定簇的疫苗免疫原可以赋予针对病毒感染的全面免疫保护。