Prado Ilia, Fouts Timothy R, Dimitrov Antony S
Profectus BioSciences, Inc., Baltimore, MD, USA.
Methods Mol Biol. 2009;525:517-31, xiv. doi: 10.1007/978-1-59745-554-1_28.
Antibodies can neutralize HIV-1 with potency and cross-reactivity that varies widely and is related but not correlated to their antigen-binding affinity. Therefore, in addition to measuring binding affinity, an evaluation of the antibody neutralizing activity in tissue cultures is important for development of antibody-based therapeutics, design of candidate vaccine immunogens, and understanding the mechanisms of virus entry, neutralization, and evasion of immune responses. The development of a standardized assay for measurement of the in vitro neutralizing activities of the antibody has remained a challenging goal in the last two decades. There are two types of widely used assays, which vary in details between different laboratories--assays based on cell line/pseudovirus and assays based on infection of peripheral blood mononuclear cells (PBMCs). Here we describe in detail the PBMC-based assay, which is more laborious but in our opinion represents a closer approximation of the in vivo situation. As with all other in vitro assays the results of such measurements are only an indication of the antibody potency in vivo, and animal studies and ultimately clinical trials are needed for the development of such antibodies as potential prophylactics and therapeutics.
抗体能够中和HIV-1，其效力和交叉反应性差异很大，且与它们的抗原结合亲和力相关但不具有相关性。因此，除了测量结合亲和力外，评估抗体在组织培养中的中和活性对于基于抗体的治疗药物开发、候选疫苗免疫原设计以及理解病毒进入、中和和免疫逃逸机制都很重要。在过去二十年中，开发一种标准化的测定抗体体外中和活性的方法一直是一个具有挑战性的目标。有两种广泛使用的测定方法，不同实验室之间在细节上有所不同——基于细胞系/假病毒的测定方法和基于外周血单个核细胞（PBMC）感染的测定方法。在这里，我们详细描述基于PBMC的测定方法，该方法更费力，但在我们看来更接近体内情况。与所有其他体外测定方法一样，此类测量结果仅表明抗体在体内的效力，开发此类抗体作为潜在的预防和治疗药物需要进行动物研究并最终进行临床试验。
